Vaccines against Chlamydia sp.

Abstract

The present invention describes an efficient vaccine against a Chlamydia trachomatis (Ct). The vaccine is based on recombinant fusion molecules that are capable of generating a high titered neutralizing antibody response that is protective against various Ct serovars. Our invention furthermore describe the combination of these antibody promoting fragments with Ct antigens that are targets for T cells with the aim to provide a vaccine that activate both arms of the immune system.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of the priority of U.S. Provisional Patent Application No. 61/802,907, filed Mar. 18, 2013, which application, including sequence listing, is incorporated by reference in its entirety. The benefit of the priority of Danish Patent Application Nos. PA 2013 00155, filed Mar. 18, 2013, and PA 2013 00684, Dec. 11, 2013, are also claimed, which applications, including sequence listings, are also incorporated herein by reference in their entireties.

FIELD OF INVENTION

[0002] The present invention relates to polypeptides of repetitive units of immunogenic fragments of surface exposed regions of outer membrane proteins of Chlamydia sp. and pharmaceutical compositions and vaccines comprising these fusion proteins.

BACKGROUND OF THE INVENTION

[0003] Chlamydiae are intracellular bacterial pathogens responsible for a variety of infections. Chlamydia pneumoniae is responsible for human acute respiratory infection and believed to play a role in coronary heart disease. Chlamydia trachomatis is the causative agent of human sexually transmitted disease and eye infections (Trachoma). Also in animals, several infections with Chlamydia sp. are known, e.g. Chlamydia Suis infecting pigs, and Chlamydiaphila abortus which causes abortion in small ruminants (sheep and goats).

[0004] Worldwide, it is estimated that 92 million individuals become sexually infected with Chlamydia trachomatis (Ct)¹. Urogenital infections with Ct are of public health concern because of its high prevalence and the fact that it's a risk factor for ectopic pregnancy and infertility². In addition to this Ct infections have been shown to facilitate the transmission of HIV³ and act as a co-factor in HPV-induced cervical carcinoma⁴. The duration of untreated genital Ct infection can be prolonged, and complete clearance is often not reached within the first 12 months⁵. From human studies it is known that some degree of protective immunity against genital re-infection develops, although it appears at best to be partial⁶. The infection is effectively controlled by antibiotic therapy; however the high prevalence of asymptomatic cases suggests that sustainable disease control can only be envisaged if an effective Chlamydia vaccine is developed.

[0005] A vaccine against Ct needs to elicit protective T-cell and B-cell immunity in the genital tract mucosa⁷. Immune mechanisms of clearance of infection and resistance to re-infection have been described in numerous studies. A variety of animal models and chlamydial species have been used in attempts to identify protective and damaging immune responses. A general consensus has emerged that, in mice, CD4+Th1 cell mediated immune responses plays a major role in the resolution of Ct infection⁸, 9, 10, whereas the role of humoral immunity in protection has remained less well defined. In guinea pigs immunity to chlamydial infection is mediated at least partly by secretory IgA at the mucosal surface¹¹, 12 and also in the mouse model there is increasing evidence to support a role for antibodies in protective immunity⁹. Data from animal models that has emerged over the last years clearly demonstrate that if antibodies are formed after the infection is established they play a minimal role, whereas their presence at the time of infection (e.g. in a secondary response) promotes significant levels of protection, an effect that is however clearly amplified in the presence of Chlamydia specific CD4.sup.+ cells 9, 13, 14. A strong cell mediated immune (CMI) response without antibodies may on the other hand control bacterial replication but can in the worst case exacerbate the pathology associated with Chlamydia infection¹⁵ 16. The importance of this interplay between cell mediated immunity and antibodies is also becoming increasingly clear to support a preferential role of neutralizing antibodies in the initial phase of infection, whereas CD4.sup.+ cells are the main effectors throughout the rest of the infection¹⁷ 18 19. In summary balancing the immune effector mechanisms between antibodies and T cells seems to be crucial for disease outcome. We and others have identified a range of chlamydial antigens recognized during a natural infection in either humans or animal models²⁰, 21 22, 23 24 25, 26 27. Especially the publishing of the genome sequence in 1998 and modern high throughput techniques have led to the testing of almost the entire genome of 875 open reading frames²⁵. Importantly, identifying proteins as antigenic during an infection do not necessarily mean they are protective as vaccines²⁹ and despite the characterization of such a large number of antigens only very few of these have been demonstrated to mediate protection as vaccines in animal models³⁰ 31, 32. Furthermore for the majority of the vaccines recently reported the partial protection observed is mediated by T cells with no neutralizing antibodies. Therefore there is a lack of vaccine candidates that generate neutralizing antibodies that can cope with the infection in the initial phase and creating a balanced immune response.

[0006] Until now there has only been convincing data on neutralizing antibodies with three surface exposed antigens; PorB, which localized in the chlamydial outer membrane and functions as a porin³³. Antibodies against this has been shown to neutralize chlamydial infectivity³⁴ patent ref: U.S. Pat. No. 7,105,171. Another more recent antigen is PmpD. This protein has been shown to generate neutralizing antibodies in vitro, however the in vivo relevance of these antibodies have not yet been demonstrated³⁵.

[0007] MOMP is the classical target antigen for neutralizing antibodies and one of the first antigenic molecules described. It is a surface-exposed trans membrane protein which has structural (porin) properties³⁶, 37, 38. MOMP is a 40 kDa protein making up roughly 60% of the protein in the Ct membrane and is a target for neutralizing antibodies with proven efficacy both in vitro and in vivo. MOMP consists of four variable surface exposed domains (VD-1 to VD-4) separated by five constant segments³⁶ 39 and it is the molecular basis of the serovar (˜15) grouping of Chlamydia (FIG. 1). The in vitro and in vivo neutralizing antibody epitopes have been mapped to these VDs⁴⁰ 41 42 43 as The distribution profile of Ct urogenital serovars has been described for regions worldwide, providing epidemiological data for the serovar coverage needed of a MOMP based vaccine. The most common serovar detected worldwide is E (22-49% of cases) followed by serovars F and D (17-22% and 9-19%, respectively)⁴⁵ 46 47 48 49 50, meaning that a vaccine targeting serovars E, D and F would have a significant impact and cover more than 70% of the human population.

[0008] MOMP is highly immunogenic in humans and animals and has therefore been studied in great detail as a vaccine candidate, both as a natively purified protein, recombinantly and as DNA-vaccine. These vaccination attempts gave variable results¹⁷, 51, 52, 53, 54, 55, 56, 57. The reason for the relative inconsistency of MOMP as a vaccine is not fully understood, but the fact that the synthetic MOMP immunogens do not mimic the native structure of the protein has been the major concern⁵⁴. In this regard, the structure of this membrane bound cysteine rich molecule and refolding various products to achieve native protein structure has been extremely challenging and is not suitable for large scale vaccine production⁵⁸. Therefore, although clearly with vaccine potential, full size MOMP has so far not been a feasible vaccine candidate and several attempts have therefore been made to construct a vaccine based on selected epitopes (such as the highly conserved TTLNPTIAG (SEQ ID NO 76) in VD4³⁶, 59) or based on selected regions rich in neutralizing target epitopes (such as the VD's) from MOMP (WO9406827, U.S. Pat. No. 6,384,206)⁶⁰, 61 62, 63 64 51, 65 66.

[0009] There has been special focus on VD1, VD2 and VD4 because neutralizing monoclonal antibodies used for serotyping has been shown to map to these regions. These VD regions are targeted by antibodies during natural infection and in line with this, these regions have naturally been the focus of attempts to develop immuno-diagnostics. For example Mygind et al. constructed different polyantigens containing VD regions from different serovariants in the search for a diagnostic tool based on ELISA⁶⁷. This analysis revealed that by increasing the number of serovariants and include the species specific TTLNPTIAG into one recombinant polyantigen, it was possible to increase the specificity and sensitivity of the assay compared to an assay based on a single serovariant antigen.

[0010] Mainly VD4 has attracted interest as an immunogen because this region was shown to contain the highly conserved species-specific epitope TTLNPTIAG embedded in the variable region. Importantly, this conserved epitope in the VD4 region can elicit a broadly cross-reactive immune response, which is able to neutralize multiple serovars, among them the most prevalent D, E and F (FIG. 2). Peptides representing the VD4 region or the conserved epitope derived from this region have been used for immunization either alone, as chimeric peptides fused to other regions such as VD1 or mixed with T cell epitopes to potentiate the antibody response⁶⁰, 68 51, 65 64 69. All these constructs generated antibodies with some functional capabilities of neutralizing the infection in vitro but in general these strategies suffer from a low immunogenicity and the titres did not translate into in vivo protective efficacy against genital chlamydial challenge.

[0011] Reasons for the lack of protection when using these peptide based constructs can be numerous; including route of administration, type of immune response elicited, challenge dose, but most likely reflects that the vaccine molecule is not sufficiently immunogenic for use as a vaccine. The VD4 based strategy furthermore suffers from the limitation that with the exception of the TTLNPTIAG (SEQ ID NO 76) epitope, these fragments as mentioned above are highly specific for one or two serovariants and a vaccine would accordingly have to be composed of several components to cover the most frequent serovariants causing human disease.

[0012] In WO2012172042 it has previously been disclosed that B-cell epitopes within the VD regions, combined with defined T cell (Th1 and Th2) epitopes from non-variable domains of MOMP, could function as a poly-epitope vaccine against Chlamydia psitattci serovar D in chickens; in the examples they describe the combination of up to three B-cell epitopes each derived from a VD region from different variable domains of the same serovariant together with several T-cell epitopes. The use of repeats of a variable domain of a surface exposed region of MOMP and using different serovariants is not suggested and thus high titers and a broad response against different serovariants is not obtained.

[0013] The object of the current invention is to prepare recombinant fusion molecules that are capable of generating a high titered neutralizing antibody response that is protective against various Ct serovars in vivo. Our invention furthermore describes the combination of these antibody promoting fragments with Ct antigens that are targets for T cells with the aim to provide a vaccine that activate both arms of the immune system.

SUMMARY OF THE INVENTION

[0014] The present invention discloses an efficient vaccine against a pathogen, e.g. Chlamydia trachomatis (Ct), that incorporates repeats of surface exposed fragments of Ct antigens (homologous immuno-repeats) for maximal antibody responses. In one embodiment of the invention, these surface exposed fragments are extended to cover the flanking region of the surface exposed fragments that may contain T cell epitopes. One example is a defined large fragment representing an extended version of the VD1 or VD4 region from the Ct MOMP antigen and in the immuno-repeat format provides high levels of surface binding and neutralizing antibodies against Ct. In another important embodiment the immuno-repeat technology is used to obtain high titers and a broad response against different serovariants by the fusion of fragments that contain variable B and T cell epitopes from different serovariants (heterologous immuno-repeats). In yet another embodiment of our invention these surface exposed repeats are recombinantly fused with fragments of other surface exposed antigens such as PMPs or OMPs. Finally our invention discloses combinations of these immuno-repeat constructs with strong T cell antigens, such as MOMP(CT681), CT043 or CT004 from Ct that together form a very efficient vaccine against the different infectious stages of Ct infection.

FIGURE LEGENDS

[0015] FIG. 1. Model of MOMP (Serovar D, strain: D/B-120) membrane topology adapted from Findlay et al⁷⁷. The VD1, VD2, VD3 and VD4 are marked by black lines in the AA sequence (SEQ ID NO: 68) and in the linear model MOMP depicted interspaced with 5 constant segments (CS).

[0016] FIG. 2. Alignment of amino acid sequence of Ct MOMP VD4^ext for serovars D, E, F, G, Ia and J. The serovar D sequence is used as prototype, and conserved amino acids in other serovars are shown as ".". The variable domain VD4 according to Baehr et al (PNAS, 1988)³⁶ is shaded in gray and the conserved epitope TTLNPTIAG (SEQ ID NO: 76) is boxed. SvD VD4^ext (SEQ ID NO: 23), SvE VD4^ext (SEQ ID NO: 24), SvF VD4^ext (SEQ ID NO: 25), SvG VD4^ext (SEQ ID NO: 26), SvI VD4^ext (SEQ ID NO: 27), and SvJ VD4^ext (SEQ ID NO: 26) are shown.

[0017] FIG. 3. Model of MOMP (Serovar D, strain: D/B-120) membrane topology adapted from Findlay et al. The VD1ext and VD4ext described in this invention are shown as shaded in the figure. Amino acid sequence shown is SEQ ID NO: 68.

[0018] FIG. 4. Illustration of the design of homologous and heterologous immuno-repeats. The immuno-repeats are fusion proteins of e.g. four VD4^ext regions, either from the same serovar, homologous immuno-repeats, or from different serovars, heterologous immuno-repeats. The variable VD4 region within each VD4^ext region is illustrated as hatched.

[0019] FIG. 5. Enhanced and broadened immune responses after immunization with homologous immuno-repeats of VD4^ext compared with a monomeric VD4^ext unit.

[0020] FIG. 6. A construct composed of heterologous immuno-repeats from SvD, E, F and G induced a stronger response to multiple serovars compared to homologous immuno-repeats from SvF.

[0021] FIGS. 7A-7D. Fine specificity of the antibody responses after immunization with a heterologous immuno-repeat of the extended VD4 units from SvD, E, and F (CTH89) compared to constructs composed of a homologous immuno-repeat from (SvE^extVD4)*4 and from (SvF^extVD4)*4. In FIGS. 7A-7B, the Serovar E sequence shown is SEQ ID NO: 24, and the Serovar F sequence shown is SEQ ID NO: 25. In FIGS. 7C and 7D, each set of overlapping peptides is NMFTPYIGV through MQIVSLQLN, corresponding to SEQ ID NO: 195 through SEQ ID NO: 254, respectively. (See Example 3).

[0022] FIG. 8. Immunization with heterologous immuno-repeats of extended VD4's from SvD, SvE and SvF (CTH89) generates early T cell independent protection after a SvD challenge.

[0023] FIG. 9. In vivo neutralization with CTH89 specific serum.

[0024] FIG. 10. Coupling of heterologous immuno-repeats to recombinant MOMP.

[0025] FIG. 11. Vaccination with heterologous immuno-repeats of VD1-VD4's regions from SvD, SvE and SvF (CTH88) compared to vaccination with a single VD1-VD4 unit from SvD (CTH87)

[0026] FIG. 12. Coupling of T cell antigens to immuno-repeats of VD4

[0027] FIG. 13. Immunization with a cocktail of a heterologous VD4 immuno-repeat and a T cell antigen fusion molecule

[0028] FIG. 14. Comparison of CAF01 and Alum as adjuvant delivery system.

[0029] FIG. 15. Vaccination with heterologous immuno-repeats composed of reduced length of the VD4^ext regions from SvD, SvE, SvF and SvG. The CTH518 sequence shown is amino acids 1 through 68 of SEQ ID NO 53. The CTH286 sequence shown is amino acids 21 through 64 of SEQ ID NO 53. The CTH285 sequence shown is amino acids 28 through 57 of SEQ ID NO: 53.

[0030] FIG. 16 Vaccination with heterologous immuno-repeats composed of extended VD4^ext regions from SvD, SvE, SvF, SvG, SvIa and SvJ. The CTH88 sequence shown is amino acids 60 through 127 of SEQ ID NO: 46. The CTH69+CTH72 sequence shown is SEQ ID NO: 255.

DETAILED DISCLOSURE OF THE INVENTION

[0031] The invention discloses a polypeptide comprising

a) an amino acid sequence comprising one or more surface exposed fragments of the same outer membrane protein expressed in a serotype of Chlamydia sp.; and b) two or more additional amino acid sequences which is either the same sequence as defined in a) or is the corresponding surface exposed fragments from a variant of said outer membrane protein expressed in a serotype of Chlamydia sp., which is different from the serotype in a).

[0032] The invention thus discloses polypeptides comprising immuno-repeats, which is 3 or more such as 4 or more repeats of an amino acid sequence comprising an immunogenic portion of a surface exposed region of an outer membrane protein of Chlamydia sp. Hence the invention can be described as a polypeptide comprising an amino acid sequence comprising one or more surface expose fragments of the same outer membrane protein expressed in a serotype of Chlamydia sp. and two or more such as three or more additional amino acid sequences which is either the same sequence as defined in a) or is the corresponding surface exposed fragments from a variant of said outer membrane protein expressed in a serotype of Chlamydia sp., which is different from the serotype in a).

[0033] In a preferred embodiment the polypeptide comprises 3 or more different amino acid sequences, where said amino acid sequences each comprises one or more surface exposed fragments from different variants or isotypes of the same outer membrane protein that varies in different Chlamydia sp. serotypes, said amino acid sequences derived from different Chlamydia sp. serotypes (heterologous immuno-repeats in our terminology), but the invention also discloses a polypeptide comprising 3 or more repetitions of an amino acid sequence, where said amino acid sequence comprises one or more surface exposed fragments of the same outer membrane protein that varies in different Chlamydia sp. serotypes, said amino acid sequences derived from the same Chlamydia sp. serotype (homologous immuno-repeats in our terminology).

[0034] The outer membrane protein is preferable the major outer membrane protein (MOMP) from any Chlamydia sp. serotype and the surface exposed fragment is chosen from variable domain 1 (VD1), variable domain 2 (VD2), variable domain 3 (VD3) or variable domain 4 (VD4) of MOMP. The surface exposed fragment can optionally be linearized by substitution of cysteine in the amino acid sequence to prevent disulfide bonds.

[0035] A preferred embodiment of the invention is polypeptides comprising immuno-repeats with 3 or more repeats of the variable domain 4 (VD4) of MOMP from any of serovars D, E, F, G, Ia and J of Chlamydia trachomatis, where each variable domain consists of an amino acid sequence, which corresponds to the position of amino acid residues Nos. 309-338 in the amino acid sequence of MOMP of Chlamydia trachomatis serovar D (SvD) (SEQ ID NO.: 68) and where the variable domains in the immune-repeat is independently selected from the group consisting of the VD4 of serovar D, the VD4 of serovar E, the VD4 of serovar F, the VD4 of serovar G, the VD4 of serovar Ia and the VD4 of serovar J of Chlamydia trachomatis or has 80% sequence identity herewith.

[0036] The amino acid sequence of VD4 from serovar D, E, F, G, Ia and J corresponds to SEQ ID NO 15-20 respectively. Each variable domain can additionally be flanked/extended on the N-terminal side by either

[0037] i) The amino acid sequence EWQASLALSYRLNMFTPYIGVKWSRASFDADTIRIAQPK (SEQ ID NO 21) or

[0038] ii) A subsequence of the amino acid sequence in i) said subsequence comprising 1 or more amino acid residues,

[0039] On the C-terminal side the variable domain can additionally be flanked/extended by

[0040] iii) The amino acid sequence DTMQIVSLQLNKMKSRKSCGIAVGTTIVDA (SEQ ID NO 22)

[0041] iv) A subsequence of the amino acid sequence in iv) said subsequence comprising 1 or more amino acid residues, or an amino acid sequence which has at least 80% sequence identity herewith.

[0042] Hence the preferred embodiment can be described as polypeptides comprising 2-8 different amino acid sequences each derived from MOMP from Chlamydia trachomatis which comprises an amino acid sequence defined in formula I:

xx₁-VD4-xx₂ (Formula I)

wherein VD4 is independently selected from SEQ ID NO. 15-20 or an amino acid sequence which has at least 80% sequence identity herewith, and xx₁ consists of i) The amino acid sequence EWQASLALSYRLNMFTPYIGVKWSRASFDADTIRIAQPK (SEQ ID NO 21) or ii) A subsequence of the amino acid sequence in i) said subsequence comprising 1-38 amino acid residues, starting with the C-terminal K in the amino acid sequence in i) and xx₂ consists of iii) The amino acid sequence DTMQIVSLQLNKMKSRKSCGIAVGTTIVDA (SEQ ID NO 22)

[0043] v) A subsequence of the amino acid sequence in iii) said subsequence comprising 1-29 amino acid residues, starting with the N-terminal D in the amino acid sequence in iii).

[0044] Examples of fusion proteins comprising immuno-repeats of VD4 of MOMP is indicated by SEQ ID NO 49-59.

[0045] In another embodiment of the invention the polypeptide additionally comprises immuno-repeats of 3 or more variable domain 1 (VD1) of MOMP from any of serovars D, E, F, G, Ia and J of Chlamydia trachomatis, each variable domain consisting of an amino acid sequence, which corresponds to position of amino acid residues nos. 91-105 in the amino acid sequence of MOMP of Chlamydia trachomatis serovar D (SvD) (SEQ ID NO 68) and is independently selected from the group consisting of the VD1 of serovar D, the VD1 of serovar E, the VD1 of serovar F, the VD1 of serovar G, the VD1 of serovar Ia and the VD1 of serovar J of Chlamydia trachomatis or has 80% sequence identity herewith.

[0046] The amino acid sequence of VD1 from serovar D, E, F, G, Ia and J corresponds to SEQ ID NO 1-6 respectively. Each variable domain can additionally be flanked/extended on the N-terminal side by either

[0047] vi) The amino acid sequence SMRVGYYGDFVFDRVLKTDVNKEFQMG (SEQ ID NO 77)

[0048] vii) A subsequence of the amino acid sequence in v) said subsequence comprising 1 or more amino acid residues.

[0049] On the C-terminal side the variable domain can additionally be flanked/extended by

[0050] viii) The amino acid sequence NPAYGRHMQDAEMFTNAACMALNIWD (SEQ ID NO 78)

[0051] ix) A subsequence of the amino acid sequence in x) said subsequence comprising 1 or more amino acid residues;

[0052] Or an amino acid sequence which has at least 80% sequence identity herewith.

[0053] Hence another preferred embodiment can be described as polypeptides comprising 2-8 different amino acid sequences each derived from MOMP from Chlamydia trachomatis which comprises an amino acid sequence defined in formula I and additionally comprising an amino acid sequence defined in formula II:

yy₁-VD1-yy₂ (Formula II)

wherein VD1 is independently selected from SEQ ID NO. 1-6 or an amino acid sequence which has at least 80% sequence identity herewith, and yy₁ consists of v) The amino acid sequence DAISMRVGYYGDFVFDRVLKTDVNKEFQMG (SEQ ID NO 7) or vi) A subsequence of the amino acid sequence in v) said subsequence comprising 1-30 amino acid residues, starting with the C-terminal G in the amino acid sequence in v) and yy₂ consists of vii) The amino acid sequence NPAYGRHMQDAEMFTNAA (SEQ ID NO 8) or viii) A subsequence of the amino acid sequence in vii) said subsequence comprising 1-18 amino acid residues, starting with the N-terminal N in the amino acid sequence in vii).

[0054] Examples of polypeptides comprising immuno-repeats of VD1 is indicated by SEQ ID NO 9-14 and 45-48.

[0055] Further embodiments of the invention comprises additionally comprises a fragment comprising the variable domains 2 (VD2) and/or variable domains 3 (VD3) of MOMP respectively comprising an amino acid sequence defined in formula III and/or formula IV:

zz₁-VD2-zz₂ (Formula III)

qq₁-VD3-qq₂ (Formula IV)

wherein VD2 is independently selected from SEQ ID NO. 29-34 or an amino acid sequence which has at least 80% sequence identity herewith, and zz₁ consists of ix) The amino acid sequence TLGATSGYLKGNSASFNLVGLFG (SEQ ID NO 35) or x) A subsequence of the amino acid sequence in ix) said subsequence comprising 1-23 amino acid residues, starting with the C-terminal G in the amino acid sequence in ix) and zz₂ consists of xi) The amino acid sequence WELYTDTTFAWSVGARAALWE (SEQ ID NO 36) or xii) A subsequence of the amino acid sequence in xi) said subsequence comprising 1-22 amino acid residues, starting with the N-terminal V in the amino acid sequence in xi).

[0056] And wherein

VD3 is independently selected from SEQ ID NO. 37-42 or an amino acid sequence which has at least 80% sequence identity herewith, and qq₁ consists of xiii) The amino acid sequence ATLGASFQYAQSKPKVEELNVLCNAAEFTINKPKGYVG (SEQ ID NO 43) or xiv) A subsequence of the amino acid sequence in xiii) said subsequence comprising 1-22 amino acid residues, starting with the C-terminal G in the amino acid sequence in xiii) and qq₂ consists of xv) The amino acid sequence TGTKDASIDYHEWQASLALSYRLNMFTPYIGVKWS (SEQ ID NO 44) or xvi) A subsequence of the amino acid sequence in xv) said subsequence comprising 1-35 amino acid residues, starting with the N-terminal T in the amino acid sequence in xv).

[0057] The immuno-repeats can be heterologous, that is where the variable domain is derived from different serotypes or they can be homologous, that is where the variable domain is derived one serotype. The preferred number of repeats are 2, 3, 4, 5, 6, 7 or 8 repeats.

[0058] Furthermore the immuno-repeats in the polypeptides can be linearized, that is cysteine residues are replaced with serine.

[0059] The polypeptides comprising immuno-repeats can additionally comprise a moiety that facilitate export of the polypeptide whens produced recombinantly (e.g. signal peptides), a moiety that facilitate purification of the polypeptide (e.g. his-tags) and/or a moiety which enhance the immunogenicity (e.g. a T cell antigen). The T-cell target can be chosen from a Ct antigen such as CT043, CT004, CT414, CT681 or part hereof. Examples of such fusion proteins are indicated by SEQ ID NO 60-67.

[0060] A polypeptide according to the invention having the following functional abilities:

a) neutralize C. trachomatis serovar D in vitro with a 50% neutralization titer of 10^-3 or less, when tested in an experimental set-up comprising the administering a heterologous immuno-repeats; b) neutralize C. trachomatis serovar D in vivo in at least 50% of the mice at day 7 post infection when tested in a mouse model comprising administering a heterologous immuno-repeats c) broaden the immune response to multiple serovars of C. trachomatis in vitro when administering heterologous immuno-repeats.

[0061] The present invention also discloses nucleic acids encoding above described polypeptides.

[0062] The disclosed polypeptides or nucleic acids are used for the preparation of a pharmaceutical composition such as a vaccine. The vaccine can additionally comprise a pharmacologically acceptable carrier (virus like particles), excipient, adjuvant (e.g. DDA/TDB or alum) or immune modulator. The pharmaceutical composition can be used for prophylactic or therapeutic use against Chlamydia sp. Infections, including infections with Chlamydia trachomatis or C. pneumoniae.

[0063] A method for preventing, treating and/or reducing the incidence of Chlamydia sp. Infections, including infections with Chlamydia trachomatis or C. pneumoniae, by administering this pharmaceutical composition is also disclosed.

[0064] In the following the invention will be described in more detail and exemplified.

[0065] The preferred outer membrane protein is MOMP but may also include other surface exposed antigens from Chlamydia species that are targets for humoral responses.

[0066] The immuno-repeat from a surface exposed region can be from the same serotype (homologous immuno-repeats) or represent fragments that contain variable epitopes and are derived from different serotypes (heterologous immuno-repeat). In a preferred embodiment the immuno-repeats contain an extended fragment that contains both a variable and a conserved region known to be rich in T cell epitopes.

[0067] A preferred surface exposed region of an outer membrane protein is chosen from VD1, VD2, VD3 and VD4 from MOMP.

[0068] The amino acid sequences used for constructing the immuno-repeats described in the examples are chosen from table 1, 2 and 3.

[0069] The variable domain of VD4 of MOMP can be described as an amino acid sequences as defined as:

La1-Aa2-Aa1-Aa3-La2

wherein Aa1 consists of the amino acid sequence TTLNPTIAG (SEQ ID NO 76) (which is conserved for all serovars); Aa2 is selected from the group consisting of: SATAIFDT (SEQ ID NO 79) (from serovar D and E), LVTPWDI (SEQ ID NO 80) (from serovar F), LAKPVVDI (SEQ ID NO 81) (from serovar G) and LAEAILDV (SEQ ID NO: 82) (from serovar Ia and J).

[0070] When Aa2 is the sequence from serovar D or E, then Aa3 is selected from the sequences set forth in AGDVKTGAEGQLG (SEQ ID NO 83) (from serovar D) and AGDVKASAEGQLG (SEQ ID NO 84) (serovar E).

[0071] When Aa2 is the sequence from serovar F, then Aa3 is the sequence CGSVAGANTEGQIS (SEQ ID NO 85) (from serovar F).

[0072] When Aa2 is the sequence from serovar G, then Aa3 is the sequence CGSVVAANSEGQIS (SEQ ID NO 86) (from serovar G).

[0073] When Aa2 is the sequence from serovar Ia or J), then Aa3 is selected from KGTVVSSAENELA (SEQ ID NO 87) (from serovar Ia) and KGTVVASGSENDLA (SEQ ID NO 88) (from serovar J)

[0074] The variable domain VD4 of MOMP is depicted in FIG. 2. The immuno-repeats preferably additionally comprises extensions on either sides which are also depicted in FIG. 2.

[0075] The N-terminal side of a VD4 domain can be flanked or extended by one or more amino acids from the more conserved and T-cell epitope rich La1, where La1 is the part of VD4 of MOMP which is embedded in the membrane and has the amino acid sequence EWQASLALSYRLNMFTPYIGVKWSRASFDADTIRIAQPK (SEQ ID NO 21) or an amino acid sequence having 80% sequence identity herewith.

[0076] The C-terminal side of a VD4 domain can correspondingly be flanked or extended by one or more amino acids from the more conserved and T-cell epitope rich La2, where La2 is the part of VD4 of MOMP which is embedded in the membrane on the C-terminal side and has the amino acid sequence DTMQIVSLQLNKMKSRKSCGIAVGTTIVDA (SEQ ID NO 22) or an amino acid sequence having 80% sequence identity herewith.

[0077] A similar illustration (see FIG. 1) can describe immuno-repeats comprising the variable domain 1 (VD1) of MOMP with the variable domains (Aa2-Aa1-Aa3) of the various serovars are given by SEQ ID NO 1-6 in table 1. The corresponding N-terminal and C-terminal extensions (La1 and La2) have the respective amino acid sequences SMRVGYYGDFVFDRVLKTDVNKEFQMG (SEQ ID NO 77) (La1) and NPAYGRHMQDAEMFTNAACMALNIWD (SEQ ID NO 78) (La2) which are given in table 2 by SEQ ID NO 7-8.

[0078] Immuno-repeats comprising VD2 and VD3 can in a similar manner be deduced from FIG. 1 and table 1.

[0079] Hence above example La1-Aa2-Aa1-Aa3-La2 defines one of the immune-repeat units. If additionally e.g. VD1 is added to a VD4 unit, this can be described as adding one more sequence to make up a larger immune-repeat unit. Hence the polypeptide of the invention comprises 2, 3, 4, 5, 6, 7 or 8 repeats of immune-repeat units.

DEFINITIONS

Outer Membrane Proteins

[0080] The outer membrane of Chlamydia sp. can be isolated by treating intact, purified elementary bodies with detergent such as 2% Sarkosyl followed by ultracentrigation (100,000 g for one hour) which will lead to a supernatant with cytosolic components and a pellet containing the outer membrane as previously described⁷⁰. Outer membrane proteins can then be identified by standard protein techniques, e.g. by mass spectrometry after SDS-PAGE.

Surface Exposed Fragments or Regions

[0081] Bacterial surface or membrane proteins comprises trans membrane proteins, secretory and lipoproteins, and anchorless surface proteins. Surface exposed regions on intact bacteria are accessible to antibodies. Methods to identify surface exposed regions of proteins (the `surfaceome` comprise e.g. biotinylation of the membrane proteins in intact bacteria, followed by isolation of the biotin-labelled fraction using streptavidin. The isolated proteins can then be identified by mass spectrometry. Another approach is to treat intact bacteria with a protease, e.g. trypsin (`shaving`) to cleave surface exposed peptides, followed by collection of the released peptides for identification by mass spectrometry.

Variants

[0082] Variants of outer membrane proteins provided herein describes proteins encoded by the same gene from different serotypes of Chlamydia sp. A variant protein shares significant homology with a reference polypeptide.

An Isoform of Protein

[0083] In the context of the present application an "isoform" of protein is under stood as any of several different forms of the same protein e.g. a protein that has the same function but which is encoded by a different gene and may have small differences in its sequence or arises from either single nucleotide polymorphisms, differential splicing of mRNA, or post-translational modifications. Different serotypes of bacteria may have different isoforms of certain proteins.

Chlamydia Species

[0084] By the term "Chlamydia species" is understood a bacterium capable of causing the Chlamydia infection in an animal or in a human being. Examples are C. trachomatis, C. pneumoniae and C. muridarum. Also in animals, several infections with Chlamydia sp. are known, e.g. Chlamydia Suis infecting pigs, and Chlamydiaphila abortus which causes abortion in small ruminants (sheep and goats).

Serovariants, Serovars or Serotypes

[0085] Based on the reactivity of specific mono clonal antibodies against and detailed sequence analysis of the MOMP variable regions Ct can be divided into 15 different serovariants and of these serovariants A, B, Ba and C causes Trachoma, D-K causes sexually transmitted disease (STD), L1-L3 causes Lymphogranuloma venerum, and MoPn (C. muridarum) infects mice. Serovariants are sometimes mentioned as serovars or serotypes with the same meaning.

Immuno-Repeats

[0086] By immuno-repeats is understood: repetitive units of one or more amino acid sequences comprising an immunogenic portion or fragment of an antigen. The units that are repeated can be described as one or more VD regions, that optionally can be extended as described above, that are repeated e.g. 4 examples with three repeats VD4-VD4-VD4, VD4-VD1-VD4-VD1-VD4-VD1, VD4_D-VD4_D-VD4_D, VD4_D-VD4_F-VD4.sub.G, VD4_D-VD3_E-VD4_D-VD3_E-VD4_D-VD3_E.

Homologous Immuno-Repeat

[0087] Repetitive units of one or more amino acid sequences comprising an immunogenic portion or fragment of an antigen from one serovariant only (FIG. 4)

Heterologous Immuno-Repeat

[0088] Repetitive units of one or more amino acid sequences comprising an immunogenic portion or fragment encoding the same antigen derived from different serovariants (FIG. 4).

Heterologous Challenge

[0089] Refers to the situation where the protein used for vaccination is derived from a different bacterial serovariant than the serovariant used for challenge.

Homologous Challenge

[0090] Refers to the situation where the protein used for vaccination is derived from the same bacterial serovariant as the serovariant used for challenge.

MOMP

[0091] The Major Outer Membrane Protein (MOMP) of Ct, is expressed during all phases of the developmental life cycle of Ct and constitutes approximately 60% of the total protein content of the chlamydia outer membrane. MOMP can be divided into conserved domains interrupted by four highly variable domains (VD1-4 or VS1-4)⁵⁹ (FIG. 1)

VD1

[0092] Variable domain 1 (VD1) of MOMP as defined by Baehr et al (1988)³⁶ which corresponds to amino acids 91-105 and make up a highly variable region in MOMP from Ct (Seq no 1-6 VD1 from SvD, E, F, G, Ia and J respectively). The extended VD1 region (VD1^ext) corresponds to amino acids 57-115 and make-up said highly variable region flanked by highly conserved regions in MOMP from Ct (Seq no 9-14 VD1^ext from SvD, E, F, G, Ia and J respectively) (FIG. 3).

VD4

[0093] Variable domain 4 of MOMP as defined by Baehr et al (1988)³⁶ which corresponds to amino acids 309-338 and make up a highly variable region in MOMP from Ct (Seq no 15-20 VD4 from SvD, E, F, G, Ia and J respectively). The extended VD4 region (VD4^ext) corresponds to amino acids 282-349 and make-up said highly variable region flanked by highly conserved regions in MOMP from Ct (Seq no 23-28 VD4^ext from SvD, E F, G, Ia and J respectively).

Linearized

[0094] The word "linearized" in the present invention refers to an amino acid chain of any length, including a full-length protein, oligopeptides, short peptides and fragments thereof, wherein the amino acid cysteine has been substituted with serine in order to hinder the cysteine residues to form disulfide bonds.

Neutralizing Epitope

[0095] Neutralizing epitope as used herein is intended an amino acid sequence that defines an antigenic determinant which is bound by an antibody and, in the context of infection, reduces infectivity of a Chlamydial load, e.g. by blocking of the bacterial interaction with host cells, which is important in establishing bacterial infection and disease, facilitating bacterial clearance.

Neutralization

[0096] Neutralization is to encompass any biological activity of the bacteria, including reduction in the efficiency or ability of the bacterium to establish infection or cause disease or disease symptoms, inhibition of chlamydial EB formation.

Neutralizing Antibodies

[0097] Antibodies which bind a neutralizing epitope as described above.

Polypeptides

[0098] The word "polypeptide" in the present invention should have its usual meaning. That is an amino acid chain of any length, including a full-length protein, oligopeptides, short peptides and fragments thereof, wherein the amino acid residues are linked by covalent peptide bonds.

IFN-γ

[0099] By the term "IFN-γ" is understood interferon-gamma. The measurement of IFN-γ is used as an indication of an immunological T-cell response.

Comprise

[0100] Throughout this specification, unless the context requires otherwise, the word "comprise", or variations thereof such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element or integer or group of elements or integers but not the exclusion of any other element or integer or group of elements or integers.

Immunogenic Portion or Fragment

[0101] In a preferred embodiment of the invention, the polypeptide comprises an immunogenic portion or fragment of the polypeptide, such as an epitope for a B-cell or T-cell. The immunogenic portion or fragment of a polypeptide is a part of the polypeptide, which elicits an immune response in an animal or a human being, and/or in a biological sample determined by any of the biological assays described herein. The immunogenic portion or fragment of a polypeptide may be a T-cell epitope or a B-cell epitope. Immunogenic portions or fragments can be related to one or a few relatively small parts of the polypeptide, they can be scattered throughout the polypeptide sequence or be situated in specific parts of the polypeptide. For a few polypeptides epitopes have even been demonstrated to be scattered throughout the polypeptide covering the full sequence.

[0102] In order to identify relevant T-cell epitopes which are recognised during an immune response, it is possible to use a "brute force" method: Since T-cell epitopes are linear, deletion mutants of the polypeptide will, if constructed systematically, reveal what regions of the polypeptide are essential in immune recognition, e.g. by subjecting these deletion mutants e.g. to the IFN-γ assay described herein. Another method utilises overlapping oligopeptides for the detection of MHC class II epitopes, preferably synthetic, having a length of e.g. 20 amino acid residues derived from the polypeptide. These peptides can be tested in biological assays (e.g. the IFN-γ assay as described herein) and some of these will give a positive response (and thereby be immunogenic) as evidence for the presence of a T cell epitope in the peptide. For the detection of MHC class I epitopes it is possible to predict peptides that will bind⁷² and hereafter produce these peptides synthetic and test them in relevant biological assays e.g. the IFN-γ assay as described herein. The peptides preferably having a length of e.g. 8 to 11 amino acid residues derived from the polypeptide. B-cell epitopes can be determined by analysing the B cell recognition to overlapping peptides covering the polypeptide of interest as e.g. described in Harboe et al⁷³.

Immunogenic

[0103] An immunogenic polypeptide is defined as a polypeptide that induces an immune response in a biological sample or an individual currently or previously infected with a chlamydia.

Fusion Proteins

[0104] By a fusion protein is understood two or more polypeptides linked together covalently. The fusion proteins can be produced with superior characteristics of the polypeptide. For instance, fusion partners that facilitate export of the fusion protein when produced recombinantly (e.g. signal peptides), fusion partners that facilitate purification of the fusion protein (e.g. his-tags), and fusion partners which enhance the immunogenicity of the fusion protein are all interesting possibilities. The fusion partner can, in order to enhance immunogenicity, be another polypeptide derived from C. trachomatis, such as a polypeptide, a polypeptide fragment or at least one T-cell epitope or B cell epitope.

Pharmaceutical Composition

[0105] A pharmaceutical composition is defined as any vaccine (both therapeutic and prophylactic) or any diagnostic reagent.

Vaccine, Protein

[0106] Another part of the invention pertains to a vaccine composition comprising a fusion protein or a nucleic acid encoding said fusion protein according to the invention. In order to ensure optimum performance of such a vaccine composition it is preferred that it comprises an immunologically and pharmaceutically acceptable carrier, vehicle or adjuvant.

[0107] An effective vaccine, wherein a fusion protein of the invention is recognized by a mammal including a human being, will decrease bacterial load in target organs, prolong survival times and/or diminish weight loss after challenge with virulent chlamydial bacteria, compared to non-vaccinated individuals.

[0108] Suitable carriers are selected from the group consisting of a polymer to which the polypeptide(s) is/are bound by hydrophobic non-covalent interaction, such as a plastic, e.g. polystyrene, or a polymer to which the polypeptide(s) is/are covalently bound, such as a polysaccharide, or a polypeptide, e.g. bovine serum albumin, ovalbumin or keyhole limpet haemocyanin. Suitable vehicles are selected from the group consisting of a diluent and a suspending agent. The adjuvant is preferably selected from the group consisting of dimethyl-dioctadecylammonium bromide (DDA), Quil A, poly I:C, aluminium hydroxide, Freund's incomplete adjuvant, IFNγ, IL-2, IL-12, monophosphoryl lipid A (MPL), Trehalose Dimycolate (TDM), Trehalose Dibephenate (TDB) and muramyl dipeptide (MDP), Monomycolyl glycerol (MMG) or a combination hereof. A preferred combination is a cationic liposome such as DDA combined with TDB and/or poly I:C.

[0109] Preparation of vaccines which contain peptide sequences as active ingredients is generally well understood in the art, as exemplified by U.S. Pat. Nos. 4,608,251; 4,601,903; 4,599,231 and 4,599,230, all incorporated herein by reference.

Therapeutic Vaccine.

[0110] The invention also relates to the use of a polypeptide or nucleic acid of the invention for use as therapeutic vaccines as have been described in the literature exemplified by D. Lowry (Lowry et al 1999). Antigens with therapeutic properties may be identified based on their ability to diminish the severity of Ct infection in experimental animals or prevent reactivation of previous infection, when administered as a vaccine. The composition used for therapeutic vaccines can be prepared as described above for vaccines.

[0111] The present invention describes novel highly immunogenic vaccine antigens with broad antibody based neutralizing capacity that protects against different serovariants of Chlamydia trachomatis. We demonstrate that repetitive units of defined fragments from the MOMP antigen provide highly immunogenic molecules which we refer to as immuno-repeats. Vaccination with homologous immuno-repeats containing VD4 extended fragments (covers the VD4 variable domain of MOMP and the adjacent conserved flanking regions) in different adjuvants provides very high antibody titers and we demonstrate that these constructs are much more efficient than immunizing with single units of the VD4 extended fragment. The increased effect can be observed both as markedly increased titer, increased antibody targeting of the surface of the bacteria, increased neutralizing capacity, increased and broadened T cell response and increased protection against a challenge with the homologous strain. We furthermore demonstrate that the immuno-repeat technology can be utilized also to improve the protection against and neutralization of other serovariants by constructing heterologous immuno-repeats based on VD4 extended fragments from different serovariants such as serovar D, E, F and G (FIG. 3).

TABLE-US-00001 TABLE 1 Description of sequences used in constructing immuno-repeats SEQ ID NO Variable domaines Description 1 VD1_SvD Serovar D variable domaine 1 of MOMP 2 VD1_SvE Serovar E variable domaine 1 of MOMP 3 VD1_SvF Serovar F variable domaine 1 of MOMP 4 VD1_SvG Serovar G variable domaine 1 of MOMP 5 VD1_SvIa Serovar Ia variable domaine 1 of MOMP 6 VD1_SvJ Serovar J variable domaine 1 of MOMP 7 VD1 N-terminal VD1 N-terminal extension 8 VD1 C-terminal VD1 C-terminal extension 9 VD1ext_SvD Serovar D extended VD1 of MOMP 10 VD1ext_SvE Serovar E extended VD1 of MOMP 11 VD1ext_SvF Serovar F extended VD1 of MOMP 12 VD1ext_SvG Serovar G extended VD1 of MOMP 13 VD1ext_SvIa Serovar Ia extended VD1 of MOMP 14 VD1ext_SvJ Serovar J extended VD1 of MOMP 15 VD4_SvD Serovar D variable domaine 4 of MOMP 16 VD4_SvE Serovar E variable domaine 4 of MOMP 17 VD4_SvF Serovar F variable domaine 4 of MOMP 18 VD4_SvG Serovar G variable domaine 4 of MOMP 19 VD4_SvIa Serovar Ia variable domaine 4 of MOMP 20 VD4_SvJ Serovar J variable domaine 4 of MOMP 21 VD4 N-terminal VD4 N-terminal extension 22 VD4 C-terminal VD4 C-terminal extension 23 VD4ext_SvD Serovar D extended VD4 of MOMP 24 VD4ext_SvE Serovar E extended VD4 of MOMP 25 VD4ext_SvF Serovar F extended VD4 of MOMP 26 VD4ext_SvG Serovar G extended VD4 of MOMP 27 VD4ext_SvIa Serovar Ia extended VD4 of MOMP 28 VD4ext_SvJ Serovar J extended VD4 of MOMP 29 VD2_SvD Serovar D variable domaine 2 of MOMP 30 VD2_SvE Serovar E variable domaine 2 of MOMP 31 VD2_SvF Serovar F variable domaine 2 of MOMP 32 VD2_SvG Serovar G variable domaine 2 of MOMP 33 VD2_SvIa Serovar Ia variable domaine 2 of MOMP 34 VD2_SvJ Serovar J variable domaine 2 of MOMP 35 VD2 N-terminal VD2 N-terminal extension 36 VD2 C-terminal VD2 C-terminal extension 37 VD3_SvD Serovar D variable domaine 3 of MOMP 38 VD3_SvE Serovar E variable domaine 3 of MOMP 39 VD3_SvF Serovar F variable domaine 3 of MOMP 40 VD3_SvG Serovar G variable domaine 3 of MOMP 41 VD3_SvIa Serovar Ia variable domaine 3 of MOMP 42 VD3_SvJ Serovar J variable domaine 3 of MOMP 43 VD3 N-terminal VD3 N-terminal extension 44 VD3 C-terminal VD3 C-terminal extension

[0112] Heterologous immuno-repeats were highly immunogenic but in addition increased the breadth of the antibody responses which was associated with a broader fine specificity of the antibody response (measured by peptide scans) that targets a more diverse repertoire of linear epitopes within the VD4 region than the homologous immuno-repeats. We also demonstrate that highly immunogenic heterologous immuno-repeats can be based on even larger fragments that incorporate fusions of VD1 and VD4 extended fragments and we confirm that in animal models protection promoted by these heterologous immuno-repeats are mediated predominantly by antibodies. As there is a generally recognized need for a strong CMI component (e.g. a T-cell epitope) in an efficient protective immune response against Ct, we have also demonstrated that by fully extending the VD4 region N-terminally to include a T cell rich region, we can generate immune-repeats that combine the ability to generate high tittered neutralizing antibodies with a strong T cell response clearing residual infection in one construct. We have also demonstrated that immune-repeats can be fused to or mixed with T-cell antigens with vaccine potential and that this combination provide both an early antibody mediated protection against Ct as well as an efficient CMI mediated clearance of residual organisms.

[0113] MOMP is an important protective antigen with a generally recognized potential in Ct vaccines. The MOMP antigen is however a very complicated antigen to target by vaccines because it has a complex structure with numerous internal disulfide bonds and where important neutralizing epitopes have been exceedingly difficult to expose in recombinant molecules. Adding to this, the MOMP antigen is highly variable and is the basis for the majority of the serovariance found in different strains causing human disease. Any vaccine based on intact MOMP would therefore have to incorporate a number of different versions of the molecule (at least 4-5) to cover the major strains giving rise to disease in humans. As described above the MOMP antigen contains 4 variable regions (VD1-4) of which in particular the VD1 and VD4 contain important neutralizing epitopes but vaccines based on fragments representing these regions have so far failed to induce sufficiently high titers of functional antibodies to have any in vivo effect in animal challenge studies⁵¹ 74.

[0114] The immuno-repeat technology of the present invention solves this problem: By repeating the important variable VD1 and/or VD4 regions flanked by conserved sequences from the MOMP antigen we have obtained immunogens that promote extraordinary levels of functional antibodies. Surprisingly we also demonstrate that the improved immunogenicity can even be achieved in heterologous immuno-repeat constructs that employs variable regions from different serovars interspaced between conserved fragments and that this strategy produces a broadly neutralizing antibody response that protect against different serovariants. Furthermore, do the immuno-repeat technology provide a large number of relevant T cell epitopes that promote T cells with direct effector function as well as the ability to promote accelerated recall responses to the adjacent B cell epitopes.

[0115] Our invention therefore represents a breakthrough in developing efficient Ct vaccines with a broad response and the ability to neutralize different serovars.

[0116] It is well known that antigens with a large number of repeats and organized structure are optimal for the activation of the B-cell receptor (BCR), leading to an increased humoral response and a decreased dependence on T-cell help. This was originally reported with natural polysaccharide based antigens from various pathogens (Pneumococcal polysaccharide and Salmonella polymerized flagellin) where the repetitive nature of the antigen is assumed to trigger several BCR simultaneously thereby lowering the overall activation threshold which triggers antibody production from plasma B-cells without the need for prior T-cell help. Such antigens are referred to as type 2 T-cell independent B-cell antigens and in artificial systems have been shown to depend on a large number of repeats (typically a minimum of 12-16⁷⁵), that constitute the minimal epitope and are closely located. This is clearly different from our repeat technology where large fragments (69 amino acids, Mw>7 kDa) are repeated and these fragments contain both B-cell and T-cell epitopes⁷⁶.

[0117] In contrast to previous observations⁷⁵, we observe an increase by just 4 repeats which is not further improved by 8 repeats. Importantly, the repetition of a conserved sequence with hypervariable domains inserted, amplify responses not only to the repeated conserved element but importantly to the variable inserts. The molecular mechanism behind this surprising amplification is not completely clear but it most likely relates to the fact that many of the important epitopes are located in the overlap between variable and conserved regions which therefore may allow simultaneous triggering of different BCR's that all share some recognition of the conserved part of the epitope. Although the mechanism is not completely clear the practical consequence is that the heterologous immune-repeat technology allows the synthesis of a multivalent immunogens that promote the generation of a diverse antibody response that targets different serovariants.

[0118] Our immuno-repeat constructs provide antigens of an extraordinary immunogenicity compared to previous attempts to use the variable domains from Ct MOMP. All previous vaccines based on VDs of MOMP did, in spite of generating antibodies with some functional capabilities, fail to generate titres that translated into in vivo protection against genital chlamydial challenge⁵¹, 65 64. In particular the heterologous immuno-repeat strategy solves a very fundamental problem seen for many pathogens and that is how to promote diverse antibody responses to diverse and variable antigens.

TABLE-US-00002 TABLE 2 Immuno-repeats SEQ ID NO Polypeptide names Description 45 CTH87 (CT681_VD1ext_VD4ext_SvD) Fusion of VD1-VD4 of serovar D 46 CTH88 (CT681_lin_VD1ext_VD4ext_SvD_E_F) Heterologous immune repeat of VD1-VD4 47 CTH88ext = CTH69 Same as SEQ ID NO 46 with longer (CT681_lin_VD1ext_VD4ext_SvD_E_F_ext) flanking region. 48 CTH72 Same as seq id no 47 additionally with (CT681_lin_VD1ext_VD4ext_SvD_E_F_G_Ia_J_ext) VD1ext and VD4ext from SvG, SvIa and SvJ 49 CTH89 (CT681_lin_VD4ext_SvD_E_F) Heterologous immune repeat of VD4 50 CTH181 (CT681_VD4ext_SvE) Same as SEQ ID NO 24 51 CTH182 (CT681_lin_VD4ext_F) Same as SEQ ID NO 25 linearized 52 CTH183 (CT681_VD4ext_F) Same as SEQ ID NO 25 53 CTH518 (CT681_Lin_VD4ext_D_E_F_G) Heterologous immune repeat of VD4 54 CTH518ext = CTH70 Same as SEQ ID NO 53 with longer (CT681_lin_VD4ext_SvD_E_F_G_ext) flanking regions 55 CTH71 Same as seq id no 54 additionally with (CT681_lin_VD4ext_SvD_E_F_G_Ia_J_ext) VD1ext and VD4ext from SvIa and SvJ 56 CTH524 (CT681_lin_4_VD4ext_F) Same as SEQ ID NO 59 linearized 57 CTH526 (CT681_8_VD4ext_SvE) Homologous immune repeat of VD4 (8x) 58 CTH527 (CT681_4_VD4ext_SvE) Homologous immune repeat of VD4 (4x) 59 CTH529 (CT681_4_VD4ext_F) Homologous immune repeat of VD4 (4x)

TABLE-US-00003 TABLE 3 Examples of immuno-repeats fused with T-cell antigens SEQ ID NO Fusions of immuno repeats with T-cell antigens (all his-tagged) 60 CTH91 (CT043-CT414p-CT681_lin_VD1ext_VD4ext_SvD_E_F) 61 CTH93 (CT043 CT414p_CT681_Lin_56-281_VD4ext_D) 62 CTH520 (CT681_56-281_VD4ext_D) 63 CTH521 (CT681_Lin_56-281_VD4ext_D) 64 CTH522 (CT681_lin_56-281_VD4ext_D_E_F_G) 65 CTH531 (CT414_CT043_CT043_681_lin_56-281_VD4ext_SvD_E_F_G) 66 CTH533 (CT043_CT043_CT681_lin_VD4ext_SvD_E_F_G) 67 CTH534 (CT043_CT043_CT004_CT681_lin_VD4ext_SvD_E_F_G) 68 CT681_SvD 69 CTH285 (VD4_lin_SvD, E, F, G) 70 CTH286 (VD4_classic + 7_lin_SvD, E, F, G)

TABLE-US-00004 TABLE 4 Overlapping peptides of VD4 from serovarE VD4 serovar E peptides (20mers) Amino acid sequence SEQ ID NO CT681_25_SvE DASIDYHEWQASLALSYRLN 89 CT681_26_SvE ASLALSYRLNMFTPYIGVKW 90 CT681_27_SvE MFTPYIGVKWSRASFDADTI 91 CT681_28_SvE SRASFDADTIRIAQPKSATA 92 CT681_29_SvE RIAQPKSATAIFDTTTLNPT 93 CT681_30_SvE IFDTTTLNPTIAGAGDVKAS 94 CT681_31_SvE IAGAGDVKASAEGQLGDTMQ 95 CT681_32_SvE AEGQLGDTMQIVSLQLNKMK 96

TABLE-US-00005 TABLE 5 Overlapping peptides of VD4 from serovarF Serovar F peptides (20mers) Amino acid sequence SEQ ID NO CT681_25_SvF DASIDYHEWQASLSLSYRLN 97 CT681_26_SvF ASLSLSYRLNMFTPYIGVKW 98 CT681_27_SvF MFTPYIGVKWSRASFDSDTI 99 CT681_28_SvF SRASFDSDTIRIAQPRLVTP 100 CT681_29_SvF RIAQPRLVTPVVDITTLNPT 101 CT681_30_SvF VVDITTLNPTIAGCGSVAGA 102 CT681_31_SvF IAGCGSVAGANTEGQISDTMQ 103 CT681_32_SvF TEGQISDTMQIVSLQLNKMK 104

TABLE-US-00006 TABLE 6 Overlapping peptides of VD4 from serovar D VD4 serovar D Amino acid VD4 serovar D Amino acid peptides (9mers) sequence SEQ ID NO peptides (9mers) sequence SEQ ID NO VD4_P1_SvD SRASFDADT 105 VD4_P24_SvD TTTLNPTIA 128 VD4_P2_SvD RASFDADTI 106 VD4_P25_SvD TTLNPTIAG 76 VD4_P3_SvD ASFDADTIR 107 VD4_P26_SvD TLNPTIAGA 129 VD4_P4_SvD SFDADTIRI 108 VD4_P27_SvD LNPTIAGAG 130 VD4_P5_SvD FDADTIRIA 109 VD4_P28_SvD NPTIAGAGD 131 VD4_P6_SvD DADTIRIAQ 110 VD4_P29_SvD PTIAGAGDV 132 VD4_P7_SvD ADTIRIAQP 111 VD4_P30_SvD TIAGAGDVK 133 VD4_P8_SvD DTIRIAQPK 112 VD4_P31_SvD IAGAGDVKT 134 VD4_P9_SvD TIRIAQPKS 113 VD4_P32_SvD AGAGDVKTG 135 VD4_P10_SvD IRIAQPKSA 114 VD4_P33_SvD GAGDVKTGA 136 VD4_P11_SvD RIAQPKSAT 115 VD4_P34_SvD AGDVKTGAE 137 VD4_P12_SvD IAQPKSATA 116 VD4_P35_SvD GDVKTGAEG 138 VD4_P13_SvD AQPKSATAI 117 VD4_P36_SvD DVKTGAEGQ 139 VD4_P14_SvD QPKSATAIF 118 VD4_P37_SvD VKTGAEGQL 140 VD4_P15_SvD PKSATAIFD 119 VD4_P38_SvD KTGAEGQLG 141 VD4_P16_SvD KSATAIFDT 120 VD4_P39_SvD TGAEGQLGD 142 VD4_P17_SvD SATAIFDTT 121 VD4_P40_SvD GAEGQLGDT 143 VD4_P18_SvD ATAIFDTTT 122 VD4_P41_SvD AEGQLGDTM 144 VD4_P19_SvD TAIFDTTTL 123 VD4_P42_SvD EGQLGDTMQ 145 VD4_P20_SvD AIFDTTTLN 124 VD4_P43_SvD GQLGDTMQI 146 VD4_P21_SvD IFDTTTLNP 125 VD4_P44_SvD QLGDTMQIV 147 VD4_P22_SvD FDTTTLNPT 126 VD4_P45_SvD LGDTMQIVS 148 VD4_P23_SvD DTTTLNPTI 127

TABLE-US-00007 TABLE 7 Overlapping peptides of VD4 from serovar F VD4 serovar F Amino acid VD4 serovar F Amino acid peptides (9mers) sequence SEQ ID NO peptides (9mers) sequence SEQ ID NO VD4_P1_SvF SRASFDSDT 149 VD4_P24_SvF ITTLNPTIA 172 VD4_P2_SvF RASFDSDTI 150 VD4_P25_SvF TTLNPTIAG 76 VD4_P3_SvF ASFDSDTIR 151 VD4_P26_SvF TLNPTIAGC 173 VD4_P4_SvF SFDSDTIRI 152 VD4_P27_SvF LNPTIAGCG 174 VD4_P5_SvF FDSDTIRIA 153 VD4_P28_SvF NPTIAGCGS 175 VD4_P6_SvF DSDTIRIAQ 154 VD4_P29_SvF PTIAGCGSV 176 VD4_P7_SvF SDTIRIAQP 155 VD4_P30_SvF TIAGCGSVA 177 VD4_P8_SvF DTIRIAQPR 156 VD4_P31_SvF IAGCGSVAG 178 VD4_P9_SvF TIRIAQPRL 157 VD4_P32_SvF AGCGSVAGA 179 VD4_P11_SvF IRIAQPRLV 158 VD4_P33_SvF GCGSVAGAN 180 VD4_P11_SvF RIAQPRLVT 159 VD4_P34_SvF CGSVAGANT 181 VD4_P12_SvF IAQPRLVTP 160 VD4_P35_SvF GSVAGANTE 182 VD4_P13_SvF AQPRLVTPV 161 VD4_P36_SvF SVAGANTEG 183 VD4_P14_SvF QPRLVTPVV 162 VD4_P37_SvF VAGANTEGQ 184 VD4_P15_SvF PRLVTPVVD 163 VD4_P38_SvF AGANTEGQI 185 VD4_P16_SvF RLVTPVVDI 164 VD4_P39_SvF GANTEGQIS 186 VD4_P17_SvF LVTPVVDIT 165 VD4_P40_SvF ANTEGQISD 187 VD4_P18_SvF VTPVVDITT 166 VD4_P41_SvF NTEGQISDT 188 VD4_P19_SvF TPVVDITTL 167 VD4_P42_SvF TEGQISDTM 189 VD4_P20_SvF PVVDITTLN 168 VD4_P43_SvF EGQISDTMQ 190 VD4_P21_SvF VVDITTLNP 169 VD4_P44_SvF GQISDTMQI 191 VD4_P22_SvF VDITTLNPT 170 VD4_P45_SvF QISDTMQIV 192 VD4_P23_SvF DITTLNPTI 171 VD4_P46_SvF ISDTMQIVS 193

TABLE-US-00008 TABLE 8 CT681 amino acid sequences SEQ Amino acid sequences of MOMP ID NO (CT681) from different serovars 68 CT681_SvD 71 CT681_SvE 72 CT681_SvF 73 CT681_SvG 74 CT681_SvIa 75 CT681_SvJ

[0119] The nucleic acid of the invention, that is nucleic acid encoding above mentioned fusion proteins, may be used for effecting in vivo expression of immunogenic polypeptides, i.e. the nucleic acid may be used in so-called DNA vaccines as reviewed in Ulmer et al 1993, which is included by reference.

[0120] In the construction and preparation of plasmid DNA encoding a fusion polypeptide to be used defined for DNA vaccination a host strain such as E. coli can be used. Plasmid DNA can then be prepared from overnight cultures of the host strain carrying the plasmid of interest, and purified using e.g. the Qiagen Giga-Plasmid column kit (Qiagen, Santa Clarita, Calif., USA) including an endotoxin removal step. It is essential that plasmid DNA used for DNA vaccination is endotoxin free.

[0121] Hence, the invention also relates to a vaccine comprising a nucleic acid according to the invention, the vaccine effecting in vivo expression of the immunogenic polypeptide by an animal, including a human being, to whom the vaccine has been administered, the amount of expressed polypeptide being effective to confer substantially increased resistance to infections caused by virulent bacteria in an animal, including a human being.

[0122] The efficacy of such a DNA vaccine can possibly be enhanced by administering the gene encoding the expression product together with a DNA fragment encoding a polypeptide which has the capability of modulating an immune response.

[0123] One possibility for effectively activating a cellular immune response can be achieved by expressing the relevant immunogenic polypeptide in a non-pathogenic microorganism or virus. Well-known examples of such microorganisms are Mycobacterium bovis BCG, Salmonella and Pseudomona and examples of viruses are Vaccinia Virus and Adenovirus.

[0124] Therefore, another important aspect of the present invention is an improvement of the live BCG vaccine presently available, wherein one or more copies of a DNA sequence encoding one or more fusion polypeptides as defined above has been incorporated into the genome of the micro-organism in a manner allowing the micro-organism to express and secrete the fusion polypeptide. The incorporation of more than one copy of a nucleic acid sequence of the invention is contemplated to enhance the immune response.

[0125] Another possibility is to integrate the DNA encoding the fusion polypeptide according to the invention in an attenuated virus such as the Vaccinia virus or Adenovirus (Rolph et al 1997). The recombinant vaccinia virus is able to enter within the cytoplasma or nucleus of the infected host cell and the fusion polypeptide of interest can therefore induce an immune response, which is envisioned to induce protection against T B.

[0126] Although DNA vaccines were developed more than 16 years ago, clinical trials preceding stage I and II in humans are rare. Two veterinary DNA vaccines however, have been licensed; one for West Nile Virus (in horse) and a second for Infectious Hematopoetic Necrosis virus in Salmon. This demonstrates that DNA vaccines can have good protective effects and that new DNA vaccines are not limited by the size of the animal or species. The great success with DNA vaccines observed for the murine model for first generation DNA vaccines did not translate well to humans, nonetheless; researchers have recently demonstrated protective antibodies levels by a single dose of gene gun administrated HA DNA vaccine to humans.

[0127] "Nucleic acid immunization" or the commonly preferred name "DNA vaccines" are the inoculation of antigen encoding DNA or RNA as expression cassettes or expression vectors or incorporated into viral vectors with the purpose of inducing immunity to the gene product. Thus, in our definition of DNA vaccines we include all kinds of delivery systems for the antigen encoding DNA or RNA. The vaccine gene can be in form of circular plasmid or a linear expression cassette with just the key features necessary for expression (promotor, the vaccine gene and polyadenylation signal). Delivery systems may most often be naked DNA in buffer with or without adjuvant, DNA coupled to nanoparticles and/or formulated into adjuvant containing compounds or inserted into live viral or bacterial vectors such as Adenovirus, adeno associated virus, alphavirus, poxviruses, herpes virus etc. DNA vaccines hold great promise since they evoke both humoral and cell-mediated immunity, without the same dangers associated with live virus vaccines. In contrast to live attenuated virus vaccines DNA vaccines may be delivered to same or different tissue or cells than the live virus that has to bind to specific receptors. The production of antigens in their native forms improves the presentation of the antigens to the host immune system. Unlike live attenuated vaccines, DNA vaccines are not infectious and cannot revert to virulence.

[0128] DNA vaccines offer many advantages over conventional vaccines. It can be produced in high amounts in short time, abolishing the need for propagation in eggs, it is cost-effective, reproducible and the final product does not require cold storage conditions, because DNA is stable and resistant to the extremes of temperature. All currently licensed inactivated vaccines are efficient at inducing humoral antibody responses but only live attenuated virus vaccines efficiently induce a cytotoxic cellular response as well. DNA vaccines also have this ability and the induced response therefore may better mimic the natural response to viral infection than inactivated vaccines in respect to specificity and antibodies isotypes.

[0129] DNA vaccines induce an immune response which is comparable to the response acquired by natural virus infection by activating both humoral and cell-mediated immunity. The broad response to DNA vaccines is a result of the encoded genes being expressed by the transfected host cell, inducing both a Th1 and Th2 immune responses. The production of antigens in their native form improves the presentation of the antigens to the host immune system.

[0130] The two most common types of DNA vaccine administration are saline injection of naked DNA and gene gun DNA inoculations (DNA coated on solid gold beads administrated with helium pressure). Saline intra muscular injections of DNA preferentially generates a Th1 IgG2a response while gene gun delivery tends to initiate a more Th2 IgG1 response. Intramuscular injected plasmids are at risk of being degraded by extracellular deoxyribonucleases, however, the responses induced are often more long-lived than those induced by the gene gun method. Vaccination by gene gun delivery of DNA, to the epidermis, has proven to be the most effective method of immunization, probably because the skin contains all the necessary cells types, including professional antigen presenting cells (APC), for eliciting both humoral and cytotoxic cellular immune responses (Langerhans and dendritic cells). Complete protection from a lethal dose of influenza virus has been obtained with as little as 1 μg DNA in mice. The standard DNA vaccine vector consists of the gene of interest cloned into a bacterial plasmid engineered for optimal expression in eukaryotic cells. Essential features include; an origin of replication allowing for production in bacteria, a bacterial antibiotic resistance gene allowing for plasmid selection in bacterial culture, a strong constitutive promotor for optimal expression in mammalian cells (promoters derived from cytomegalovirus (CMV) or simian virus provide the highest gene expression), a polyadenylation sequence to stabilise the mRNA transcripts, such as bovine growth hormone (BHG) or simian virus polyadenylation, and a multiple cloning site for insertion of an antigen gene. An intron A sequence improves expression of genes remarkably. Many bacterial DNA vaccine vectors contain unmethylated cytidinephosphate-guanosine (CpG) dinucleotide motifs that may elicit strong innate immune responses in the host. In recent years there have been several approaches to enhance and customise the immune response to DNA vaccine constructs (2nd generation DNA vaccines). For instance dicistronic vectors or multiple geneexpressing plasmids have been used to express two genes simultaneously. Specific promoters have been engineered that restrict gene expression to certain tissues, and cytokine/antigen fusion genes have been constructed to enhance the immune response. Furthermore, genes may be codon optimised for optimal gene expression in the host and naive leader sequences may be substituted with optimised leaders increasing translation efficiency.

[0131] The administration of DNA vaccine can be by saline or buffered saline injection of naked DNA or RNA, or injection of DNA plasmid or linear gene expressing DNA fragments coupled to particles, or inoculated by gene gun or delivered by a viral vector (virus like particle) such as Adenovirus, Modified vaccinia virus Ankara (MVA), Vaccinia, Adenoassociated virus (AAV), Alphavirus etc.

[0132] In one embodiment is a polypeptide comprising

a) an amino acid sequence comprising one or more surface exposed fragments of the same outer membrane protein expressed in a serotype of Chlamydia sp.; and b) two or more additional amino acid sequences which is either the same sequence as defined in a) or is the corresponding surface exposed fragments from a variant of said outer membrane protein expressed in a serotype of Chlamydia sp., which is different from the serotype in a).

[0133] In a further embodiment is a polypeptide comprising 3 or more different amino acid sequences, where said amino acid sequences each comprises one or more surface exposed fragments from different variants of the same outer membrane protein that varies in different Chlamydia sp. serotypes, said amino acid sequences derived from different Chlamydia sp. serotypes.

[0134] In another further embodiment is a polypeptide comprising 3 or more repetitions of an amino acid sequence, where said amino acid sequence comprises one or more surface exposed fragments of the same outer membrane protein that varies in different Chlamydia sp. serotypes, said amino acid sequences derived from the same Chlamydia sp. serotype.

[0135] A polypeptide as described above is provided, wherein the outer membrane protein is MOMP from any serotype. The outer membrane protein may be MOMP from serotype D, E, F, G, Ia or J of Chlamydia trachomatis or C. pneumoniae. Still further, a polypeptide may comprise one or more of the variable domains 1, 2, 3, 4 of MOMP. These variable domain sequences may optionally be linearized. These variable domain sequences may comprise the variable domains 4 (VD4) of MOMP, and may be placed next to each other or be spaced with a linker. In an embodiment thereof is a polypeptide comprising an amino acid sequence defined in formula I:

xx₁-VD4-xx₂ (Formula I)

[0136] wherein

[0137] VD4 is independently selected from SEQ ID NO. 15-20 or an amino acid sequence which has at least 80% sequence identity herewith

[0138] and

[0139] xx₁ consists of

[0140] i) The amino acid sequence EWQASLALSYRLNMFTPYIGVKWSRASFDADTIRIAQPK (SEQ ID NO 21) or

[0141] ii) A subsequence of the amino acid sequence in i) said subsequence comprising 1-38 amino acid residues, starting with the C-terminal K in the amino acid sequence in i)

[0142] and

[0143] xx₂ consists of

[0144] iii) The amino acid sequence DTMQIVSLQLNKMKSRKSCGIAVGTTIVDA (SEQ ID NO 22) or

[0145] iv) A subsequence of the amino acid sequence in iii) said subsequence comprising 1-29 amino acid residues, starting with the N-terminal D in the amino acid sequence in iii).

[0146] In these embodiments, the sequences may be chosen from SEQ ID NO. 23-28, 49-59.

[0147] Polypeptides according to any of the above embodiments are also provided additionally comprising a fragment comprising the variable domains 1 (VD1) of MOMP and wherein the amino acid sequences comprising VD1 of MOMP are placed next to each other or are spaced with a linker. In an embodiment thereof is a polypeptide comprising an amino acid sequence defined in formula II:

yy₁-VD1-yy₂ (Formula II)

[0148] wherein

[0149] VD1 is independently selected from SEQ ID NO. 1-6 or an amino acid sequence which has at least 80% sequence identity herewith

[0150] and

[0151] yy₁ consists of

[0152] v) The amino acid sequence DAISMRVGYYGDFVFDRVLKTDVNKEFQMG SEQ ID NO 7) or

[0153] vi) A subsequence of the amino acid sequence in v) said subsequence comprising 1-30 amino acid residues, starting with the C-terminal G in the amino acid sequence in v)

[0154] and

[0155] yy₂ consists of

[0156] vii) The amino acid sequence NPAYGRHMQDAEMFTNAA (SEQ ID NO 8) or

[0157] viii) A subsequence of the amino acid sequence in vii) said subsequence comprising 1-18 amino acid residues, starting with the N-terminal N in the amino acid sequence in vii).

[0158] In these embodiments, the sequences may be chosen from SEQ ID NO. 9-14, 45-48.

[0159] Polypeptides according to any of the above embodiments are also provided comprising a fragment comprising the variable domains 2 (VD2) of MOMP and wherein the amino acid sequences comprising VD2 of MOMP are placed next to each other or are spaced with a linker. In an embodiment thereof is a polypeptide comprising an amino acid sequence defined in formula III:

zz₁-VD2-zz₂ (Formula III)

[0160] wherein

[0161] VD2 is independently selected from SEQ ID NO. 29-34 or an amino acid sequence which has at least 80% sequence identity herewith,

[0162] and

[0163] zz₁ consists of

[0164] ix) The amino acid sequence TLGATSGYLKGNSASFNLVGLFG (SEQ ID NO 35) or

[0165] x) A subsequence of the amino acid sequence in ix) said subsequence comprising 1-23 amino acid residues, starting with the C-terminal G in the amino acid sequence in ix)

[0166] and

[0167] xx₂ consists of

[0168] xi) The amino acid sequence VVELYTDTTFAWSVGARAALWE (SEQ ID NO 36) or

[0169] xii) A subsequence of the amino acid sequence in xi) said subsequence comprising 1-22 amino acid residues, starting with the N-terminal V in the amino acid sequence in xi).

[0170] Polypeptides according to any of the above embodiments are also provided comprising a fragment comprising the variable domains 3 (VD3) of MOMP and wherein the amino acid sequences comprising VD3 of MOMP are placed next to each other or are spaced with a linker. In an embodiment thereof is a polypeptide comprising an amino acid sequence defined in formula IV:

qq₁-VD3-qq₂ (Formula IV)

[0171] wherein

[0172] VD3 is independently selected from SEQ ID NO. 37-42 or an amino acid sequence which has at least 80% sequence identity herewith,

[0173] and

[0174] qq₁ consists of

[0175] xiii) The amino acid sequence ATLGASFQYAQSKPKVEELNVLCNAAEFTINKPKGYVG (SEQ ID NO 43) or

[0176] xiv) A subsequence of the amino acid sequence in xiii) said subsequence comprising 1-22 amino acid residues, starting with the C-terminal G in the amino acid sequence in xiii)

[0177] and

[0178] qq₂ consists of

[0179] xv) The amino acid sequence TGTKDASIDYHEWQASLALSYRLNMFTPYIGVKWS (SEQ ID NO 44) or

[0180] xvi) A subsequence of the amino acid sequence in xv) said subsequence comprising 1-35 amino acid residues, starting with the N-terminal T in the amino acid sequence in xv).

[0181] Polypeptides according to any of the above embodiments are also provided comprising a moiety that facilitate export of the polypeptide when produced recombinantly (e.g. signal peptides), a moiety that facilitate purification of the fusion protein (e.g. his-tags) and/or a moiety which enhance the immunogenicity (e.g. a T cell antigen). In some embodiments, the enhancer of immunogenicity is an additional T-cell target which is chosen from a Ct antigen such as CT043, CT004, CT414, CT681 or part hereof. In these embodiments, said sequences may be chosen from SEQ ID NO 60-68.

[0182] Still further provided are polypeptides according to any of the above embodiments, said polypeptide having the ability to

[0183] a) neutralize C. trachomatis serovar D in vitro with a 50% neutralization titer of 10^-3 or less, when tested in an experimental set-up comprising the administering heterologous immuno-repeats

[0184] b) neutralize C. trachomatis serovar D in vivo in at least 50% of the mice at day 7 post infection when tested in a mouse model comprising administering heterologous immuno-repeats

[0185] c) broaden the immune response to multiple serovars of C. trachomatis in vitro when administering a heterologous of immuno-repeats

[0186] Still further provided are nucleic acids encoding a polypeptides according to any of the above embodiments.

[0187] Also provided are pharmaceutical compositions comprising a polypeptide according to any of the above embodiments or a nucleic acid according to any of the above embodiments. The pharmaceutical compositions may be vaccines. The pharmaceutical compositions may additionally comprise a pharmacologically acceptable carrier, excipient, adjuvant or immune modulator. The pharmaceutical compositions may include an adjuvant selected from DDA/TDB or alum. In further embodiments, pharmaceutical compositions may include a carrier that is a virus-like particle.

[0188] Still further provided are pharmaceutical compositions comprising a polypeptide according to any of the above embodiments or a nucleic acid according to any of the above embodiments for prophylactic or therapeutic use against Chlamydia sp. infections, including infections with Chlamydia trachomatis or C. pneumoniae.

[0189] Methods for preventing, treating and/or reducing the incidence of Chlamydia sp.

[0190] infections, including infections with Chlamydia trachomatis and C. pneumoniae, said method comprising administering a pharmaceutical composition described herein are also provided.

Material and Methods

[0191] Cultivation of C. trachomatis

[0192] Ct serovar D, E and F was propagated in Hela 229 cells (ATCC, Rockville, Md., USA). The cells were cultivated in RPMI 1640 (Gibco BRL, Grand Island, N. Y., USA) media containing 5% fetal calf serum (Gibco BRL; heat inactivated), 1% v/v Hepes, 1% v/v L-glutamine, 1% v/v pyrovate and 10 μg/ml gentamycine. Semiconfluent monolayers of Hela 229 cells in 6 well-plates were infected with 1.5 inclusion forming unit per cell of Ct serovar E or F in 0.3 ml SPG-buffer/well. The plates were centrifuged 1 hour in a Heraeus Multifuge 3S at 750 g and incubated on a plate rocker for 2 h at 35° C. After 2 h 2 ml cultivation media supplemented with 5% glucose and 1 μg/ml cycloheximid were added pr. well and the cells were further incubated for 72 h at 37° C. in an atmosphere of 5% CO₂ in humidified air.

Harvesting of Ct

[0193] Chlamydiae were harvested 72 h post infection. The cells were dislodged from the wells with a cell scraper and centrifuged 30 minutes at 35.000 g and 4° C. The pellets were resuspended in HBSS, sonicated on ice and centrifuged at 500 g and 4° C. for 15 minutes. The supernatant was collected and saved on ice and the pellet was resuspended to same volume as before and sonication and centrifugation were repeated. The two supernatants were pooled and centrifuged 30 minutes at 30000 g and 4° C. and the pellet resuspended with a needle and syringe in a SPG buffer (3 ml/Plate). After a brief sonication the suspension was gently layered over a 30% Diatrizoate solution (50 g Meglumine diatrizoate, 7.7 g Sodium diatrizoate in 76 ml H₂O) and centrifuged at 40,000 g for 30 min. After centrifugation the pellet were resuspended in SPG buffer and stored at -70° C. The IFU of the batches were quantified by titration on McCoy cells and the concentration of the batches was determined by BCA.

Antigen and Fusion Preparation Methods

[0194] The genome of C. trachomatis serovar D, E, F and G are publicly available (NCBI-GenBank). Genes coding for C. trachomatis antigens and fusions where all obtained synthetically for cloning into E. coli bacterial protein expression system (DNA2.0). The pET411 vector was used for expression of the recombinant C. trachomatis protein in E. coli with a Histidine affinity tag. The bacterial host was BL21-START®. E. coli was grown at 37° C. to reach the logarithmic phase OD600˜0.5 and protein expression was induced for 4 hours and cells were harvested by centrifugation (6,000 g for 15 min.). E. coli were lysed using Bugbuster (Novagen) containing Benzonase, rLysozyme and Protease inhibitor Cocktail I (Calbiochem). Inclusion bodies were isolated by centrifugation (10,000 g for 10 min.) The pellet was dissolved in 50 mM NaH2PO4, 0.4M NaCl, 8M Urea, 10 mM Imidazole pH 7.5 and loaded onto HisTrap HP column (Amersham Biosciences) and bound proteins were eluted by applying a gradient of 50 to 500 mM imidazole. Depending on the antigen and fusions isoelectric point they were further purified by ion exchange chromatography. Protein concentrations was determined by BCA protein assay (Pierce).

Animals

[0195] Female B6C3F1 mice, 8-12 weeks of age, were obtained from Harlan Laboratories. Animals were housed under standard environmental conditions and provided standard food and water ad libitum. The use of mice is guided by the regulations set forward by the Danish Ministry of Justice (Lov om dyreforsog, jvf lovbekendelser nr. 726 of 9. September 1993), and Animal protection committees. A detailed description of the experiments was submitted to and approved by the regional ethical review board (2012-15-2934-00100) held by the applicant.

Immunization

[0196] Mice were immunized 3 times with 14 days between immunizations. The poly peptides were emulsified in CAF01 and administered simultaneously by the subcutanous (sc) and intranasal (i.n) route. The vaccines given by both routes consisted of 5 ug of peptide (see above) emulsified in 250 ug DDA and 100 ug TDB. As a negative control, DDA/TDB alone, without peptide was injected.

Chlamydia-Specific Cellular Responses

[0197] Blood lymfocytes or splenocytes were purified. Blood lymphocytes were pooled from 8 mice in each group and spenocytes were cultivated individually (n=4) and cultured in triplicate in round-bottomed microtiter plates (Nunc, Denmark) containing 2×10⁵ cells/well in a volumen of 200 μl RPMI-1640 supplemented with 5×10^-5M 2-mercaptoethanol, 1 mM glutamine, 1% pyruvate, 1% penicillin-streptomycin, 1% HEPES and 10% fetal calf serum (FCS) (Invitrogen, Denmark). The cells were re-stimulated with individual antigens in 1-10 μg/ml or VD1 and VD4 peptide pools (2 μg/ml of each peptide). Stimulation with Concanavalin A (5 μg/ml) or media as positive control for cell viability and negative control, respectively. After 72 h of incubation at 37° C. in 5% CO₂, supernatants were harvested and stored at -20° C. before use. The amounts of secreted IFN-γ were determined by enzyme-linked immunosorbant assay (ELISA).

Serum Antibodies

[0198] At different time points post last vaccination the mice were bled and serum isolated by centrifugation. Serum was tested by ELISA for reactivity against the Ct surface (SvD, SvE and SvF), against the SvE VD4 monomer, and against peptides (Table 4&5) spanning the VD4 region of SvD, SvE and SvF. Briefly, plates were coated with antigen (1 to 10 ug/ml) at 4° C. in carbonate buffer overnight, blocked with BSA and washed. The plates were then incubated with pre-diluted samples at 4° C. overnight, washed and incubated with a peroxidase conjugated secondary antibody for 1 hr. Reactions were visualized by incubation with TMB substrate and the reaction stopped with sulphuric acid and read at 450 nm. When ELISA reactivity against a 9mer overlapping peptide panel spanning the VD4 region of SvD (SvE) (Table 6) and SvF (Table 7) was investigated minor changes were done. Briefly, plates were treated with streptavidin and coated with biotinylated peptides, blocked for 2 h at room temperature with skimmed-milk powder and washed. The plates were then incubated with pre-diluted (1:100) serum samples for 2 h at room temperature, washed and incubated with a peroxidase conjugated secondary antibody for 1 hr. Reactions were visualized by incubation with TMB substrate and the reaction stopped with sulphuric acid and read at 450 nm.

Neutralization Assay

[0199] HaK cells were grown to confluence in 96-well flat-bottom microtiter plates in RPMI 1640 media supplemented with 5% fetal calf serum (Gibco BRL; heat inactivated), 1% v/v Hepes, 1% v/v L-glutamine, 1% v/v pyrovate and 10 μg/ml gentamycine.

[0200] The Chlamydia stocks were previously titrated and diluted to 3×10⁶ IFU/ml for SvE, 2×10⁶ IFU/ml for SvD and 5×10⁶ IFU/ml for SvF. Serum (pooled) isolated from vaccinated mice was heat inactivated at 56° C. for 1/2 h, diluted 2-4 times and 4-5 fold titrated. 80 μl of the bacteria suspension was mixed with 80 μl of serum (+/-20 μg/ml peptide) and incubated for 30 min. at 3TC on a slowly rocking platform and 50 μl of the suspension were then inoculated onto the previously prepared HaK cells in duplicates. To do this, the media was removed from the HaK monolayers and 100 μl of the above media supplemented with 0.5% glucose and 10 μg/ml cyclohexamide was added followed by 50 μl of the serum/bacteria suspension. Plates were incubated at 35° C. on a slowly rocking platform, then inoculum was removed and 100 μl of the above media supplemented with 0.5% glucose and 10 μg/ml cycloheximide was added. The plates were then incubated for 24 h at 3TC in an atmosphere of 5% CO₂ in humidified air. After incubation the medium was removed and the monolayers were fixed with 96% ethanol for 10 min. Inclusions were visualized by staining with polyclonal rabbit anti-CT755 serum made in our laboratory, followed by FITC-conjugated swine anti-rabbit immunoglobulin (Dako). Background staining was done with propidium iodide (Invitrogen)

Vaginal Challenge and Vaginal Chlamydial Load

[0201] Ten and 3 days before Ct serovar D challenge, the oestrus cycle was synchronized by injection of 2.5 mg Medroxyprogesteronacetat (Depo-Provera; Pfizer). Six weeks after the final vaccination the mice were challenged i.vag. with 4-8×10⁵ IFU of Ct serovar D in 10 μl SPG buffer. Vaginal swabs were obtained at 3, 7, 10 and 14 days after infection. Swabs were vortexed with glass-beads in 0.6 ml SPG buffer and stored at -80 C until analysis. Infectious load was determined as described in¹⁷. Briefly, McCoy cell monolayers were infected with a titrated volume of the swab suspension in duplicates. The plates were centrifuged at 750×g for 1 h at RT followed by incubation at 35 C for 2 h. Infection-media was then replaced with fresh media and the cells incubated at 37 C for 30 h. Inclusions were visualised by staining with polyclonal rabbit anti-CT681 serum made in our laboratory, followed by a FITC conjugated swine anti-rabbit Ig (DAKO, Glostrup, Denmark). Background staining was done with propidium iodide (Invitrogen, Taastrup, Denmark). Inclusions were enumerated by fluorescence microscopy observing at least 20 individual fields of vision for each well.

Depletion of CD4.sup.+ and CD8.sup.+ T-cells

[0202] Monoclonal anti-mouse CD4 (clone GK1.5) and anti-mouse CD8 (clone YTS156 and YTS169 a gift from Stephen Cobbold)⁷⁸, 79 was purified from hybridoma supernatants made in our lab, using HiTrap protein G HP columns (GE-Healthcare Life Sciences, Denmark). The purified IgG was dialyzed against PBS, filtered through 0.22 um filter and protein concentration was determined by OD 280 nm. Mice were depleted of CD4.sup.+ or CD8.sup.+ T-cells by 4 injections of 250-300 μg purified anti-CD4 or a mix of anti-CD8 antibodies at day-7, -4, -1 and +2 and +6 relative to the day of infection. The CD4.sup.+ and CD8.sup.+ T cell depletions were verified by FACS analysis on PBMCs at day 1 post infection using a FITC conjugated anti-CD4 antibody (clone RM4-4) and a PE-conjugated anti-CD8 antibody (clone 53-6) (BD Biosciences, Denmark).

In Vivo Depletion

[0203] The Chlamydia serovar D stock was previously titrated and diluted to 8×10⁴ IFU/μl, mixed 1:1 with serum isolated from mice immunized with a heterologous VD4 immuno-repeat SvD-SvE-SvF (CTH89). Ten and 3 days before Ct serovar D challenge, the oestrus cycle was synchronized by injection of 2.5 mg Medroxyprogesteronacetat (Depo-Provera; Pfizer). Mice were challenged i.vag. with 10 μl of the above mix (4×10⁵ IFU of Ct serovar D). Vaginal swabs were obtained at 3, 7 and 10 days after infection.

Statistical Analysis

[0204] Statistical analysis was done using GraphPad Prism 4. Medians of vaginal Chlamydia load were analyzed using Kruskall-Wallis followed by Dunn's post test or Mann-Whitney.

Example 1

[0205] Enhanced immune responses after immunization with homologous immuno-repeats of VD4^ext compared with a monomeric VD^ext unit.

Introduction

[0206] Here we selected polypeptide units containing extended VD4 fragments of serovar E (for sequence see FIG. 2) (SvE VD4^ext). In order to potentiate the immune response against these domains we designed recombinant polypeptides were the SvEVD4^ext unit was presented in a repetitive manner. To investigate if a repetitive form of the construct could enhance the antibody response compared to a monomeric form, we designed recombinant polypeptides where the units were presented either as a single unit or in a repetetive manner. For serovar E (SvE), a monomeric (SvE VD4^ext)*1 (CTH181), four immuno-repeats (SvE VD4^ext)*4 (CTH527) and eight immuno-repeats (SvE VD4^ext)*8 (CTH526) of the extended VD4 unit were constructed. These homologous immuno-repeat constructs were formulated in the adjuvant CAF01 and used to vaccinate mice; each mice was vaccinated with 2×5 μg peptide so the amount of VD4 was the same. Immunogenicity of the constructs was studied by ELISA against SvE VD4^ext, peptides covering SvE VD4^ext and the bacterial surface of chlamydia.

Results

[0207] Six mice/group were immunized 2 times with 14 days between immunizations. The vaccines (2×5 μg) were emulsified in CAF01 and administered simultaneously by the sc. and i.n routes. At certain time points post last vaccination blood was collected and antibody levels against the extended VD4 units from SvE and against the bacterial surface of SvE were measured by ELISA. Vaccination with a single VD4^ext unit (monomeric VD4^ext, CTH181) induced lower levels of VD4^ext specific antibodies compared to the level induced after immunization with homologous immuno-repeats composed of 4 VD4^ext repeats of (SvE VD4^ext)*4 (FIG. 5A). The higher antibody response seen after immunizing with (SvE VD4^ext).sup.*4 resulted in a stronger recognition of the bacterial surface compared to serum isolated from (VD4^ext).sup.*1 immunized mice (FIG. 5B). The response to 20mer peptides with 10aa overlap spanning the extended VD4 region (Table 4) was also enhanced resulting in a broader epitope recognition pattern in the (VD4^ext)*4 homologous immuno-repeat groups compared to the group of mice immunized with a monomeric VD4^ext unit when tested in a 1:500 serum dilution (FIG. 5C). In the group immunized with the monomeric construct the response was exclusively targeted to the central region containing the TTLNPTIAG epitope whereas immunization with the homologous immuno-repeat exposed several B cell epitopes both up- and downstream of that epitope resulting in a diverse epitope recognition pattern of various epitopes. We continued by investigating if immuno-repeats of 8 (SvE VD4^ext)*8 (CTH526, seq no 30) were more immunogenic than immuno-repeats of 4 (SvE VD4^ext)*4. The two constructs induced similar levels of antibodies against the extended VD4 unit and against the bacterial surface of SvE.

Conclusion

[0208] We demonstrated that by immunizing with immuno-repeats of extended VD4 units from Serovar E we can greatly enhance antibody response both measured as the titer (FIGS. 5A&B) and the breadth of the response (FIG. 5C) directed against the extended VD4 unit resulting in a strong reactivity towards the bacterial surface. We did not find enhanced antibody titers and neutralization titers by increasing the number of repeats from 4 to 8.

Example 2

[0209] A construct composed of heterologous immuno-repeats from SvD, E, F and G (CTH518) induced a stronger response to multiple serovars compared to homologous immuno-repeats from SvF

Introduction

[0210] We investigated if immunization with at heterologous immuno-repeat composed of extended VD4 units from SvD, SvE, SvF and SvG (CTH518), maintained the strong immunogenicity and was able to induce a broader antibody response recognizing the surface of multiple serovars compared to immunization with a homologous immuno-repeat composed of extended VD4 units from SvF (SvF VD4^extt)*4, (CTH529). These immuno-repeat constructs were formulated in the adjuvant CAF01 and used to vaccinate mice. The immunogenicity of the constructs was studied by ELISA against the bacterial surface of Serovar D, E and F.

Results

[0211] Heterologous immuno-repeats promoted an antibody response that recognized the surface of the serovar F strain at the same high level as the response seen with a homologous immuno-repeat from SvF. However, by immunization with the heterologous immuno-repeat containing extended VD4 regions from the four serotypes (SvD, SvE, SvF, SvG) we observed a markedly increased titer to the D and E serovariants compared to the homologous immuno-repeat from the serovar F (FIG. 6).

Conclusion

[0212] Immunizing with the construct composed of immuno-repeats of heterologous extended VD4's induced a broader response recognizing the surface of multiple serovars (D, E and F) while maintaining the pronounced immunogenicity of the homologous immuno-repeat.

Example 3

[0213] The specificity of the antibody responses after immunization with a heterologous immuno-repeat of the extended VD4 units from serovar D, E, and F (CTH89) compared to constructs composed of a homologous immuno-repeat from (SvE^ext VD4)*4, (SvF^ext. VD4)*4 and a previously published A8-VD4 peptide⁶⁵.

Introduction

[0214] We investigated the specificity of the immune response after immunization with a heterologous repeat of extended VD4 domains from SvD, SvE, SvF (CTH89) compared to immunization with homologous immuno-repeats composed of extended VD4 repeats from Serovar E (SvE^extVD4)*4 (CTH527), SvF (SvF^extVD4)*4 repeats (CTH524) and A8-VD4 peptide. These constructs were formulated in the adjuvant CAF01 and used to vaccinate mice. Immunogenicity of the constructs was studied by ELISA against a peptide panel (9 and 20 AA long) spanning the VD4 region of D, E and F (Tables 4-7). Serum (from 6 to 8 mice) was tested and a response above background but below OD=1.0 is indicated by an open box, responses above 1.0 are marked by a filled box. The length of the box indicates the area recognized by antibodies.

Results

[0215] All constructs induced high antibody responses to the conserved TTLNPTIAG (SEQ ID NO 76) part of the VD4^ext, located in the variable domain (VD). In general antibodies generated by homologous immuno-repeats were superior in recognizing their representative homologous VD4^ext region, whereas it was evident that when these constructs were tested against peptides covering a VD4^ext from a different serovar their epitope recognition repertoire was limited e.g. the recognition of serovar E VD4 region by serum from animals immunized with the construct (SvF^extVD4)*4 (FIGS. 7A and C) (and vice versa) (FIGS. 7B and C).

[0216] Antibodies generated after immunization with the heterologous immuno-repeats (CTH89), recognized a much broader epitope repertoire than serum from animals immunized with the homologous immuno-repeats and the A8-VD4 (7A, B, C and D). This construct was able to cover an epitope repertoire covering both serovar E and F at the level (or better) than achieved by immunizing with homologous immuno-repeats.

[0217] To demonstrate whether a 17 AA peptide representing a central VD4 peptide FDTTTLNPTIAGAGDVK (SEQ ID NO 194) was able to compete with C. trachomatis organisms for CTH89 specific antibody binding, a competitive neutralization assay was performed. Different concentrations of CTH89 and A8-VD4 specific serum were mixed with the peptide in a concentration of 20 μg/ml (FIG. 7D). The results demonstrates that, in contrast to A8-VD4 specific serum, the peptide could not completely eliminate the neutralizing capacity of the CTH89 specific serum, suggesting that this serum targets a broader repertoire of neutralizing epitopes.

Conclusion

[0218] Immunizing with immuno-repeats of heterologous extended VD4's induced a broad response recognizing both conserved and serovar specific parts of the VD4 region, translating into a broader repertoire of neutralizing epitopes.

Example 4

[0219] Immunization with heterologous immuno-repeats of extended VD4's from SvD, SvE and SvF (CTH89) generates early T cell independent protection after a SvD challenge.

Introduction

[0220] In order to study the effector mechanism responsible for the early protection seen after vaccination with the VD4 repetitive units, mice vaccinated with CTH89 were T cell depleted before challenge and the capacity to induce early protection was compared in depleted and non-depleted mice.

Results

[0221] Eight mice/group were immunized 3 times with 14 days between immunizations. The vaccine (2×5 μg) was emulsified in CAF01 and administered simultaneously by the sc. and i.n routes. At certain time points post last vaccination the mice were bleed and antibody responses against chlamydia, the neutralization titer, and in vivo protection with and without T cell depletion were measured. Depletion of the T cell subset eliminated the T cell response to CTH89 (FIG. 8A). CTH89 induced a strong antibody response (FIG. 8B) that recognized the surface of serovar D (FIG. 8C) and was able to neutralize the bacteria in vitro with a 50% neutralization titer of around 1:10³ (FIG. 8D). However, we still found significant protection at day 3 post challenge in the T cell depleted mice (FIG. 8E) suggesting an in vivo role for antibodies recognizing the VD4 unit in early protection against Chlamydia. Finally we demonstrated that CTH89 serum was also able to neutralize a SvE and SvF infection with very high 50% neutralization titers at the level of that obtained with SvD (FIG. 8D).

Conclusions

[0222] Immuno-repeat generates T cell independent early protection against vaginal challenge with Serovar D suggesting an in vivo role of VD4 specific antibodies.

Example 5

In Vivo Neutralization with CTH89 Specific Serum

Introduction

[0223] In order to investigate if the in vitro neutralization could be translated to a protective effect mediated by serum in vivo, we next investigated if SvD bacteria coated with antibodies generated after CTH89 immunization could neutralize/inhibit the infection in vivo compared to serum from naive mice.

Results

[0224] SvD bacteria were mixed with serum isolated from CTH89 immunized mice or serum isolated from naive mice. Depro-provera treated mice were then infected with 4×10⁵ bacteria. Mice infected with SvD coated with CTH89 serum efficiently controlled bacterial replication compared to mice challenged with SvD coated with naive serum. Six out of 8 mice were cleared at day 7 and 10 compared to 2 and 3 respectively, in the control group (FIG. 9).

Conclusion

[0225] Serum generated after immunization with heterologous VD4 immuno-repeat efficiently block infection of mice with SvD compared to serum isolated from naive mice

Example 6

Fusion of Recombinant MOMP with Immuno-Repeats of Heterologous Extended VD4's

Introduction

[0226] MOMP is the target of both humoral and cellular immune-responses but despite the relative success of refolded native MOMP vaccines in generating neutralizing antibodies and protect against infection⁵⁴, 56, experimental vaccines based on recombinant MOMP (rMOMP) have failed. We designed a recombinant MOMP ranging from amino acid 56 to 349, including all variable domains (CTH521). We also selected polypeptide units containing extended VD4 fragments (covering the VD4 variable domain of MOMP and the adjacent conserved flanking regions) of serovar D, E, F and G (CT518) Finally a hybrid was constructed where CTH521 was fused to CTH518 (CT522) (FIG. 10).

Results

[0227] Eight mice/group were immunized 3 times with 14 days between immunizations. The vaccines were emulsified in CAF01 and administered simultaneously by the sc. (5 μg) and i.n. (5 μg) routes. Post vaccination blood samples were collected and antibodies against the VD4^ext unit, recombinant MOMP and against the bacterial surface were measured. Antibodies generated after immunization with CT522 and CT518 recognized the VD4 region (FIG. 10A) and the bacterial surface (FIG. 10C) at a much higher level compared to serum isolated after CT521 immunization. Furthermore antibodies form CTH518 and CTH522 were able to neutralize a SvD infection at the same level and much higher than CTH521 (FIG. 10D).

Conclusion

[0228] Fusion of recombinant MOMP with immuno-repeats of heterologous extended VD4's results in a molecule that elicits the same functional antibody response as the immune-repeat alone.

Example 7

[0229] Vaccination with heterologous immuno-repeats of VD1^ext-VD4^ext's regions from SvD, SvE and SvF (CTH88) compared to vaccination with a single VD1-VD4 unit from SvD (CTH87).

Introduction

[0230] We next investigated if it was possible to fuse another VD region to the extended VD4 region and still maintain the capacity to induce neutralizing antibodies. Therefore constructs were designed were an extended version of the VD1 region was coupled to the extended VD4 region. We produced both a homologous unit composed of an extended unit of VD1 and VD4 from SvD (CTH87) and a heterologous immuno-repeat composed of extended units of VD1 and VD4 from different serovars (D, E and F; CTH88).

Results

[0231] 12 mice/group were immunized 3 times with 14 days between immunizations. The vaccines were emulsified in CAF01 and administered simultaneously by the sc. (5 μg) and i.n. (5 μg) routes Antibodies from mice immunized with CTH87 recognized the bacterial surface of both SvD, SvE and SvF (FIG. 11A); with the highest titers observed against the homologous SvD strain and the lowest titers against the most distant SvF. Immunizing with immuno-repeats of heterologous VD1^ext-VD4^ext units resulted in significant higher levels of antibodies against the surface of the bacteria compared to the monomeric construct and broadened the response resulting in titers increasing 6-12 times against SvD and SvE and almost 25 times against SvF (FIG. 11A). The capacity of these antibodies to neutralize infection in an in vitro neutralizing assay was even more improved as serum from animals immunized with the monomeric VD1^ext-VD4^ext construct from serovar D only had minimal neutralizing capacity compared to the heterologous VD1-VD4 immuno-repeat construct with a neutralization titer of 1:2000 (FIG. 11B). Finally did vaccination with the heterologous VD1^ext-VD4^ext immuno-repeat construct very efficiently protect against a SvD challenge in a vaginal challenge model (FIG. 11C).

Conclusion

[0232] We demonstrated that by immunizing with immuno-repeats of heterologous VD1^ext-VD4^ext units from serovar D, E and F, we can greatly enhance the antibody response directed against the bacterial surface of all three serovariants. Importantly we also show that by vaccination with a heterologous immuno-repeat, we observe a selective higher increase in Serovar F surface recognition (25 times vs. 6-12 times for serovar D and E), suggesting that the heterologous immuno-repeats not only increase the antibody levels against shared epitopes but also against serovar F specific epitopes. We demonstrated that the antibodies induced with immuno-repeats of heterologous VD1-VD4 (CTH88) generated in vitro neutralizing titers that resulted in early in vivo protection compared to the single VD1-VD4 unit from SvD (CTH87) (FIG. 11C).

Example 8

Coupling of T Cell Antigens to Immuno-Repeats of VD4

Introduction

[0233] As there is a generally recognized need for a CMI component in an efficient protective immune response against Chlamydia trachomatis, we next investigated if the heterologous immuno-repeats can be fused to T cell antigens with vaccine potential. Our aim was to provide both an early antibody mediated protection against Ct as well as an efficient CMI mediated clearance of residual organisms. A constructs composed of CT043, and part of CT414 and CT681 was fused to immuno-repeats of heterologous VD1-VD4 (CTH91).

Results

[0234] 12 mice/group were immunized 3 times with 14 days between immunizations. The vaccines (2×5 μg) were emulsified in CAF01 and administered by the sc. and i.n. routes. At various time points post last vaccination the mice were bleed and antibody responses and neutralization titers were measured. Antibodies generated after immunization with CTH91 and CTH88 recognized the VD4^ext region at similar levels (FIG. 12A) and serum isolated from both groups were able to neutralize a SvD infection (FIG. 12B). Compared to CTH88 immunized mice the T cell response to CTH91 was stronger with recognition of both CT414 and CT043 (FIG. 12C). This T and B cell response resulted in significant protection at day 3 post infection for both groups, but at day 7 and 10 post infection the group vaccinated with a fused T and B cell target (CTH91) induced higher levels of protection compared to CTH88 (FIG. 12D).

Conclusion

[0235] We were able to fuse T cell antigens with the repetitive VD regions and still maintain the capacity to induce early protection and moreover these constructs induced an efficient CMI mediated clearance of residual organisms leading to high levels of protection at day 7 post infection.

Example 9

Immunization with a Cocktail of a Heterologous VD4 Immuno-Repeat and a T Cell Antigen Fusion Molecule

Introduction

[0236] We next investigated if immuno-repeats can be mixed with T cell antigens with vaccine potential and still provide both an early antibody mediated protection against Ct as well as an efficient CMI mediated clearance of residual organisms. We therefore investigated if we could mix a strong T cell hybrid composed of CT043, part of CT414 and CT681 (CTH93) with CTH89 (FIG. 13A) and still maintain the capacity to neutralize the SvD bacteria in vitro and induce early protection against a vaginal challenge.

Results

[0237] 12 mice/group were immunized 3 times with 14 days between immunizations. The vaccine (2×5 μg) were emulsified in CAF01 and administered simultaneously by the subcutaneous (sc) and intranasal (i.n) route (FIG. 13). Antibodies generated after immunization with CTH89 or the mixture of CTH89 and CTH93 strongly recognized the VD4 regions (FIG. 13B) and neutralized the bacteria with similar 50% neutralization titers (FIG. 13C). Much reduced levels of VD4 recognition and neutralization was seen after vaccination with the T cell antigen fusion (CTH93, FIG. 13D) although this molecules also contained MOMP (CT681) and therefore potentially the same neutralizing epitopes. This molecule also gave very low levels of recognition of the TTLNPTIAG epitope (data not shown). This clearly emphasizes the limitation of full-size recombinant MOMP as a vaccine antigen for the induction of neutralizing antibodies as previously reported. Both the CTH89 and the cocktail of the CTH89 and CTH93 vaccines induced protection at day 3 post infection (FIG. 13E). This was in contrast to CTH93 vaccinated mice which induced no significant protection at day 3 post infection. At day 7 post infection both vaccines including the strong T cell target (CTH93) induced a significant level of protection (FIG. 13D&E).

Conclusions

[0238] We were able to mix the heterologous VD4 repeats with strong T cell antigens without the loss of in vitro neutralization and early in vivo protection against a Serovar D challenge. Moreover, the mix of B and T cell targets induced an efficient CMI mediated clearance of residual organisms leading to high levels of protection at day 7 post infection.

Example 10

Testing the Effect of Different Adjuvant Systems

Introduction

[0239] In order to investigate if the high antibody response against heterologous immuno-repeats were only seen when the vaccine were administered in CAF01- we compared the antibody response and the neutralization titer after immunizing with CTH527 (SvE VD4^ext)*4 in CAF01 or Alum.

Results

[0240] Both adjuvant systems induced a high antibody response against the surface of SvE when administered together with CTH527, and the antibodies from both groups were able to neutralize SvE in vitro (FIG. 14).

Example 11

Vaccination with Heterologous Immuno-Repeats Composed of Reduced Length of the VD4^ext regions from SvD, SvE, SvF and SvG

Introduction

[0241] We next compared heterologous immuno-repeat constructs composed of reduced length of the VD4 region (CTH285 (SEQ ID NO 69) and CTH286 (SEQ ID NO 70)) compared to the CTH518 construct (CTH518 (SEQ ID NO 53)) (FIG. 15A).

Results

[0242] 4 mice/group were immunized 3 times with 14 days between immunizations. The vaccines were emulsified in CAF01 and administered simultaneously by the subcutaneous (sc, 5 μg) and intranasal (i.n, 5 μg) routes. Splenocytes from 4 mice/group were isolated and the T cell responses to overlapping peptides representing the VD4^ext region (FIG. 15B) and the capacity of the serum to neutralize a serovar D and F infection (FIG. 15C) were investigated. Much reduced levels of VD4 T cell recognition, and neutralization was seen after vaccination with CTH285 where the VD4^ext regions from the different serovars were reduced with 38 aa. CTH286 on the other hand (each VD4^ext region reduced with 24 aa) induced similar levels of T cell responses and had the same capacity to neutralize a serovar D infection as CTH518.

Conclusion

[0243] We demonstrated that by reducing the length of the VD4^ext regions with 38 aa we reduced both the T cell responses and the capacity to neutralize a serovar D and F infection.

Example 12

[0244] Vaccination with heterologous immuno-repeats composed of extended VD4^ext regions from SvD, SvE, SvF, SvG, SvIa and SvJ.

Introduction

[0245] We next investigated if we by extending the length of the VD4^ext region could enhance the T cell response to the immuno-repeat constructs. We designed two constructs CTH69 (SEQ ID NO 47) and CTH72 (SEQ ID NO 48). CTH69 was similar to CTH88 but the VD4^ext regions from SvD, SvE and SvF was extended by 12aa N-terminally (FIG. 16B). CTH72 also contained VD1 and VD4^ext regions from SvG, SvIa and SvJ.

Results

[0246] Mice were immunized 3 times with 14 days between immunizations. The vaccines were emulsified in CAF01 and administered simultaneously by the subcutaneous (sc, 5 μg) and intranasal (i.n, 5 μg) routes. T cell responses to the antigen used for immunization and to peptide pools representing the VD1 and VD4 regions from the different serovars were investigated (FIG. 16). Extending the VD4^ext regions induced a significant higher T cell response (>40.000 μg/ml) compared to the T cell response obtained with CTH88 (<20.000 pg/ml) (FIG. 16B). Importantly, both of the extended constructs were still able to neutralize a serovar D infection in vitro (FIG. 16C). Comparing the protective efficacy of the vaccines, CTH69 and CTH72 induced a significant level of protection at day 7 post infection which could possibly be explained by the stronger T cell response induced by these vaccines compared to CTH88 (FIG. 16D).

Conclusion

[0247] Extending the VD4^ext region enhanced the T cell response compared to CTH88 which led to enhanced protection at day 7 post infection.

REFERENCES

[0248] 1. WHO. Global Prevalence and Incidence of selected Curable Sexually Transmitted Infections: Overview and Estimates. World Health Organization, Geneva, Switzerland; 2001.

[0249] 2. Paavonen J, Eggert-Kruse W. Chlamydia trachomatis: impact on human reproduction. Hum Reprod Update 1999, 5(5): 433-447.

[0250] 3. Plummer F A, Simonsen J N, Cameron D W, Ndinya-Achola J O, Kreiss J K, Gakinya M N, et al. Cofactors in male-female sexual transmission of human immunodeficiency virus type 1. J Infect Dis 1991, 163(2): 233-239.

[0251] 4. Anttila T, Saikku P, Koskela P, Bloigu A, Dillner J, Ikaheimo I, et al. Serotypes of Chlamydia trachomatis and risk for development of cervical squamous cell carcinoma. Jama 2001, 285(1): 47-51.

[0252] 5. Golden M R, Schillinger J A, Markowitz L, St Louis M E. Duration of untreated genital infections with chlamydia trachomatis: a review of the literature. Sex Transm Dis 2000, 27(6): 329-337.

[0253] 6. Batteiger B E, Xu F, Johnson R E, Rekart M L. Protective immunity to Chlamydia trachomatis genital infection: evidence from human studies. J Infect Dis, 201 Suppl 2: S178-189.

[0254] 7. Brunham R C, Rey-Ladino J. Immunology of Chlamydia infection: implications for a Chlamydia trachomatis vaccine. Nat Rev Immunol 2005, 5(2): 149-161.

[0255] 8. Su H, Caldwell H D. CD4+ T cells play a significant role in adoptive immunity to Chlamydia trachomatis infection of the mouse genital tract. Infect Immun 1995, 63(9): 3302-3308.

[0256] 9. Morrison S G, Su H, Caldwell H D, Morrison R P. Immunity to murine Chlamydia trachomatis genital tract reinfection involves B cells and CD4(+) T cells but not CD8(+) T cells. Infect Immun 2000, 68(12): 6979-6987.

[0257] 10. Morrison R P, Caldwell H D. Immunity to murine chlamydial genital infection. Infect Immun 2002, 70(6): 2741-2751.

[0258] 11. Rasmussen S J. Chlamydia immunology. Curr Opin Infect Dis 1998, 11(1): 37-41.

[0259] 12. Rank R. In: Chlamydia Intracellular Biology, Pathogenesis and Immunity. Washington D.C. ASM Press 1999: Pp. 239-296.

[0260] 13. Morrison S G, Morrison R P. Resolution of secondary Chlamydia trachomatis genital tract infection in immune mice with depletion of both CD4+ and CD8+ T cells. Infect Immun 2001, 69(4): 2643-2649.

[0261] 14. Moore T, Ekworomadu C O, Eko F O, MacMillan L, Ramey K, Ananaba G A, et al. Fc receptor-mediated antibody regulation of T cell immunity against intracellular pathogens. J Infect Dis 2003, 188(4): 617-624.

[0262] 15. Pal S, Rangel J, Peterson E M, de la Maza L M. Immunogenic and protective ability of the two developmental forms of Chlamydiae in a mouse model of infertility. Vaccine 1999, 18(7-8): 752-761.

[0263] 16. Darville T, Hiltke T J. Pathogenesis of genital tract disease due to Chlamydia trachomatis. J Infect Dis 2010, 201 Suppl 2: 5114-125.

[0264] 17. Hansen J, Jensen K T, Follmann F, Agger E M, Theisen M, Andersen P. Liposome Delivery of Chlamydia muridarum Major Outer Membrane Protein Primes a Th1 Response That Protects against Genital Chlamydial Infection in a Mouse Model. J Infect Dis 2008, 198(5): 758-767.

[0265] 18. Olsen A W, Theisen M, Christensen D, Follmann F, Andersen P. Protection against Chlamydia promoted by a subunit vaccine (CTH1) compared with a primary intranasal infection in a mouse genital challenge model. PLoS One, 5(5): e10768.

[0266] 19. Li W, Murthy A K, Guentzel M N, Chambers J P, Forsthuber T G, Seshu J, et al. Immunization with a combination of integral chlamydial antigens and a defined secreted protein induces robust immunity against genital chlamydial challenge. Infect Immun 2010, 78(9): 3942-3949.

[0267] 20. Olsen A W, Follmann F, Hojrup P, Leah R, Sand C, Andersen P, et al. Identification of human T-cell targets recognized during the Chlamydia trachomatis genital infection. J Infect Dis 2007, 196: 1546-1552.

[0268] 21. Olsen A W, Follmann F, Jensen K, Hojrup P, Leah R, Sorensen H, et al. Identification of CT521 as a frequent target of Th1 cells in patients with urogenital Chlamydia trachomatis infection. J Infect Dis 2006, 194(9): 1258-1266.

[0269] 22. Follmann F, Olsen A W, Jensen K T, Hansen P R, Andersen P, Theisen M. Antigenic profiling of a Chlamydia trachomatis gene-expression library. J Infect Dis 2008, 197 897-905.

[0270] 23. Sharma J, Zhong Y, Dong F, Piper J M, Wang G, Zhong G. Profiling of human antibody responses to Chlamydia trachomatis urogenital tract infection using microplates arrayed with 156 chlamydial fusion proteins. Infect Immun 2006, 74(3): 1490-1499.

[0271] 24. Coler R N, Bhatia A, Maisonneuve J F, Probst P, Barth B, Ovendale P, et al. Identification and characterization of novel recombinant vaccine antigens for immunization against genital Chlamydia trachomatis. FEMS Immunol Med Microbiol 2009, 55(2): 258-270.

[0272] 25. Karunakaran K P, Rey-Ladino J, Stoynov N, Berg K, Shen C, Jiang X, et al. Immunoproteomic discovery of novel T cell antigens from the obligate intracellular pathogen Chlamydia. J Immunol 2008, 180(4): 2459-2465.

[0273] 26. Yu H, Jiang X, Shen C, Karunakaran K P, Brunham R C. Novel Chlamydia muridarum T cell antigens induce protective immunity against lung and genital tract infection in murine models. J Immunol 2009, 182(3): 1602-1608.

[0274] 27. Molina D M, Pal S, Kayala M A, Teng A, Kim P J, Baldi P, et al. Identification of immunodominant antigens of Chlamydia trachomatis using proteome microarrays. Vaccine 2010, 28(17): 3014-3024.

[0275] 28. Stephens R S, Kalman S, Lammel C, Fan J, Marathe R, Aravind L, et al. Genome sequence of an obligate intracellular pathogen of humans: Chlamydia trachomatis. Science 1998, 282(5389): 754-759.

[0276] 29. Sette A, Rappuoli R. Reverse vaccinology: developing vaccines in the era of genomics. Immunity 2010, 33(4): 530-541.

[0277] 30. Igietseme J U, Eko F O, Black C M. Chlamydia vaccines: recent developments and the role of adjuvants in future formulations. Expert Rev Vaccines 2011, 10(11): 1585-1596.

[0278] 31. Rockey D D, Wang J, Lei L, Zhong G. Chlamydia vaccine candidates and tools for chlamydial antigen discovery. Expert Rev Vaccines 2009, 8(10): 1365-1377.

[0279] 32. Farris C M, Morrison R P. Vaccination against Chlamydia genital infection utilizing the murine C. muridarum model. Infect Immun 2011, 79(3): 986-996.

[0280] 33. Kubo A, Stephens R S. Characterization and functional analysis of PorB, a Chlamydia porin and neutralizing target. Mol Microbiol 2000, 38(4): 772-780.

[0281] 34. Kawa D E, Schachter J, Stephens R S. Immune response to the Chlamydia trachomatis outer membrane protein PorB. Vaccine 2004, 22(31-32): 4282-4286.

[0282] 35. Crane D D, Carlson J H, Fischer E R, Bavoil P, Hsia R C, Tan C, et al. Chlamydia trachomatis polymorphic membrane protein D is a species-common pan-neutralizing antigen. Proc Natl Acad Sci U SA 2006, 103(6): 1894-1899.

[0283] 36. Baehr W, Zhang Y X, Joseph T, Su H, Nano F E, Everett K D, et al. Mapping antigenic domains expressed by Chlamydia trachomatis major outer membrane protein genes. Proc Natl Acad Sci USA 1988, 85(11): 4000-4004.

[0284] 37. Bavoil P, Ohlin A, Schachter J. Role of disulfide bonding in outer membrane structure and permeability in Chlamydia trachomatis. Infect Immun 1984, 44(2): 479-485.

[0285] 38. Hatch T P, Allan I, Pearce J H. Structural and polypeptide differences between envelopes of infective and reproductive life cycle forms of Chlamydia spp. J Bacteriol 1984, 157(1): 13-20.

[0286] 39. Stephens R S, Sanchez-Pescador R, Wagar E A, Inouye C, Urdea M S. Diversity of Chlamydia trachomatis major outer membrane protein genes. J Bacteriol 1987, 169(9): 3879-3885.

[0287] 40. Caldwell H D, Perry L J. Neutralization of Chlamydia trachomatis infectivity with antibodies to the major outer membrane protein. Infect Immun 1982, 38(2): 745-754.

[0288] 41. Peeling R, Maclean I W, Branham R C. In vitro neutralization of Chlamydia trachomatis with monoclonal antibody to an epitope on the major outer membrane protein. Infect Immun 1984, 46(2): 484-488.

[0289] 42. Zhang Y X, Stewart S, Joseph T, Taylor H R, Caldwell H D. Protective monoclonal antibodies recognize epitopes located on the major outer membrane protein of Chlamydia trachomatis. J Immunol 1987, 138(2): 575-581.

[0290] 43. Zhang Y X, Stewart S J, Caldwell H D. Protective monoclonal antibodies to Chlamydia trachomatis serovar- and serogroup-specific major outer membrane protein determinants. Infect Immun 1989, 57(2): 636-638.

[0291] 44. Cotter T W, Meng Q, Shen Z L, Zhang Y X, Su H, Caldwell H D. Protective efficacy of major outer membrane protein-specific immunoglobulin A (IgA) and IgG monoclonal antibodies in a murine model of Chlamydia trachomatis genital tract infection. Infect Immun 1995, 63(12): 4704-4714.

[0292] 45. Bandea C I, Debattista J, Joseph K, Igietseme J, Timms P, Black C M. Chlamydia trachomatis serovars among strains isolated from members of rural indigenous communities and urban populations in Australia. J Clin Microbiol 2008, 46(1): 355-356.

[0293] 46. Hsu M C, Tsai P Y, Chen K T, Li L H, Chiang C C, Tsai J J, et al. Genotyping of Chlamydia trachomatis from clinical specimens in Taiwan. J Med Microbiol 2006, 55(Pt 3): 301-308.

[0294] 47. Jonsdottir K, Kristjansson M, Hjaltalin Olafsson J, Steingrimsson O. The molecular epidemiology of genital Chlamydia trachomatis in the greater Reykjavik area, Iceland. Sex Transm Dis 2003, 30(3): 249-256.

[0295] 48. Lysen M, Osterlund A, Rubin C J, Persson T, Persson I, Herrmann B. Characterization of ompA genotypes by sequence analysis of DNA from all detected cases of Chlamydia trachomatis infections during 1 year of contact tracing in a Swedish County. J Clin Microbiol 2004, 42(4): 1641-1647.

[0296] 49. Millman K, Black C M, Johnson R E, Stamm W E, Jones R B, Hook E W, et al. Population-based genetic and evolutionary analysis of Chlamydia trachomatis urogenital strain variation in the United States. J Bacteriol 2004, 186(8): 2457-2465.

[0297] 50. Millman K, Black C M, Stamm W E, Jones R B, Hook E W, 3rd, Martin D H, et al. Population-based genetic epidemiologic analysis of Chlamydia trachomatis serotypes and lack of association between ompA polymorphisms and clinical phenotypes. Microbes Infect 2006, 8(3): 604-611.

[0298] 51. Su H, Parnell M, Caldwell H D. Protective efficacy of a parenterally administered MOMP-derived synthetic oligopeptide vaccine in a murine model of Chlamydia trachomatis genital tract infection: serum neutralizing IgG antibodies do not protect against chlamydial genital tract infection. Vaccine 1995, 13(11): 1023-1032.

[0299] 52. Pal S, Barnhart K M, Wei Q, Abai A M, Peterson E M, de la Maza L M. Vaccination of mice with DNA plasmids coding for the Chlamydia trachomatis major outer membrane protein elicits an immune response but fails to protect against a genital challenge. Vaccine 1999, 17(5): 459-465.

[0300] 53. Zhang D J, Yang X, Shen C, Brunham R C. Characterization of immune responses following intramuscular DNA immunization with the MOMP gene of Chlamydia trachomatis mouse pneumonitis strain. Immunology 1999, 96(2): 314-321.

[0301] 54. Pal S, Theodor I, Peterson E M, de la Maza L M Immunization with the Chlamydia trachomatis mouse pneumonitis major outer membrane protein can elicit a protective immune response against a genital challenge. Infect Immun 2001, 69(10): 6240-6247.

[0302] 55. Shaw J, Grund V, Durling L, Crane D, Caldwell H D. Dendritic cells pulsed with a recombinant chlamydial major outer membrane protein antigen elicit a CD4(+) type 2 rather than type 1 immune response that is not protective. Infect Immun 2002, 70(3): 1097-1105.

[0303] 56. Kari L, Whitmire W M, Crane D D, Reveneau N, Carlson J H, Goheen M M, et al. Chlamydia trachomatis native major outer membrane protein induces partial protection in nonhuman primates: implication for a trachoma transmission-blocking vaccine. J Immunol 2009, 182(12): 8063-8070.

[0304] 57. Carmichael J R, Pal S, Tifrea D, de la Maza L M. Induction of protection against vaginal shedding and infertility by a recombinant Chlamydia vaccine. Vaccine 2011, 29(32): 5276-5283.

[0305] 58. Yen T Y, Pal S, de la Maza L M. Characterization of the disulfide bonds and free cysteine residues of the Chlamydia trachomatis mouse pneumonitis major outer membrane protein. Biochemistry 2005, 44(16): 6250-6256.

[0306] 59. Stephens R S, Wagar E A, Schoolnik G K. High-resolution mapping of serovar-specific and common antigenic determinants of the major outer membrane protein of Chlamydia trachomatis. J Exp Med 1988, 167(3): 817-831.

[0307] 60. Murdin A D, Su H, Klein M H, Caldwell H D. Poliovirus hybrids expressing neutralization epitopes from variable domains I and IV of the major outer membrane protein of Chlamydia trachomatis elicit broadly cross-reactive C. trachomatis-neutralizing antibodies. Infect Immun 1995, 63(3): 1116-1121.

[0308] 61. Murdin A D, Su H, Manning D S, Klein M H, Parnell M J, Caldwell H D. A poliovirus hybrid expressing a neutralization epitope from the major outer membrane protein of Chlamydia trachomatis is highly immunogenic. Infect Immun 1993, 61(10): 4406-4414.

[0309] 62. Villeneuve A, Brossay L, Paradis G, Hebert J. Determination of neutralizing epitopes in variable domains I and IV of the major outer-membrane protein from Chlamydia trachomatis serovar K. Microbiology 1994, 140 (Pt 9): 2481-2487.

[0310] 63. Villeneuve A, Brossay L, Paradis G, Hebert J. Characterization of the humoral response induced by a synthetic peptide of the major outer membrane protein of Chlamydia trachomatis serovar B. Infect Immun 1994, 62(8): 3547-3549.

[0311] 64. Motin V L, de la Maza L M, Peterson E M. Immunization with a peptide corresponding to chlamydial heat shock protein 60 increases the humoral immune response in C3H mice to a peptide representing variable domain 4 of the major outer membrane protein of Chlamydia trachomatis. Clin Diagn Lab Immunol 1999, 6(3): 356-363.

[0312] 65. Su H, Caldwell H D. Immunogenicity of a synthetic oligopeptide corresponding to antigenically common T-helper and B-cell neutralizing epitopes of the major outer membrane protein of Chlamydia trachomatis. Vaccine 1993, 11(11): 1159-1166.

[0313] 66. Toye B, Zhong G M, Peeling R, Brunham R C Immunologic characterization of a cloned fragment containing the species-specific epitope from the major outer membrane protein of Chlamydia trachomatis. Infect Immun 1990, 58(12): 3909-3913.

[0314] 67. Mygind P, Christiansen G, Persson K, Birkelund S. Detection of Chlamydia trachomatis-specific antibodies in human sera by recombinant major outer-membrane protein polyantigens. J Med Microbiol 2000, 49(5): 457-465.

[0315] 68. Qu Z, Cheng X, de la Maza L M, Peterson E M. Analysis of the humoral response elicited in mice by a chimeric peptide representing variable segments I and IV of the major outer membrane protein of Chlamydia trachomatis. Vaccine 1994, 12(6): 557-564.

[0316] 69. Peterson E M, Cheng X, Qu Z, de la Maza L M. The effect of orientation within a chimeric peptide on the immunogenicity of Chlamydia trachomatis

epitopes. Mol Immunol 1996, 33(4-5): 335-339.

[0317] 70. Caldwell H D, Kromhout J, Schachter J. Purification and partial characterization of the major outer membrane protein of Chlamydia trachomatis. Infect Immun 1981, 31(3): 1161-1176.

[0318] 71. Ravn P, Demissie A, Eguale T, Wondwosson H, Lein D, Amoudy H A, et al. Human T cell responses to the ESAT-6 antigen from Mycobacterium tuberculosis. J Infect Dis 1999, 179(3): 637-645.

[0319] 72. Stryhn A, Pedersen L O, Romme T, Holm C B, Holm A, Buus S. Peptide binding specificity of major histocompatibility complex class I resolved into an array of apparently independent subspecificities: quantitation by peptide libraries and improved prediction of binding. Eur J Immunol 1996, 26(8): 1911-1918.

[0320] 73. Harboe M, Oettinger T, Wiker H G, Rosenkrands I, Andersen P. Evidence for occurrence of the ESAT-6 protein in Mycobacterium tuberculosis and virulent Mycobacterium bovis and for its absence in Mycobacterium bovis BCG. Infect Immun 1996, 64(1): 16-22.

[0321] 74. Volp K, Mathews S, Timms P, Hafner L. Peptide immunization of guinea pigs against Chlamydia psittaci (GPIC agent) infection induces good vaginal secretion antibody response, in vitro neutralization and partial protection against live challenge. Immunol Cell Biol 2001, 79(3): 245-250.

[0322] 75. Hinton H J, Jegerlehner A, Bachmann M F. Pattern recognition by B cells: the role of antigen repetitiveness versus Toll-like receptors. Current topics in microbiology and immunology 2008, 319: 1-15.

[0323] 76. Kim S K, DeMars R. Epitope clusters in the major outer membrane protein of Chlamydia trachomatis. Curr Opin Immunol 2001, 13(4): 429-436.

[0324] 77. Findlay H E, McClafferty H, Ashley R H. Surface expression, single-channel analysis and membrane topology of recombinant Chlamydia trachomatis Major Outer Membrane Protein. BMC Microbiol 2005, 5: 5.

[0325] 78. Cobbold S P, Jayasuriya A, Nash A, Prospero T D, Waldmann H. Therapy with monoclonal antibodies by elimination of T-cell subsets in vivo. Nature 1984, 312(5994): 548-551.

[0326] 79. Qin S, Cobbold S, Tighe H, Benjamin R, Waldmann H. CD4 monoclonal antibody pairs for immunosuppression and tolerance induction. Eur J Immunol 1987, 17(8): 1159-1165.

Sequence CWU 1

1

255120PRTChlamydia trachomatis 1Ala Lys Pro Thr Thr Asp Thr Gly Asn Ser Ala Ala Pro Ser Thr Leu 1 5 10 15 Thr Ala Arg Glu 20 220PRTChlamydia trachomatis 2Asp Lys Pro Thr Ser Thr Thr Gly Asn Ala Thr Ala Pro Thr Thr Leu 1 5 10 15 Thr Ala Arg Glu 20 321PRTChlamydia trachomatis 3Glu Ala Leu Ala Gly Ala Ser Gly Asn Thr Thr Ser Thr Leu Ser Lys 1 5 10 15 Leu Val Glu Arg Thr 20 421PRTChlamydia trachomatis 4Glu Ala Leu Ala Gly Ala Ser Gly Asn Thr Thr Ser Thr Leu Ser Lys 1 5 10 15 Leu Val Glu Arg Thr 20 522PRTChlamydia trachomatis 5Ala Ala Pro Thr Thr Lys Asp Ile Ala Gly Leu Glu Asn Asp Pro Thr 1 5 10 15 Thr Asn Val Ala Arg Pro 20 622PRTChlamydia trachomatis 6Ala Ala Pro Thr Thr Ser Asp Val Ala Gly Leu Gln Asn Asp Pro Thr 1 5 10 15 Thr Asn Val Ala Arg Pro 20 730PRTChlamydia trachomatis 7Asp Ala Ile Ser Met Arg Val Gly Tyr Tyr Gly Asp Phe Val Phe Asp 1 5 10 15 Arg Val Leu Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly 20 25 30 818PRTChlamydia trachomatis 8Asn Pro Ala Tyr Gly Arg His Met Gln Asp Ala Glu Met Phe Thr Asn 1 5 10 15 Ala Ala 959PRTChlamydia trachomatis 9Ala Ile Ser Met Arg Val Gly Tyr Tyr Gly Asp Phe Val Phe Asp Arg 1 5 10 15 Val Leu Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly Ala Lys Pro 20 25 30 Thr Thr Asp Thr Gly Asn Ser Ala Ala Pro Ser Thr Leu Thr Ala Arg 35 40 45 Glu Asn Pro Ala Tyr Gly Arg His Met Gln Asp 50 55 1059PRTChlamydia trachomatis 10Ala Ile Ser Met Arg Met Gly Tyr Tyr Gly Asp Phe Val Phe Asp Arg 1 5 10 15 Val Leu Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly Asp Lys Pro 20 25 30 Thr Ser Thr Thr Gly Asn Ala Thr Ala Pro Thr Thr Leu Thr Ala Arg 35 40 45 Glu Asn Pro Ala Tyr Gly Arg His Met Gln Asp 50 55 1160PRTChlamydia trachomatis 11Ala Ile Ser Met Arg Met Gly Tyr Tyr Gly Asp Phe Val Phe Asp Arg 1 5 10 15 Val Leu Lys Thr Asp Val Asn Lys Glu Phe Glu Met Gly Glu Ala Leu 20 25 30 Ala Gly Ala Ser Gly Asn Thr Thr Ser Thr Leu Ser Lys Leu Val Glu 35 40 45 Arg Thr Asn Pro Ala Tyr Gly Lys His Met Gln Asp 50 55 60 1260PRTChlamydia trachomatis 12Ala Ile Ser Met Arg Met Gly Tyr Tyr Gly Asp Phe Val Phe Asp Arg 1 5 10 15 Val Leu Lys Thr Asp Val Asn Lys Glu Phe Glu Met Gly Glu Ala Leu 20 25 30 Ala Gly Ala Ser Gly Asn Thr Thr Ser Thr Leu Ser Lys Leu Val Glu 35 40 45 Arg Thr Asn Pro Ala Tyr Gly Lys His Met Gln Asp 50 55 60 1361PRTChlamydia trachomatis 13Ala Ile Ser Met Arg Met Gly Tyr Tyr Gly Asp Phe Val Phe Asp Arg 1 5 10 15 Val Leu Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly Ala Ala Pro 20 25 30 Thr Thr Lys Asp Ile Ala Gly Leu Glu Asn Asp Pro Thr Thr Asn Val 35 40 45 Ala Arg Pro Asn Pro Ala Tyr Gly Lys His Met Gln Asp 50 55 60 1461PRTChlamydia trachomatis 14Ala Ile Ser Met Arg Met Gly Tyr Tyr Gly Asp Phe Val Phe Asp Arg 1 5 10 15 Val Leu Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly Ala Ala Pro 20 25 30 Thr Thr Ser Asp Val Ala Gly Leu Gln Asn Asp Pro Thr Thr Asn Val 35 40 45 Ala Arg Pro Asn Pro Ala Tyr Gly Lys His Met Gln Asp 50 55 60 1530PRTChlamydia trachomatis 15Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala 1 5 10 15 Gly Ala Gly Asp Val Lys Thr Gly Ala Glu Gly Gln Leu Gly 20 25 30 1630PRTChlamydia trachomatis 16Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala 1 5 10 15 Gly Ala Gly Asp Val Lys Ala Ser Ala Glu Gly Gln Leu Gly 20 25 30 1731PRTChlamydia trachomatis 17Leu Val Thr Pro Val Val Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala 1 5 10 15 Gly Cys Gly Ser Val Ala Gly Ala Asn Thr Glu Gly Gln Ile Ser 20 25 30 1831PRTChlamydia trachomatis 18Leu Ala Lys Pro Val Val Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala 1 5 10 15 Gly Cys Gly Ser Val Val Ala Ala Asn Ser Glu Gly Gln Ile Ser 20 25 30 1931PRTChlamydia trachomatis 19Leu Ala Glu Ala Ile Leu Asp Val Thr Thr Leu Asn Pro Thr Ile Ala 1 5 10 15 Gly Lys Gly Thr Val Val Ala Ser Gly Ser Asp Asn Asp Leu Ala 20 25 30 2031PRTChlamydia trachomatis 20Leu Ala Glu Ala Ile Leu Asp Val Thr Thr Leu Asn Pro Thr Ile Ala 1 5 10 15 Gly Lys Gly Thr Val Val Ala Ser Gly Ser Glu Asn Asp Leu Ala 20 25 30 2139PRTChlamydia trachomatis 21Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr Arg Leu Asn Met Phe Thr 1 5 10 15 Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr 20 25 30 Ile Arg Ile Ala Gln Pro Lys 35 2230PRTChlamydia trachomatis 22Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Lys Met Lys Ser Arg 1 5 10 15 Lys Ser Cys Gly Ile Ala Val Gly Thr Thr Ile Val Asp Ala 20 25 30 2368PRTChlamydia trachomatis 23Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe 1 5 10 15 Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile 20 25 30 Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val 35 40 45 Lys Thr Gly Ala Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser 50 55 60 Leu Gln Leu Asn 65 2468PRTChlamydia trachomatis 24Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe 1 5 10 15 Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile 20 25 30 Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val 35 40 45 Lys Ala Ser Ala Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser 50 55 60 Leu Gln Leu Asn 65 2569PRTChlamydia trachomatis 25Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe 1 5 10 15 Asp Ser Asp Thr Ile Arg Ile Ala Gln Pro Arg Leu Val Thr Pro Val 20 25 30 Val Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala Gly Cys Gly Ser Val 35 40 45 Ala Gly Ala Asn Thr Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val 50 55 60 Ser Leu Gln Leu Asn 65 2669PRTChlamydia trachomatis 26Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe 1 5 10 15 Asp Ser Asn Thr Ile Arg Ile Ala Gln Pro Lys Leu Ala Lys Pro Val 20 25 30 Val Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala Gly Cys Gly Ser Val 35 40 45 Val Ala Ala Asn Ser Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val 50 55 60 Ser Leu Gln Leu Asn 65 2769PRTChlamydia trachomatis 27Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Val Ser Phe 1 5 10 15 Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Leu Ala Glu Ala Ile 20 25 30 Leu Asp Val Thr Thr Leu Asn Pro Thr Ile Ala Gly Lys Gly Thr Val 35 40 45 Val Ala Ser Gly Ser Asp Asn Asp Leu Ala Asp Thr Met Gln Ile Val 50 55 60 Ser Leu Gln Leu Asn 65 2869PRTChlamydia trachomatis 28Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Val Ser Phe 1 5 10 15 Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Leu Ala Glu Ala Ile 20 25 30 Leu Asp Val Thr Thr Leu Asn Pro Thr Ile Ala Gly Lys Gly Thr Val 35 40 45 Val Ala Ser Gly Ser Glu Asn Asp Leu Ala Asp Thr Met Gln Ile Val 50 55 60 Ser Leu Gln Leu Asn 65 2921PRTChlamydia trachomatis 29Asp Asn Glu Asn Gln Lys Thr Val Lys Ala Glu Ser Val Pro Asn Met 1 5 10 15 Ser Phe Asp Gln Ser 20 3021PRTChlamydia trachomatis 30Asp Asn Glu Asn Gln Ser Thr Val Lys Thr Asn Ser Val Pro Asn Met 1 5 10 15 Ser Leu Asp Gln Ser 20 3121PRTChlamydia trachomatis 31Asp Gly Val Asn Ala Thr Lys Pro Ala Ala Asp Ser Ile Pro Asn Val 1 5 10 15 Gln Leu Asn Gln Ser 20 3221PRTChlamydia trachomatis 32Asp Ser Glu Asn Ala Thr Gln Pro Ala Ala Thr Ser Ile Pro Asn Val 1 5 10 15 Gln Leu Asn Gln Ser 20 3322PRTChlamydia trachomatis 33Thr Lys Thr Gln Ser Ser Asn Phe Asn Thr Ala Lys Leu Ile Pro Asn 1 5 10 15 Ala Ala Leu Asn Gln Ala 20 3422PRTChlamydia trachomatis 34Thr Lys Thr Gln Ala Ser Ser Phe Asn Thr Ala Asn Leu Phe Pro Asn 1 5 10 15 Thr Ala Leu Asn Gln Ala 20 3523PRTChlamydia trachomatis 35Thr Leu Gly Ala Thr Ser Gly Tyr Leu Lys Gly Asn Ser Ala Ser Phe 1 5 10 15 Asn Leu Val Gly Leu Phe Gly 20 3622PRTChlamydia trachomatis 36Val Val Glu Leu Tyr Thr Asp Thr Thr Phe Ala Trp Ser Val Gly Ala 1 5 10 15 Arg Ala Ala Leu Trp Glu 20 3714PRTChlamydia trachomatis 37Lys Glu Phe Pro Leu Asp Leu Thr Ala Gly Thr Asp Ala Ala 1 5 10 3814PRTChlamydia trachomatis 38Gln Glu Phe Pro Leu Ala Leu Ile Ala Gly Thr Asp Ala Ala 1 5 10 3914PRTChlamydia trachomatis 39Lys Glu Phe Pro Leu Asp Leu Thr Ala Gly Thr Asp Ala Ala 1 5 10 4014PRTChlamydia trachomatis 40Gln Glu Phe Pro Leu Ala Leu Thr Ala Gly Thr Asp Ala Ala 1 5 10 4114PRTChlamydia trachomatis 41Ala Glu Phe Pro Leu Asp Ile Thr Ala Gly Thr Glu Ala Ala 1 5 10 4214PRTChlamydia trachomatis 42Ala Glu Phe Pro Leu Asp Ile Thr Ala Gly Thr Glu Ala Ala 1 5 10 4338PRTChlamydia trachomatis 43Ala Thr Leu Gly Ala Ser Phe Gln Tyr Ala Gln Ser Lys Pro Lys Val 1 5 10 15 Glu Glu Leu Asn Val Leu Cys Asn Ala Ala Glu Phe Thr Ile Asn Lys 20 25 30 Pro Lys Gly Tyr Val Gly 35 4435PRTChlamydia trachomatis 44Thr Gly Thr Lys Asp Ala Ser Ile Asp Tyr His Glu Trp Gln Ala Ser 1 5 10 15 Leu Ala Leu Ser Tyr Arg Leu Asn Met Phe Thr Pro Tyr Ile Gly Val 20 25 30 Lys Trp Ser 35 45127PRTChlamydia trachomatis 45Ala Ile Ser Met Arg Val Gly Tyr Tyr Gly Asp Phe Val Phe Asp Arg 1 5 10 15 Val Leu Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly Ala Lys Pro 20 25 30 Thr Thr Asp Thr Gly Asn Ser Ala Ala Pro Ser Thr Leu Thr Ala Arg 35 40 45 Glu Asn Pro Ala Tyr Gly Arg His Met Gln Asp Asn Met Phe Thr Pro 50 55 60 Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr Ile 65 70 75 80 Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr 85 90 95 Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Thr Gly Ala Glu 100 105 110 Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 115 120 125 46383PRTChlamydia trachomatis 46Ala Ile Ser Met Arg Val Gly Tyr Tyr Gly Asp Phe Val Phe Asp Arg 1 5 10 15 Val Leu Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly Ala Lys Pro 20 25 30 Thr Thr Asp Thr Gly Asn Ser Ala Ala Pro Ser Thr Leu Thr Ala Arg 35 40 45 Glu Asn Pro Ala Tyr Gly Arg His Met Gln Asp Asn Met Phe Thr Pro 50 55 60 Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr Ile 65 70 75 80 Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr 85 90 95 Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Thr Gly Ala Glu 100 105 110 Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Ala 115 120 125 Ile Ser Met Arg Met Gly Tyr Tyr Gly Asp Phe Val Phe Asp Arg Val 130 135 140 Leu Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly Asp Lys Pro Thr 145 150 155 160 Ser Thr Thr Gly Asn Ala Thr Ala Pro Thr Thr Leu Thr Ala Arg Glu 165 170 175 Asn Pro Ala Tyr Gly Arg His Met Gln Asp Asn Met Phe Thr Pro Tyr 180 185 190 Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg 195 200 205 Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu 210 215 220 Asn Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Ala Ser Ala Glu Gly 225 230 235 240 Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Ala Ile 245 250 255 Ser Met Arg Met Gly Tyr Tyr Gly Asp Phe Val Phe Asp Arg Val Leu 260 265 270 Lys Thr Asp Val Asn Lys Glu Phe Glu Met Gly Glu Ala Leu Ala Gly 275 280 285 Ala Ser Gly Asn Thr Thr Ser Thr Leu Ser Lys Leu Val Glu Arg Thr 290 295 300 Asn Pro Ala Tyr Gly Lys His Met Gln Asp Asn Met Phe Thr Pro Tyr 305 310 315 320 Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ser Asp Thr Ile Arg 325 330 335 Ile Ala Gln Pro Arg Leu Val Thr Pro Val Val Asp Ile Thr Thr Leu 340 345 350 Asn Pro Thr Ile Ala Gly Ser Gly Ser Val Ala Gly Ala Asn Thr Glu 355 360 365 Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 370 375 380 47419PRTChlamydia trachomatis 47Ala Ile Ser Met Arg Val Gly Tyr Tyr Gly Asp Phe Val Phe Asp Arg 1 5 10 15 Val Leu Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly Ala Lys Pro 20 25 30 Thr Thr Asp Thr Gly Asn Ser Ala Ala Pro Ser Thr Leu Thr Ala Arg 35 40 45 Glu Asn Pro Ala Tyr Gly Arg His Met Gln Asp Glu Trp Gln Ala Ser 50 55 60 Leu Ala Leu Ser Tyr Arg

Leu Asn Met Phe Thr Pro Tyr Ile Gly Val 65 70 75 80 Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln 85 90 95 Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr 100 105 110 Ile Ala Gly Ala Gly Asp Val Lys Thr Gly Ala Glu Gly Gln Leu Gly 115 120 125 Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Ala Ile Ser Met Arg 130 135 140 Met Gly Tyr Tyr Gly Asp Phe Val Phe Asp Arg Val Leu Lys Thr Asp 145 150 155 160 Val Asn Lys Glu Phe Gln Met Gly Asp Lys Pro Thr Ser Thr Thr Gly 165 170 175 Asn Ala Thr Ala Pro Thr Thr Leu Thr Ala Arg Glu Asn Pro Ala Tyr 180 185 190 Gly Arg His Met Gln Asp Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr 195 200 205 Arg Leu Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala 210 215 220 Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr 225 230 235 240 Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly 245 250 255 Asp Val Lys Ala Ser Ala Glu Gly Gln Leu Gly Asp Thr Met Gln Ile 260 265 270 Val Ser Leu Gln Leu Asn Ala Ile Ser Met Arg Met Gly Tyr Tyr Gly 275 280 285 Asp Phe Val Phe Asp Arg Val Leu Lys Thr Asp Val Asn Lys Glu Phe 290 295 300 Glu Met Gly Glu Ala Leu Ala Gly Ala Ser Gly Asn Thr Thr Ser Thr 305 310 315 320 Leu Ser Lys Leu Val Glu Arg Thr Asn Pro Ala Tyr Gly Lys His Met 325 330 335 Gln Asp Glu Trp Gln Ala Ser Leu Ser Leu Ser Tyr Arg Leu Asn Met 340 345 350 Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ser 355 360 365 Asp Thr Ile Arg Ile Ala Gln Pro Arg Leu Val Thr Pro Val Val Asp 370 375 380 Ile Thr Thr Leu Asn Pro Thr Ile Ala Gly Ser Gly Ser Val Ala Gly 385 390 395 400 Ala Asn Thr Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu 405 410 415 Gln Leu Asn 48844PRTChlamydia trachomatis 48Ala Ile Ser Met Arg Val Gly Tyr Tyr Gly Asp Phe Val Phe Asp Arg 1 5 10 15 Val Leu Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly Ala Lys Pro 20 25 30 Thr Thr Asp Thr Gly Asn Ser Ala Ala Pro Ser Thr Leu Thr Ala Arg 35 40 45 Glu Asn Pro Ala Tyr Gly Arg His Met Gln Asp Glu Trp Gln Ala Ser 50 55 60 Leu Ala Leu Ser Tyr Arg Leu Asn Met Phe Thr Pro Tyr Ile Gly Val 65 70 75 80 Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln 85 90 95 Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr 100 105 110 Ile Ala Gly Ala Gly Asp Val Lys Thr Gly Ala Glu Gly Gln Leu Gly 115 120 125 Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Ala Ile Ser Met Arg 130 135 140 Met Gly Tyr Tyr Gly Asp Phe Val Phe Asp Arg Val Leu Lys Thr Asp 145 150 155 160 Val Asn Lys Glu Phe Gln Met Gly Asp Lys Pro Thr Ser Thr Thr Gly 165 170 175 Asn Ala Thr Ala Pro Thr Thr Leu Thr Ala Arg Glu Asn Pro Ala Tyr 180 185 190 Gly Arg His Met Gln Asp Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr 195 200 205 Arg Leu Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala 210 215 220 Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr 225 230 235 240 Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly 245 250 255 Asp Val Lys Ala Ser Ala Glu Gly Gln Leu Gly Asp Thr Met Gln Ile 260 265 270 Val Ser Leu Gln Leu Asn Ala Ile Ser Met Arg Met Gly Tyr Tyr Gly 275 280 285 Asp Phe Val Phe Asp Arg Val Leu Lys Thr Asp Val Asn Lys Glu Phe 290 295 300 Glu Met Gly Glu Ala Leu Ala Gly Ala Ser Gly Asn Thr Thr Ser Thr 305 310 315 320 Leu Ser Lys Leu Val Glu Arg Thr Asn Pro Ala Tyr Gly Lys His Met 325 330 335 Gln Asp Glu Trp Gln Ala Ser Leu Ser Leu Ser Tyr Arg Leu Asn Met 340 345 350 Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ser 355 360 365 Asp Thr Ile Arg Ile Ala Gln Pro Arg Leu Val Thr Pro Val Val Asp 370 375 380 Ile Thr Thr Leu Asn Pro Thr Ile Ala Gly Ser Gly Ser Val Ala Gly 385 390 395 400 Ala Asn Thr Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu 405 410 415 Gln Leu Asn Ala Ile Ser Met Arg Met Gly Tyr Tyr Gly Asp Phe Val 420 425 430 Phe Asp Arg Val Leu Lys Thr Asp Val Asn Lys Glu Phe Glu Met Gly 435 440 445 Glu Ala Leu Ala Gly Ala Ser Gly Asn Thr Thr Ser Thr Leu Ser Lys 450 455 460 Leu Val Glu Arg Thr Asn Pro Ala Tyr Gly Lys His Met Gln Asp Glu 465 470 475 480 Trp Gln Ala Ser Leu Ser Leu Ser Tyr Arg Leu Asn Met Phe Thr Pro 485 490 495 Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ser Asn Thr Ile 500 505 510 Arg Ile Ala Gln Pro Lys Leu Ala Lys Pro Val Val Asp Ile Thr Thr 515 520 525 Leu Asn Pro Thr Ile Ala Gly Ser Gly Ser Val Val Ala Ala Asn Ser 530 535 540 Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 545 550 555 560 Ala Ile Ser Met Arg Met Gly Tyr Tyr Gly Asp Phe Val Phe Asp Arg 565 570 575 Val Leu Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly Ala Ala Pro 580 585 590 Thr Thr Lys Asp Ile Ala Gly Leu Glu Asn Asp Pro Thr Thr Asn Val 595 600 605 Ala Arg Pro Asn Pro Ala Tyr Gly Lys His Met Gln Asp Glu Trp Gln 610 615 620 Ala Ser Leu Ala Leu Ser Tyr Arg Leu Asn Met Phe Thr Pro Tyr Ile 625 630 635 640 Gly Val Lys Trp Ser Arg Val Ser Phe Asp Ala Asp Thr Ile Arg Ile 645 650 655 Ala Gln Pro Lys Leu Ala Glu Ala Ile Leu Asp Val Thr Thr Leu Asn 660 665 670 Pro Thr Ile Ala Gly Lys Gly Thr Val Val Ala Ser Gly Ser Asp Asn 675 680 685 Asp Leu Ala Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Ala Ile 690 695 700 Ser Met Arg Met Gly Tyr Tyr Gly Asp Phe Val Phe Asp Arg Val Leu 705 710 715 720 Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly Ala Ala Pro Thr Thr 725 730 735 Ser Asp Val Ala Gly Leu Gln Asn Asp Pro Thr Thr Asn Val Ala Arg 740 745 750 Pro Asn Pro Ala Tyr Gly Lys His Met Gln Asp Glu Trp Gln Ala Ser 755 760 765 Leu Ala Leu Ser Tyr Arg Leu Asn Met Phe Thr Pro Tyr Ile Gly Val 770 775 780 Lys Trp Ser Arg Val Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln 785 790 795 800 Pro Lys Leu Ala Glu Ala Ile Leu Asp Val Thr Thr Leu Asn Pro Thr 805 810 815 Ile Ala Gly Lys Gly Thr Val Val Ala Ser Gly Ser Glu Asn Asp Leu 820 825 830 Ala Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 835 840 49205PRTChlamydia trachomatis 49Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe 1 5 10 15 Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile 20 25 30 Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val 35 40 45 Lys Thr Gly Ala Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser 50 55 60 Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser 65 70 75 80 Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser 85 90 95 Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly 100 105 110 Ala Gly Asp Val Lys Ala Ser Ala Glu Gly Gln Leu Gly Asp Thr Met 115 120 125 Gln Ile Val Ser Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr Ile Gly 130 135 140 Val Lys Trp Ser Arg Ala Ser Phe Asp Ser Asp Thr Ile Arg Ile Ala 145 150 155 160 Gln Pro Arg Leu Val Thr Pro Val Val Asp Ile Thr Thr Leu Asn Pro 165 170 175 Thr Ile Ala Gly Ser Gly Ser Val Ala Gly Ala Asn Thr Glu Gly Gln 180 185 190 Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 195 200 205 5075PRTChlamydia trachomatis 50Met His His His His His His Asn Met Phe Thr Pro Tyr Ile Gly Val 1 5 10 15 Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln 20 25 30 Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr 35 40 45 Ile Ala Gly Ala Gly Asp Val Lys Ala Ser Ala Glu Gly Gln Leu Gly 50 55 60 Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 65 70 75 5169PRTChlamydia trachomatis 51Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe 1 5 10 15 Asp Ser Asp Thr Ile Arg Ile Ala Gln Pro Arg Leu Val Thr Pro Val 20 25 30 Val Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala Gly Cys Gly Ser Val 35 40 45 Ala Gly Ala Asn Thr Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val 50 55 60 Ser Leu Gln Leu Asn 65 5276PRTChlamydia trachomatis 52Met His His His His His His Asn Met Phe Thr Pro Tyr Ile Gly Val 1 5 10 15 Lys Trp Ser Arg Ala Ser Phe Asp Ser Asp Thr Ile Arg Ile Ala Gln 20 25 30 Pro Arg Leu Val Thr Pro Val Val Asp Ile Thr Thr Leu Asn Pro Thr 35 40 45 Ile Ala Gly Cys Gly Ser Val Ala Gly Ala Asn Thr Glu Gly Gln Ile 50 55 60 Ser Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 65 70 75 53274PRTChlamydia trachomatis 53Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe 1 5 10 15 Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile 20 25 30 Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val 35 40 45 Lys Thr Gly Ala Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser 50 55 60 Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser 65 70 75 80 Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser 85 90 95 Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly 100 105 110 Ala Gly Asp Val Lys Ala Ser Ala Glu Gly Gln Leu Gly Asp Thr Met 115 120 125 Gln Ile Val Ser Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr Ile Gly 130 135 140 Val Lys Trp Ser Arg Ala Ser Phe Asp Ser Asp Thr Ile Arg Ile Ala 145 150 155 160 Gln Pro Arg Leu Val Thr Pro Val Val Asp Ile Thr Thr Leu Asn Pro 165 170 175 Thr Ile Ala Gly Ser Gly Ser Val Ala Gly Ala Asn Thr Glu Gly Gln 180 185 190 Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Asn Met Phe 195 200 205 Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ser Asn 210 215 220 Thr Ile Arg Ile Ala Gln Pro Lys Leu Ala Lys Pro Val Val Asp Ile 225 230 235 240 Thr Thr Leu Asn Pro Thr Ile Ala Gly Ser Gly Ser Val Val Ala Ala 245 250 255 Asn Ser Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln 260 265 270 Leu Asn 54322PRTChlamydia trachomatis 54Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr Arg Leu Asn Met Phe Thr 1 5 10 15 Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr 20 25 30 Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr 35 40 45 Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Thr Gly Ala 50 55 60 Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 65 70 75 80 Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr Arg Leu Asn Met Phe Thr 85 90 95 Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr 100 105 110 Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr 115 120 125 Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Ala Ser Ala 130 135 140 Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 145 150 155 160 Glu Trp Gln Ala Ser Leu Ser Leu Ser Tyr Arg Leu Asn Met Phe Thr 165 170 175 Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ser Asp Thr 180 185 190 Ile Arg Ile Ala Gln Pro Arg Leu Val Thr Pro Val Val Asp Ile Thr 195 200 205 Thr Leu Asn Pro Thr Ile Ala Gly Ser Gly Ser Val Ala Gly Ala Asn 210 215 220 Thr Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln Leu 225 230 235 240 Asn Glu Trp Gln Ala Ser Leu Ser Leu Ser Tyr Arg Leu Asn Met Phe 245 250 255 Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ser Asn 260 265 270 Thr Ile Arg Ile Ala Gln Pro Lys Leu Ala Lys Pro Val Val Asp Ile 275 280 285 Thr Thr Leu Asn Pro Thr Ile Ala Gly Ser Gly Ser Val Val Ala Ala 290 295 300 Asn Ser Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln 305 310 315 320 Leu Asn 55484PRTChlamydia trachomatis 55Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr Arg Leu Asn Met Phe Thr 1 5 10 15 Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr 20 25 30 Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr 35 40 45 Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Thr Gly Ala 50 55

60 Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 65 70 75 80 Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr Arg Leu Asn Met Phe Thr 85 90 95 Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr 100 105 110 Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr 115 120 125 Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Ala Ser Ala 130 135 140 Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 145 150 155 160 Glu Trp Gln Ala Ser Leu Ser Leu Ser Tyr Arg Leu Asn Met Phe Thr 165 170 175 Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ser Asp Thr 180 185 190 Ile Arg Ile Ala Gln Pro Arg Leu Val Thr Pro Val Val Asp Ile Thr 195 200 205 Thr Leu Asn Pro Thr Ile Ala Gly Ser Gly Ser Val Ala Gly Ala Asn 210 215 220 Thr Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln Leu 225 230 235 240 Asn Glu Trp Gln Ala Ser Leu Ser Leu Ser Tyr Arg Leu Asn Met Phe 245 250 255 Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ser Asn 260 265 270 Thr Ile Arg Ile Ala Gln Pro Lys Leu Ala Lys Pro Val Val Asp Ile 275 280 285 Thr Thr Leu Asn Pro Thr Ile Ala Gly Ser Gly Ser Val Val Ala Ala 290 295 300 Asn Ser Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln 305 310 315 320 Leu Asn Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr Arg Leu Asn Met 325 330 335 Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Val Ser Phe Asp Ala 340 345 350 Asp Thr Ile Arg Ile Ala Gln Pro Lys Leu Ala Glu Ala Ile Leu Asp 355 360 365 Val Thr Thr Leu Asn Pro Thr Ile Ala Gly Lys Gly Thr Val Val Ala 370 375 380 Ser Gly Ser Asp Asn Asp Leu Ala Asp Thr Met Gln Ile Val Ser Leu 385 390 395 400 Gln Leu Asn Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr Arg Leu Asn 405 410 415 Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Val Ser Phe Asp 420 425 430 Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Leu Ala Glu Ala Ile Leu 435 440 445 Asp Val Thr Thr Leu Asn Pro Thr Ile Ala Gly Lys Gly Thr Val Val 450 455 460 Ala Ser Gly Ser Glu Asn Asp Leu Ala Asp Thr Met Gln Ile Val Ser 465 470 475 480 Leu Gln Leu Asn 56276PRTChlamydia trachomatis 56Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe 1 5 10 15 Asp Ser Asp Thr Ile Arg Ile Ala Gln Pro Arg Leu Val Thr Pro Val 20 25 30 Val Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala Gly Ser Gly Ser Val 35 40 45 Ala Gly Ala Asn Thr Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val 50 55 60 Ser Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp 65 70 75 80 Ser Arg Ala Ser Phe Asp Ser Asp Thr Ile Arg Ile Ala Gln Pro Arg 85 90 95 Leu Val Thr Pro Val Val Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala 100 105 110 Gly Ser Gly Ser Val Ala Gly Ala Asn Thr Glu Gly Gln Ile Ser Asp 115 120 125 Thr Met Gln Ile Val Ser Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr 130 135 140 Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ser Asp Thr Ile Arg 145 150 155 160 Ile Ala Gln Pro Arg Leu Val Thr Pro Val Val Asp Ile Thr Thr Leu 165 170 175 Asn Pro Thr Ile Ala Gly Ser Gly Ser Val Ala Gly Ala Asn Thr Glu 180 185 190 Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Asn 195 200 205 Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp 210 215 220 Ser Asp Thr Ile Arg Ile Ala Gln Pro Arg Leu Val Thr Pro Val Val 225 230 235 240 Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala Gly Ser Gly Ser Val Ala 245 250 255 Gly Ala Asn Thr Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser 260 265 270 Leu Gln Leu Asn 275 57544PRTChlamydia trachomatis 57Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe 1 5 10 15 Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile 20 25 30 Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val 35 40 45 Lys Ala Ser Ala Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser 50 55 60 Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser 65 70 75 80 Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser 85 90 95 Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly 100 105 110 Ala Gly Asp Val Lys Ala Ser Ala Glu Gly Gln Leu Gly Asp Thr Met 115 120 125 Gln Ile Val Ser Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr Ile Gly 130 135 140 Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala 145 150 155 160 Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro 165 170 175 Thr Ile Ala Gly Ala Gly Asp Val Lys Ala Ser Ala Glu Gly Gln Leu 180 185 190 Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Asn Met Phe Thr 195 200 205 Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr 210 215 220 Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr 225 230 235 240 Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Ala Ser Ala 245 250 255 Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 260 265 270 Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe 275 280 285 Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile 290 295 300 Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val 305 310 315 320 Lys Ala Ser Ala Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser 325 330 335 Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser 340 345 350 Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser 355 360 365 Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly 370 375 380 Ala Gly Asp Val Lys Ala Ser Ala Glu Gly Gln Leu Gly Asp Thr Met 385 390 395 400 Gln Ile Val Ser Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr Ile Gly 405 410 415 Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala 420 425 430 Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro 435 440 445 Thr Ile Ala Gly Ala Gly Asp Val Lys Ala Ser Ala Glu Gly Gln Leu 450 455 460 Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Asn Met Phe Thr 465 470 475 480 Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr 485 490 495 Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr 500 505 510 Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Ala Ser Ala 515 520 525 Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 530 535 540 58272PRTChlamydia trachomatis 58Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe 1 5 10 15 Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile 20 25 30 Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val 35 40 45 Lys Ala Ser Ala Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser 50 55 60 Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser 65 70 75 80 Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser 85 90 95 Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly 100 105 110 Ala Gly Asp Val Lys Ala Ser Ala Glu Gly Gln Leu Gly Asp Thr Met 115 120 125 Gln Ile Val Ser Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr Ile Gly 130 135 140 Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala 145 150 155 160 Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro 165 170 175 Thr Ile Ala Gly Ala Gly Asp Val Lys Ala Ser Ala Glu Gly Gln Leu 180 185 190 Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Asn Met Phe Thr 195 200 205 Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr 210 215 220 Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr 225 230 235 240 Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Ala Ser Ala 245 250 255 Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 260 265 270 59276PRTChlamydia trachomatis 59Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe 1 5 10 15 Asp Ser Asp Thr Ile Arg Ile Ala Gln Pro Arg Leu Val Thr Pro Val 20 25 30 Val Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala Gly Cys Gly Ser Val 35 40 45 Ala Gly Ala Asn Thr Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val 50 55 60 Ser Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp 65 70 75 80 Ser Arg Ala Ser Phe Asp Ser Asp Thr Ile Arg Ile Ala Gln Pro Arg 85 90 95 Leu Val Thr Pro Val Val Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala 100 105 110 Gly Cys Gly Ser Val Ala Gly Ala Asn Thr Glu Gly Gln Ile Ser Asp 115 120 125 Thr Met Gln Ile Val Ser Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr 130 135 140 Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ser Asp Thr Ile Arg 145 150 155 160 Ile Ala Gln Pro Arg Leu Val Thr Pro Val Val Asp Ile Thr Thr Leu 165 170 175 Asn Pro Thr Ile Ala Gly Cys Gly Ser Val Ala Gly Ala Asn Thr Glu 180 185 190 Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Asn 195 200 205 Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp 210 215 220 Ser Asp Thr Ile Arg Ile Ala Gln Pro Arg Leu Val Thr Pro Val Val 225 230 235 240 Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala Gly Cys Gly Ser Val Ala 245 250 255 Gly Ala Asn Thr Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser 260 265 270 Leu Gln Leu Asn 275 60785PRTChlamydia trachomatis 60Ser Arg Gln Asn Ala Glu Glu Asn Leu Lys Asn Phe Ala Lys Glu Leu 1 5 10 15 Lys Leu Pro Asp Val Ala Phe Asp Gln Asn Asn Thr Cys Ile Leu Phe 20 25 30 Val Asp Gly Glu Phe Ser Leu His Leu Thr Tyr Glu Glu His Ser Asp 35 40 45 Arg Leu Tyr Val Tyr Ala Pro Leu Leu Asp Gly Leu Pro Asp Asn Pro 50 55 60 Gln Arg Arg Leu Ala Leu Tyr Glu Lys Leu Leu Glu Gly Ser Met Leu 65 70 75 80 Gly Gly Gln Met Ala Gly Gly Gly Val Gly Val Ala Thr Lys Glu Gln 85 90 95 Leu Ile Leu Met His Cys Val Leu Asp Met Lys Tyr Ala Glu Thr Asn 100 105 110 Leu Leu Lys Ala Phe Ala Gln Leu Phe Ile Glu Thr Val Val Lys Trp 115 120 125 Arg Thr Val Cys Ser Asp Ile Ser Ala Gly Arg Glu Pro Thr Val Asp 130 135 140 Thr Met Pro Gln Met Pro Gln Gly Gly Gly Gly Gly Ile Gln Pro Pro 145 150 155 160 Pro Ala Gly Ile Arg Ala Thr Val Lys Ala Ile Val Glu Ser Thr Pro 165 170 175 Glu Ala Pro Glu Glu Ile Pro Pro Val Glu Gly Glu Glu Ser Thr Ala 180 185 190 Thr Glu Asp Pro Asn Ser Asn Thr Glu Gly Ser Ser Ala Asn Thr Asn 195 200 205 Leu Glu Gly Ser Gln Gly Asp Thr Ala Asp Thr Gly Thr Gly Asp Val 210 215 220 Asn Asn Glu Ser Gln Asp Thr Ser Asp Thr Gly Asn Ala Glu Ser Glu 225 230 235 240 Glu Gln Leu Gln Asp Ser Thr Gln Ser Asn Glu Glu Asn Thr Leu Pro 245 250 255 Asn Ser Asn Ile Asp Gln Ser Asn Glu Asn Thr Asp Glu Ser Ser Asp 260 265 270 Ser His Thr Glu Glu Ile Thr Asp Glu Ser Val Ser Ser Ser Ser Glu 275 280 285 Ser Gly Ser Ser Thr Pro Gln Asp Gly Gly Ala Ala Ser Ser Gly Ala 290 295 300 Pro Ser Gly Asp Gln Ser Ile Ser Ala Asn Ala Cys Leu Ala Lys Ser 305 310 315 320 Tyr Ala Ala Ser Thr Asp Ser Ser Pro Val Ser Asn Ser Ser Gly Ser 325 330 335 Glu Glu Pro Val Thr Ser Ser Ser Asp Ser Asp Val Thr Ala Ser Ser 340 345 350 Asp Asn Pro Asp Ser Ser Ser Ser Gly Asp Ser Ala Gly Asp Ser Glu 355 360 365 Glu Pro Thr Glu Pro Glu Ala Gly Ser Thr Thr Glu Thr Leu Thr Leu 370 375 380 Ile Gly Gly Gly Ala Ile Tyr Gly Glu Thr Val Lys Ile Glu Asn Phe 385 390 395 400 Ser Gly Ala Ile Ser Met Arg Val Gly Tyr Tyr Gly Asp Phe Val Phe 405 410 415 Asp Arg Val Leu Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly Ala 420 425 430 Lys Pro Thr Thr Asp Thr Gly Asn Ser Ala Ala Pro Ser Thr Leu Thr 435 440 445 Ala Arg Glu Asn Pro Ala Tyr Gly Arg His Met Gln Asp Asn Met Phe 450 455 460 Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp 465 470 475 480 Thr Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr 485 490

495 Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Thr Gly 500 505 510 Ala Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu 515 520 525 Asn Ala Ile Ser Met Arg Met Gly Tyr Tyr Gly Asp Phe Val Phe Asp 530 535 540 Arg Val Leu Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly Asp Lys 545 550 555 560 Pro Thr Ser Thr Thr Gly Asn Ala Thr Ala Pro Thr Thr Leu Thr Ala 565 570 575 Arg Glu Asn Pro Ala Tyr Gly Arg His Met Gln Asp Asn Met Phe Thr 580 585 590 Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr 595 600 605 Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr 610 615 620 Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Ala Ser Ala 625 630 635 640 Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 645 650 655 Ala Ile Ser Met Arg Met Gly Tyr Tyr Gly Asp Phe Val Phe Asp Arg 660 665 670 Val Leu Lys Thr Asp Val Asn Lys Glu Phe Glu Met Gly Glu Ala Leu 675 680 685 Ala Gly Ala Ser Gly Asn Thr Thr Ser Thr Leu Ser Lys Leu Val Glu 690 695 700 Arg Thr Asn Pro Ala Tyr Gly Lys His Met Gln Asp Asn Met Phe Thr 705 710 715 720 Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ser Asp Thr 725 730 735 Ile Arg Ile Ala Gln Pro Arg Leu Val Thr Pro Val Val Asp Ile Thr 740 745 750 Thr Leu Asn Pro Thr Ile Ala Gly Ser Gly Ser Val Ala Gly Ala Asn 755 760 765 Thr Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln Leu 770 775 780 Asn 785 61740PRTChlamydia trachomatis 61Ser Arg Gln Asn Ala Glu Glu Asn Leu Lys Asn Phe Ala Lys Glu Leu 1 5 10 15 Lys Leu Pro Asp Val Ala Phe Asp Gln Asn Asn Thr Cys Ile Leu Phe 20 25 30 Val Asp Gly Glu Phe Ser Leu His Leu Thr Tyr Glu Glu His Ser Asp 35 40 45 Arg Leu Tyr Val Tyr Ala Pro Leu Leu Asp Gly Leu Pro Asp Asn Pro 50 55 60 Gln Arg Arg Leu Ala Leu Tyr Glu Lys Leu Leu Glu Gly Ser Met Leu 65 70 75 80 Gly Gly Gln Met Ala Gly Gly Gly Val Gly Val Ala Thr Lys Glu Gln 85 90 95 Leu Ile Leu Met His Cys Val Leu Asp Met Lys Tyr Ala Glu Thr Asn 100 105 110 Leu Leu Lys Ala Phe Ala Gln Leu Phe Ile Glu Thr Val Val Lys Trp 115 120 125 Arg Thr Val Cys Ser Asp Ile Ser Ala Gly Arg Glu Pro Thr Val Asp 130 135 140 Thr Met Pro Gln Met Pro Gln Gly Gly Gly Gly Gly Ile Gln Pro Pro 145 150 155 160 Pro Ala Gly Ile Arg Ala Thr Val Lys Ala Ile Val Glu Ser Thr Pro 165 170 175 Glu Ala Pro Glu Glu Ile Pro Pro Val Glu Gly Glu Glu Ser Thr Ala 180 185 190 Thr Glu Asp Pro Asn Ser Asn Thr Glu Gly Ser Ser Ala Asn Thr Asn 195 200 205 Leu Glu Gly Ser Gln Gly Asp Thr Ala Asp Thr Gly Thr Gly Asp Val 210 215 220 Asn Asn Glu Ser Gln Asp Thr Ser Asp Thr Gly Asn Ala Glu Ser Glu 225 230 235 240 Glu Gln Leu Gln Asp Ser Thr Gln Ser Asn Glu Glu Asn Thr Leu Pro 245 250 255 Asn Ser Asn Ile Asp Gln Ser Asn Glu Asn Thr Asp Glu Ser Ser Asp 260 265 270 Ser His Thr Glu Glu Ile Thr Asp Glu Ser Val Ser Ser Ser Ser Glu 275 280 285 Ser Gly Ser Ser Thr Pro Gln Asp Gly Gly Ala Ala Ser Ser Gly Ala 290 295 300 Pro Ser Gly Asp Gln Ser Ile Ser Ala Asn Ala Cys Leu Ala Lys Ser 305 310 315 320 Tyr Ala Ala Ser Thr Asp Ser Ser Pro Val Ser Asn Ser Ser Gly Ser 325 330 335 Glu Glu Pro Val Thr Ser Ser Ser Asp Ser Asp Val Thr Ala Ser Ser 340 345 350 Asp Asn Pro Asp Ser Ser Ser Ser Gly Asp Ser Ala Gly Asp Ser Glu 355 360 365 Glu Pro Thr Glu Pro Glu Ala Gly Ser Thr Thr Glu Thr Leu Thr Leu 370 375 380 Ile Gly Gly Gly Ala Ile Tyr Gly Glu Thr Val Lys Ile Glu Asn Phe 385 390 395 400 Ser Gly Asp Ala Ile Ser Met Arg Val Gly Tyr Tyr Gly Asp Phe Val 405 410 415 Phe Asp Arg Val Leu Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly 420 425 430 Ala Lys Pro Thr Thr Asp Thr Gly Asn Ser Ala Ala Pro Ser Thr Leu 435 440 445 Thr Ala Arg Glu Asn Pro Ala Tyr Gly Arg His Met Gln Asp Ala Glu 450 455 460 Met Phe Thr Asn Ala Ala Ser Met Ala Leu Asn Ile Trp Asp Arg Phe 465 470 475 480 Asp Val Phe Ser Thr Leu Gly Ala Thr Ser Gly Tyr Leu Lys Gly Asn 485 490 495 Ser Ala Ser Phe Asn Leu Val Gly Leu Phe Gly Asp Asn Glu Asn Gln 500 505 510 Lys Thr Val Lys Ala Glu Ser Val Pro Asn Met Ser Phe Asp Gln Ser 515 520 525 Val Val Glu Leu Tyr Thr Asp Thr Thr Phe Ala Trp Ser Val Gly Ala 530 535 540 Arg Ala Ala Leu Trp Glu Ser Gly Ser Ala Thr Leu Gly Ala Ser Phe 545 550 555 560 Gln Tyr Ala Gln Ser Lys Pro Lys Val Glu Glu Leu Asn Val Leu Ser 565 570 575 Asn Ala Ala Glu Phe Thr Ile Asn Lys Pro Lys Gly Tyr Val Gly Lys 580 585 590 Glu Phe Pro Leu Asp Leu Thr Ala Gly Thr Asp Ala Ala Thr Gly Thr 595 600 605 Lys Asp Ala Ser Ile Asp Tyr His Glu Trp Gln Ala Ser Leu Ala Leu 610 615 620 Ser Tyr Arg Leu Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser 625 630 635 640 Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser 645 650 655 Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly 660 665 670 Ala Gly Asp Val Lys Thr Gly Ala Glu Gly Gln Leu Gly Asp Thr Met 675 680 685 Gln Ile Val Ser Leu Gln Leu Asn Lys Met Lys Ser Arg Lys Ser Ser 690 695 700 Gly Ile Ala Val Gly Thr Thr Ile Val Asp Ala Asp Lys Tyr Ala Val 705 710 715 720 Thr Val Glu Thr Arg Leu Ile Asp Glu Arg Ala Ala His Val Asn Ala 725 730 735 Gln Phe Arg Phe 740 62294PRTChlamydia trachomatis 62Asp Ala Ile Ser Met Arg Val Gly Tyr Tyr Gly Asp Phe Val Phe Asp 1 5 10 15 Arg Val Leu Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly Ala Lys 20 25 30 Pro Thr Thr Asp Thr Gly Asn Ser Ala Ala Pro Ser Thr Leu Thr Ala 35 40 45 Arg Glu Asn Pro Ala Tyr Gly Arg His Met Gln Asp Ala Glu Met Phe 50 55 60 Thr Asn Ala Ala Cys Met Ala Leu Asn Ile Trp Asp Arg Phe Asp Val 65 70 75 80 Phe Cys Thr Leu Gly Ala Thr Ser Gly Tyr Leu Lys Gly Asn Ser Ala 85 90 95 Ser Phe Asn Leu Val Gly Leu Phe Gly Asp Asn Glu Asn Gln Lys Thr 100 105 110 Val Lys Ala Glu Ser Val Pro Asn Met Ser Phe Asp Gln Ser Val Val 115 120 125 Glu Leu Tyr Thr Asp Thr Thr Phe Ala Trp Ser Val Gly Ala Arg Ala 130 135 140 Ala Leu Trp Glu Cys Gly Cys Ala Thr Leu Gly Ala Ser Phe Gln Tyr 145 150 155 160 Ala Gln Ser Lys Pro Lys Val Glu Glu Leu Asn Val Leu Cys Asn Ala 165 170 175 Ala Glu Phe Thr Ile Asn Lys Pro Lys Gly Tyr Val Gly Lys Glu Phe 180 185 190 Pro Leu Asp Leu Thr Ala Gly Thr Asp Ala Ala Thr Gly Thr Lys Asp 195 200 205 Ala Ser Ile Asp Tyr His Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr 210 215 220 Arg Leu Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala 225 230 235 240 Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr 245 250 255 Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly 260 265 270 Asp Val Lys Thr Gly Ala Glu Gly Gln Leu Gly Asp Thr Met Gln Ile 275 280 285 Val Ser Leu Gln Leu Asn 290 63294PRTChlamydia trachomatis 63Asp Ala Ile Ser Met Arg Val Gly Tyr Tyr Gly Asp Phe Val Phe Asp 1 5 10 15 Arg Val Leu Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly Ala Lys 20 25 30 Pro Thr Thr Asp Thr Gly Asn Ser Ala Ala Pro Ser Thr Leu Thr Ala 35 40 45 Arg Glu Asn Pro Ala Tyr Gly Arg His Met Gln Asp Ala Glu Met Phe 50 55 60 Thr Asn Ala Ala Ser Met Ala Leu Asn Ile Trp Asp Arg Phe Asp Val 65 70 75 80 Phe Ser Thr Leu Gly Ala Thr Ser Gly Tyr Leu Lys Gly Asn Ser Ala 85 90 95 Ser Phe Asn Leu Val Gly Leu Phe Gly Asp Asn Glu Asn Gln Lys Thr 100 105 110 Val Lys Ala Glu Ser Val Pro Asn Met Ser Phe Asp Gln Ser Val Val 115 120 125 Glu Leu Tyr Thr Asp Thr Thr Phe Ala Trp Ser Val Gly Ala Arg Ala 130 135 140 Ala Leu Trp Glu Ser Gly Ser Ala Thr Leu Gly Ala Ser Phe Gln Tyr 145 150 155 160 Ala Gln Ser Lys Pro Lys Val Glu Glu Leu Asn Val Leu Ser Asn Ala 165 170 175 Ala Glu Phe Thr Ile Asn Lys Pro Lys Gly Tyr Val Gly Lys Glu Phe 180 185 190 Pro Leu Asp Leu Thr Ala Gly Thr Asp Ala Ala Thr Gly Thr Lys Asp 195 200 205 Ala Ser Ile Asp Tyr His Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr 210 215 220 Arg Leu Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala 225 230 235 240 Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr 245 250 255 Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly 260 265 270 Asp Val Lys Thr Gly Ala Glu Gly Gln Leu Gly Asp Thr Met Gln Ile 275 280 285 Val Ser Leu Gln Leu Asn 290 64500PRTChlamydia trachomatis 64Asp Ala Ile Ser Met Arg Val Gly Tyr Tyr Gly Asp Phe Val Phe Asp 1 5 10 15 Arg Val Leu Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly Ala Lys 20 25 30 Pro Thr Thr Asp Thr Gly Asn Ser Ala Ala Pro Ser Thr Leu Thr Ala 35 40 45 Arg Glu Asn Pro Ala Tyr Gly Arg His Met Gln Asp Ala Glu Met Phe 50 55 60 Thr Asn Ala Ala Ser Met Ala Leu Asn Ile Trp Asp Arg Phe Asp Val 65 70 75 80 Phe Ser Thr Leu Gly Ala Thr Ser Gly Tyr Leu Lys Gly Asn Ser Ala 85 90 95 Ser Phe Asn Leu Val Gly Leu Phe Gly Asp Asn Glu Asn Gln Lys Thr 100 105 110 Val Lys Ala Glu Ser Val Pro Asn Met Ser Phe Asp Gln Ser Val Val 115 120 125 Glu Leu Tyr Thr Asp Thr Thr Phe Ala Trp Ser Val Gly Ala Arg Ala 130 135 140 Ala Leu Trp Glu Ser Gly Ser Ala Thr Leu Gly Ala Ser Phe Gln Tyr 145 150 155 160 Ala Gln Ser Lys Pro Lys Val Glu Glu Leu Asn Val Leu Ser Asn Ala 165 170 175 Ala Glu Phe Thr Ile Asn Lys Pro Lys Gly Tyr Val Gly Lys Glu Phe 180 185 190 Pro Leu Asp Leu Thr Ala Gly Thr Asp Ala Ala Thr Gly Thr Lys Asp 195 200 205 Ala Ser Ile Asp Tyr His Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr 210 215 220 Arg Leu Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala 225 230 235 240 Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr 245 250 255 Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly 260 265 270 Asp Val Lys Thr Gly Ala Glu Gly Gln Leu Gly Asp Thr Met Gln Ile 275 280 285 Val Ser Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr Ile Gly Val Lys 290 295 300 Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro 305 310 315 320 Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile 325 330 335 Ala Gly Ala Gly Asp Val Lys Ala Ser Ala Glu Gly Gln Leu Gly Asp 340 345 350 Thr Met Gln Ile Val Ser Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr 355 360 365 Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ser Asp Thr Ile Arg 370 375 380 Ile Ala Gln Pro Arg Leu Val Thr Pro Val Val Asp Ile Thr Thr Leu 385 390 395 400 Asn Pro Thr Ile Ala Gly Ser Gly Ser Val Ala Gly Ala Asn Thr Glu 405 410 415 Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Asn 420 425 430 Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp 435 440 445 Ser Asn Thr Ile Arg Ile Ala Gln Pro Lys Leu Ala Lys Pro Val Val 450 455 460 Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala Gly Ser Gly Ser Val Val 465 470 475 480 Ala Ala Asn Ser Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser 485 490 495 Leu Gln Leu Asn 500 651068PRTChlamydia trachomatis 65Thr Val Lys Ala Ile Val Glu Ser Thr Pro Glu Ala Pro Glu Glu Ile 1 5 10 15 Pro Pro Val Glu Gly Glu Glu Ser Thr Ala Thr Glu Asp Pro Asn Ser 20 25 30 Asn Thr Glu Gly Ser Ser Ala Asn Thr Asn Leu Glu Gly Ser Gln Gly 35 40 45 Asp Thr Ala Asp Thr Gly Thr Gly Asp Val Asn Asn Glu Ser Gln Asp 50 55 60 Thr Ser Asp Thr Gly Asn Ala Glu Ser Glu Glu Gln Leu Gln Asp Ser 65 70 75 80 Thr Gln Ser Asn Glu Glu Asn Thr Leu Pro Asn Ser Asn Ile Asp Gln 85 90 95 Ser Asn Glu Asn Thr Asp Glu Ser Ser Asp Ser His Thr Glu Glu Ile 100 105 110 Thr Asp Glu Ser Val Ser Ser Ser Ser Glu Ser Gly Ser Ser Thr Pro 115 120 125 Gln Asp Gly Gly Ala Ala Ser Ser Gly Ala Pro Ser Gly Asp Gln Ser 130 135 140 Ile Ser Ala Asn Ala Cys Leu Ala Lys Ser Tyr Ala Ala Ser Thr Asp 145 150 155 160 Ser Ser Pro

Val Ser Asn Ser Ser Gly Ser Glu Glu Pro Val Thr Ser 165 170 175 Ser Ser Asp Ser Asp Val Thr Ala Ser Ser Asp Asn Pro Asp Ser Ser 180 185 190 Ser Ser Gly Asp Ser Ala Gly Asp Ser Glu Glu Pro Thr Glu Pro Glu 195 200 205 Ala Gly Ser Thr Thr Glu Thr Leu Thr Leu Ile Gly Gly Gly Ala Ile 210 215 220 Tyr Gly Glu Thr Val Lys Ile Glu Asn Phe Ser Gly Ser Arg Gln Asn 225 230 235 240 Ala Glu Glu Asn Leu Lys Asn Phe Ala Lys Glu Leu Lys Leu Pro Asp 245 250 255 Val Ala Phe Asp Gln Asn Asn Thr Cys Ile Leu Phe Val Asp Gly Glu 260 265 270 Phe Ser Leu His Leu Thr Tyr Glu Glu His Ser Asp Arg Leu Tyr Val 275 280 285 Tyr Ala Pro Leu Leu Asp Gly Leu Pro Asp Asn Pro Gln Arg Arg Leu 290 295 300 Ala Leu Tyr Glu Lys Leu Leu Glu Gly Ser Met Leu Gly Gly Gln Met 305 310 315 320 Ala Gly Gly Gly Val Gly Val Ala Thr Lys Glu Gln Leu Ile Leu Met 325 330 335 His Cys Val Leu Asp Met Lys Tyr Ala Glu Thr Asn Leu Leu Lys Ala 340 345 350 Phe Ala Gln Leu Phe Ile Glu Thr Val Val Lys Trp Arg Thr Val Cys 355 360 365 Ser Asp Ile Ser Ala Gly Arg Glu Pro Thr Val Asp Thr Met Pro Gln 370 375 380 Met Pro Gln Gly Gly Gly Gly Gly Ile Gln Pro Pro Pro Ala Gly Ile 385 390 395 400 Arg Ala Ser Arg Gln Asn Ala Glu Glu Asn Leu Lys Asn Phe Ala Lys 405 410 415 Glu Leu Lys Leu Pro Asp Val Ala Phe Asp Gln Asn Asn Thr Cys Ile 420 425 430 Leu Phe Val Asp Gly Glu Phe Ser Leu His Leu Thr Tyr Glu Glu His 435 440 445 Ser Asp Arg Leu Tyr Val Tyr Ala Pro Leu Leu Asp Gly Leu Pro Asp 450 455 460 Asn Pro Gln Arg Arg Leu Ala Leu Tyr Glu Lys Leu Leu Glu Gly Ser 465 470 475 480 Met Leu Gly Gly Gln Met Ala Gly Gly Gly Val Gly Val Ala Thr Lys 485 490 495 Glu Gln Leu Ile Leu Met His Cys Val Leu Asp Met Lys Tyr Ala Glu 500 505 510 Thr Asn Leu Leu Lys Ala Phe Ala Gln Leu Phe Ile Glu Thr Val Val 515 520 525 Lys Trp Arg Thr Val Cys Ser Asp Ile Ser Ala Gly Arg Glu Pro Thr 530 535 540 Val Asp Thr Met Pro Gln Met Pro Gln Gly Gly Gly Gly Gly Ile Gln 545 550 555 560 Pro Pro Pro Ala Gly Ile Arg Ala Asp Ala Ile Ser Met Arg Val Gly 565 570 575 Tyr Tyr Gly Asp Phe Val Phe Asp Arg Val Leu Lys Thr Asp Val Asn 580 585 590 Lys Glu Phe Gln Met Gly Ala Lys Pro Thr Thr Asp Thr Gly Asn Ser 595 600 605 Ala Ala Pro Ser Thr Leu Thr Ala Arg Glu Asn Pro Ala Tyr Gly Arg 610 615 620 His Met Gln Asp Ala Glu Met Phe Thr Asn Ala Ala Ser Met Ala Leu 625 630 635 640 Asn Ile Trp Asp Arg Phe Asp Val Phe Ser Thr Leu Gly Ala Thr Ser 645 650 655 Gly Tyr Leu Lys Gly Asn Ser Ala Ser Phe Asn Leu Val Gly Leu Phe 660 665 670 Gly Asp Asn Glu Asn Gln Lys Thr Val Lys Ala Glu Ser Val Pro Asn 675 680 685 Met Ser Phe Asp Gln Ser Val Val Glu Leu Tyr Thr Asp Thr Thr Phe 690 695 700 Ala Trp Ser Val Gly Ala Arg Ala Ala Leu Trp Glu Ser Gly Ser Ala 705 710 715 720 Thr Leu Gly Ala Ser Phe Gln Tyr Ala Gln Ser Lys Pro Lys Val Glu 725 730 735 Glu Leu Asn Val Leu Ser Asn Ala Ala Glu Phe Thr Ile Asn Lys Pro 740 745 750 Lys Gly Tyr Val Gly Lys Glu Phe Pro Leu Asp Leu Thr Ala Gly Thr 755 760 765 Asp Ala Ala Thr Gly Thr Lys Asp Ala Ser Ile Asp Tyr His Glu Trp 770 775 780 Gln Ala Ser Leu Ala Leu Ser Tyr Arg Leu Asn Met Phe Thr Pro Tyr 785 790 795 800 Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg 805 810 815 Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu 820 825 830 Asn Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Thr Gly Ala Glu Gly 835 840 845 Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Asn Met 850 855 860 Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala 865 870 875 880 Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp 885 890 895 Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Ala 900 905 910 Ser Ala Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln 915 920 925 Leu Asn Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala 930 935 940 Ser Phe Asp Ser Asp Thr Ile Arg Ile Ala Gln Pro Arg Leu Val Thr 945 950 955 960 Pro Val Val Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala Gly Ser Gly 965 970 975 Ser Val Ala Gly Ala Asn Thr Glu Gly Gln Ile Ser Asp Thr Met Gln 980 985 990 Ile Val Ser Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr Ile Gly Val 995 1000 1005 Lys Trp Ser Arg Ala Ser Phe Asp Ser Asn Thr Ile Arg Ile Ala 1010 1015 1020 Gln Pro Lys Leu Ala Lys Pro Val Val Asp Ile Thr Thr Leu Asn 1025 1030 1035 Pro Thr Ile Ala Gly Ser Gly Ser Val Val Ala Ala Asn Ser Glu 1040 1045 1050 Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 1055 1060 1065 66606PRTChlamydia trachomatis 66Ser Arg Gln Asn Ala Glu Glu Asn Leu Lys Asn Phe Ala Lys Glu Leu 1 5 10 15 Lys Leu Pro Asp Val Ala Phe Asp Gln Asn Asn Thr Cys Ile Leu Phe 20 25 30 Val Asp Gly Glu Phe Ser Leu His Leu Thr Tyr Glu Glu His Ser Asp 35 40 45 Arg Leu Tyr Val Tyr Ala Pro Leu Leu Asp Gly Leu Pro Asp Asn Pro 50 55 60 Gln Arg Arg Leu Ala Leu Tyr Glu Lys Leu Leu Glu Gly Ser Met Leu 65 70 75 80 Gly Gly Gln Met Ala Gly Gly Gly Val Gly Val Ala Thr Lys Glu Gln 85 90 95 Leu Ile Leu Met His Cys Val Leu Asp Met Lys Tyr Ala Glu Thr Asn 100 105 110 Leu Leu Lys Ala Phe Ala Gln Leu Phe Ile Glu Thr Val Val Lys Trp 115 120 125 Arg Thr Val Cys Ser Asp Ile Ser Ala Gly Arg Glu Pro Thr Val Asp 130 135 140 Thr Met Pro Gln Met Pro Gln Gly Gly Gly Gly Gly Ile Gln Pro Pro 145 150 155 160 Pro Ala Gly Ile Arg Ala Ser Arg Gln Asn Ala Glu Glu Asn Leu Lys 165 170 175 Asn Phe Ala Lys Glu Leu Lys Leu Pro Asp Val Ala Phe Asp Gln Asn 180 185 190 Asn Thr Cys Ile Leu Phe Val Asp Gly Glu Phe Ser Leu His Leu Thr 195 200 205 Tyr Glu Glu His Ser Asp Arg Leu Tyr Val Tyr Ala Pro Leu Leu Asp 210 215 220 Gly Leu Pro Asp Asn Pro Gln Arg Arg Leu Ala Leu Tyr Glu Lys Leu 225 230 235 240 Leu Glu Gly Ser Met Leu Gly Gly Gln Met Ala Gly Gly Gly Val Gly 245 250 255 Val Ala Thr Lys Glu Gln Leu Ile Leu Met His Cys Val Leu Asp Met 260 265 270 Lys Tyr Ala Glu Thr Asn Leu Leu Lys Ala Phe Ala Gln Leu Phe Ile 275 280 285 Glu Thr Val Val Lys Trp Arg Thr Val Cys Ser Asp Ile Ser Ala Gly 290 295 300 Arg Glu Pro Thr Val Asp Thr Met Pro Gln Met Pro Gln Gly Gly Gly 305 310 315 320 Gly Gly Ile Gln Pro Pro Pro Ala Gly Ile Arg Ala Asn Met Phe Thr 325 330 335 Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr 340 345 350 Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr 355 360 365 Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Thr Gly Ala 370 375 380 Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 385 390 395 400 Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe 405 410 415 Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile 420 425 430 Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val 435 440 445 Lys Ala Ser Ala Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser 450 455 460 Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser 465 470 475 480 Arg Ala Ser Phe Asp Ser Asp Thr Ile Arg Ile Ala Gln Pro Arg Leu 485 490 495 Val Thr Pro Val Val Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala Gly 500 505 510 Ser Gly Ser Val Ala Gly Ala Asn Thr Glu Gly Gln Ile Ser Asp Thr 515 520 525 Met Gln Ile Val Ser Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr Ile 530 535 540 Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ser Asn Thr Ile Arg Ile 545 550 555 560 Ala Gln Pro Lys Leu Ala Lys Pro Val Val Asp Ile Thr Thr Leu Asn 565 570 575 Pro Thr Ile Ala Gly Ser Gly Ser Val Val Ala Ala Asn Ser Glu Gly 580 585 590 Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 595 600 605 671093PRTChlamydia trachomatis 67Ser Arg Gln Asn Ala Glu Glu Asn Leu Lys Asn Phe Ala Lys Glu Leu 1 5 10 15 Lys Leu Pro Asp Val Ala Phe Asp Gln Asn Asn Thr Cys Ile Leu Phe 20 25 30 Val Asp Gly Glu Phe Ser Leu His Leu Thr Tyr Glu Glu His Ser Asp 35 40 45 Arg Leu Tyr Val Tyr Ala Pro Leu Leu Asp Gly Leu Pro Asp Asn Pro 50 55 60 Gln Arg Arg Leu Ala Leu Tyr Glu Lys Leu Leu Glu Gly Ser Met Leu 65 70 75 80 Gly Gly Gln Met Ala Gly Gly Gly Val Gly Val Ala Thr Lys Glu Gln 85 90 95 Leu Ile Leu Met His Cys Val Leu Asp Met Lys Tyr Ala Glu Thr Asn 100 105 110 Leu Leu Lys Ala Phe Ala Gln Leu Phe Ile Glu Thr Val Val Lys Trp 115 120 125 Arg Thr Val Cys Ser Asp Ile Ser Ala Gly Arg Glu Pro Thr Val Asp 130 135 140 Thr Met Pro Gln Met Pro Gln Gly Gly Gly Gly Gly Ile Gln Pro Pro 145 150 155 160 Pro Ala Gly Ile Arg Ala Ser Arg Gln Asn Ala Glu Glu Asn Leu Lys 165 170 175 Asn Phe Ala Lys Glu Leu Lys Leu Pro Asp Val Ala Phe Asp Gln Asn 180 185 190 Asn Thr Cys Ile Leu Phe Val Asp Gly Glu Phe Ser Leu His Leu Thr 195 200 205 Tyr Glu Glu His Ser Asp Arg Leu Tyr Val Tyr Ala Pro Leu Leu Asp 210 215 220 Gly Leu Pro Asp Asn Pro Gln Arg Arg Leu Ala Leu Tyr Glu Lys Leu 225 230 235 240 Leu Glu Gly Ser Met Leu Gly Gly Gln Met Ala Gly Gly Gly Val Gly 245 250 255 Val Ala Thr Lys Glu Gln Leu Ile Leu Met His Cys Val Leu Asp Met 260 265 270 Lys Tyr Ala Glu Thr Asn Leu Leu Lys Ala Phe Ala Gln Leu Phe Ile 275 280 285 Glu Thr Val Val Lys Trp Arg Thr Val Cys Ser Asp Ile Ser Ala Gly 290 295 300 Arg Glu Pro Thr Val Asp Thr Met Pro Gln Met Pro Gln Gly Gly Gly 305 310 315 320 Gly Gly Ile Gln Pro Pro Pro Ala Gly Ile Arg Ala Gly Ile Ala His 325 330 335 Thr Glu Trp Glu Ser Val Ile Gly Leu Glu Val His Val Glu Leu Asn 340 345 350 Thr Glu Ser Lys Leu Phe Ser Pro Ala Arg Asn His Phe Gly Asp Glu 355 360 365 Pro Asn Thr Asn Ile Ser Pro Val Cys Thr Gly Met Pro Gly Ser Leu 370 375 380 Pro Val Leu Asn Lys Asp Ala Val Arg Lys Ala Val Leu Phe Gly Cys 385 390 395 400 Ala Val Glu Gly Asp Val Ala Leu Phe Ser Arg Phe Asp Arg Lys Ser 405 410 415 Tyr Phe Tyr Pro Asp Ser Pro Arg Asn Phe Gln Ile Thr Gln Tyr Glu 420 425 430 His Pro Ile Val Arg Gly Gly Cys Ile Arg Ala Val Val Glu Gly Glu 435 440 445 Glu Lys Thr Phe Glu Leu Ala Gln Thr His Leu Glu Asp Asp Ala Gly 450 455 460 Met Leu Lys His Phe Gly Asp Phe Ala Gly Val Asp Tyr Asn Arg Ala 465 470 475 480 Gly Val Pro Leu Ile Glu Ile Val Ser Lys Pro Cys Met Phe Ser Ala 485 490 495 Glu Asp Ala Val Ala Tyr Ala Asn Ala Leu Val Ser Ile Leu Gly Tyr 500 505 510 Ile Gly Ile Ser Asp Cys Asn Met Glu Glu Gly Ser Ile Arg Phe Asp 515 520 525 Val Asn Ile Ser Val Arg Pro Arg Gly Ser Arg Glu Leu Arg Asn Lys 530 535 540 Val Glu Ile Lys Asn Met Asn Ser Phe Thr Phe Met Ala Gln Ala Leu 545 550 555 560 Glu Ala Glu Lys Arg Arg Gln Ile Glu Glu Tyr Leu Ser Tyr Pro Asn 565 570 575 Glu Asp Pro Lys Lys Val Val Pro Ala Ala Thr Tyr Arg Trp Asp Pro 580 585 590 Glu Lys Lys Lys Thr Val Leu Met Arg Leu Lys Glu Arg Ala Glu Asp 595 600 605 Tyr Met Tyr Phe Val Glu Pro Asp Leu Pro Val Leu Gln Ile Thr Glu 610 615 620 Thr Tyr Ile Asp Glu Val Arg Gln Thr Leu Pro Glu Leu Pro His Ser 625 630 635 640 Lys Tyr Met Arg Tyr Ile Thr Asp Phe Asp Ile Ala Glu Asp Leu Ala 645 650 655 Met Ile Leu Val Gly Asp Arg His Thr Ala His Phe Phe Glu Thr Ala 660 665 670 Thr Met Ser Cys Lys Asn Tyr Arg Ala Leu Ser Asn Trp Ile Thr Val 675 680 685 Glu Phe Ala Gly Arg Cys Lys Ala Arg Gly Lys Thr Leu Pro Phe Thr 690 695 700 Gly Ile Leu Pro Glu Trp Val Ala Gln Leu Val Asn Phe Ile Asp Arg 705 710 715 720 Gly Val Ile Thr Gly Lys Ile Ala Lys Glu Ile Ala Asp Arg Met Val 725 730 735 Ser Ser Phe Gly Glu Ser Pro Glu Asp Ile Leu Arg Arg His Pro Ser 740 745 750 Leu Leu Pro Met Thr Asp Asp His Ala Leu Arg Ala Ile Val Lys Glu 755 760 765 Val Val Ala Gln Asn Thr Ala Ser Val Ala Asp Tyr Lys Asn Gly Lys 770 775

780 Ala Lys Ala Leu Gly Phe Leu Val Gly Gln Ile Met Lys Arg Thr Glu 785 790 795 800 Gly Lys Ala Pro Pro Lys Arg Val Asn Glu Leu Leu Leu Ala Ala Met 805 810 815 Arg Asp Met Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg 820 825 830 Ala Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys Ser Ala 835 840 845 Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala 850 855 860 Gly Asp Val Lys Thr Gly Ala Glu Gly Gln Leu Gly Asp Thr Met Gln 865 870 875 880 Ile Val Ser Leu Gln Leu Asn Asn Met Phe Thr Pro Tyr Ile Gly Val 885 890 895 Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln 900 905 910 Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr 915 920 925 Ile Ala Gly Ala Gly Asp Val Lys Ala Ser Ala Glu Gly Gln Leu Gly 930 935 940 Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Asn Met Phe Thr Pro 945 950 955 960 Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ser Asp Thr Ile 965 970 975 Arg Ile Ala Gln Pro Arg Leu Val Thr Pro Val Val Asp Ile Thr Thr 980 985 990 Leu Asn Pro Thr Ile Ala Gly Ser Gly Ser Val Ala Gly Ala Asn Thr 995 1000 1005 Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln Leu 1010 1015 1020 Asn Asn Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala 1025 1030 1035 Ser Phe Asp Ser Asn Thr Ile Arg Ile Ala Gln Pro Lys Leu Ala 1040 1045 1050 Lys Pro Val Val Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala Gly 1055 1060 1065 Ser Gly Ser Val Val Ala Ala Asn Ser Glu Gly Gln Ile Ser Asp 1070 1075 1080 Thr Met Gln Ile Val Ser Leu Gln Leu Asn 1085 1090 68393PRTChlamydia trachomatis 68Met Lys Lys Leu Leu Lys Ser Val Leu Val Phe Ala Ala Leu Ser Ser 1 5 10 15 Ala Ser Ser Leu Gln Ala Leu Pro Val Gly Asn Pro Ala Glu Pro Ser 20 25 30 Leu Met Ile Asp Gly Ile Leu Trp Glu Gly Phe Gly Gly Asp Pro Cys 35 40 45 Asp Pro Cys Ala Thr Trp Cys Asp Ala Ile Ser Met Arg Val Gly Tyr 50 55 60 Tyr Gly Asp Phe Val Phe Asp Arg Val Leu Lys Thr Asp Val Asn Lys 65 70 75 80 Glu Phe Gln Met Gly Ala Lys Pro Thr Thr Asp Thr Gly Asn Ser Ala 85 90 95 Ala Pro Ser Thr Leu Thr Ala Arg Glu Asn Pro Ala Tyr Gly Arg His 100 105 110 Met Gln Asp Ala Glu Met Phe Thr Asn Ala Ala Cys Met Ala Leu Asn 115 120 125 Ile Trp Asp Arg Phe Asp Val Phe Cys Thr Leu Gly Ala Thr Ser Gly 130 135 140 Tyr Leu Lys Gly Asn Ser Ala Ser Phe Asn Leu Val Gly Leu Phe Gly 145 150 155 160 Asp Asn Glu Asn Gln Lys Thr Val Lys Ala Glu Ser Val Pro Asn Met 165 170 175 Ser Phe Asp Gln Ser Val Val Glu Leu Tyr Thr Asp Thr Thr Phe Ala 180 185 190 Trp Ser Val Gly Ala Arg Ala Ala Leu Trp Glu Cys Gly Cys Ala Thr 195 200 205 Leu Gly Ala Ser Phe Gln Tyr Ala Gln Ser Lys Pro Lys Val Glu Glu 210 215 220 Leu Asn Val Leu Cys Asn Ala Ala Glu Phe Thr Ile Asn Lys Pro Lys 225 230 235 240 Gly Tyr Val Gly Lys Glu Phe Pro Leu Asp Leu Thr Ala Gly Thr Asp 245 250 255 Ala Ala Thr Gly Thr Lys Asp Ala Ser Ile Asp Tyr His Glu Trp Gln 260 265 270 Ala Ser Leu Ala Leu Ser Tyr Arg Leu Asn Met Phe Thr Pro Tyr Ile 275 280 285 Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg Ile 290 295 300 Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn 305 310 315 320 Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Thr Gly Ala Glu Gly Gln 325 330 335 Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Lys Met Lys 340 345 350 Ser Arg Lys Ser Cys Gly Ile Ala Val Gly Thr Thr Ile Val Asp Ala 355 360 365 Asp Lys Tyr Ala Val Thr Val Glu Thr Arg Leu Ile Asp Glu Arg Ala 370 375 380 Ala His Val Asn Ala Gln Phe Arg Phe 385 390 69122PRTChlamydia trachomatis 69Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala 1 5 10 15 Gly Ala Gly Asp Val Lys Thr Gly Ala Glu Gly Gln Leu Gly Ser Ala 20 25 30 Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala 35 40 45 Gly Asp Val Lys Ala Ser Ala Glu Gly Gln Leu Gly Leu Val Thr Pro 50 55 60 Val Val Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala Gly Ser Gly Ser 65 70 75 80 Val Ala Gly Ala Asn Thr Glu Gly Gln Ile Ser Leu Ala Lys Pro Val 85 90 95 Val Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala Gly Ser Gly Ser Val 100 105 110 Val Ala Ala Asn Ser Glu Gly Gln Ile Ser 115 120 70178PRTChlamydia trachomatis 70Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr 1 5 10 15 Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Thr Gly Ala 20 25 30 Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser Ile Arg Ile Ala 35 40 45 Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn Pro 50 55 60 Thr Ile Ala Gly Ala Gly Asp Val Lys Ala Ser Ala Glu Gly Gln Leu 65 70 75 80 Gly Asp Thr Met Gln Ile Val Ser Ile Arg Ile Ala Gln Pro Arg Leu 85 90 95 Val Thr Pro Val Val Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala Gly 100 105 110 Ser Gly Ser Val Ala Gly Ala Asn Thr Glu Gly Gln Ile Ser Asp Thr 115 120 125 Met Gln Ile Val Ser Ile Arg Ile Ala Gln Pro Lys Leu Ala Lys Pro 130 135 140 Val Val Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala Gly Ser Gly Ser 145 150 155 160 Val Val Ala Ala Asn Ser Glu Gly Gln Ile Ser Asp Thr Met Gln Ile 165 170 175 Val Ser 71393PRTChlamydia trachomatis 71Met Lys Lys Leu Leu Lys Ser Val Leu Val Phe Ala Ala Leu Ser Ser 1 5 10 15 Ala Ser Ser Leu Gln Ala Leu Pro Val Gly Asn Pro Ala Glu Pro Ser 20 25 30 Leu Met Ile Asp Gly Ile Leu Trp Glu Gly Phe Gly Gly Asp Pro Cys 35 40 45 Asp Pro Cys Thr Thr Trp Cys Asp Ala Ile Ser Met Arg Met Gly Tyr 50 55 60 Tyr Gly Asp Phe Val Phe Asp Arg Val Leu Lys Thr Asp Val Asn Lys 65 70 75 80 Glu Phe Gln Met Gly Asp Lys Pro Thr Ser Thr Thr Gly Asn Ala Thr 85 90 95 Ala Pro Thr Thr Leu Thr Ala Arg Glu Asn Pro Ala Tyr Gly Arg His 100 105 110 Met Gln Asp Ala Glu Met Phe Thr Asn Ala Ala Cys Met Ala Leu Asn 115 120 125 Ile Trp Asp Arg Phe Asp Val Phe Cys Thr Leu Gly Ala Ser Ser Gly 130 135 140 Tyr Leu Lys Gly Asn Ser Ala Ser Phe Asn Leu Val Gly Leu Phe Gly 145 150 155 160 Asp Asn Glu Asn Gln Ser Thr Val Lys Thr Asn Ser Val Pro Asn Met 165 170 175 Ser Leu Asp Gln Ser Val Val Glu Leu Tyr Thr Asp Thr Ala Phe Ser 180 185 190 Trp Ser Val Gly Ala Arg Ala Ala Leu Trp Glu Cys Gly Cys Ala Thr 195 200 205 Leu Gly Ala Ser Phe Gln Tyr Ala Gln Ser Lys Pro Lys Val Glu Glu 210 215 220 Leu Asn Val Leu Cys Asn Ala Ala Glu Phe Thr Ile Asn Lys Pro Lys 225 230 235 240 Gly Tyr Val Gly Gln Glu Phe Pro Leu Ala Leu Ile Ala Gly Thr Asp 245 250 255 Ala Ala Thr Gly Thr Lys Asp Ala Ser Ile Asp Tyr His Glu Trp Gln 260 265 270 Ala Ser Leu Ala Leu Ser Tyr Arg Leu Asn Met Phe Thr Pro Tyr Ile 275 280 285 Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg Ile 290 295 300 Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr Leu Asn 305 310 315 320 Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Ala Ser Ala Glu Gly Gln 325 330 335 Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Lys Met Lys 340 345 350 Ser Arg Lys Ser Cys Gly Ile Ala Val Gly Thr Thr Ile Val Asp Ala 355 360 365 Asp Lys Tyr Ala Val Thr Val Glu Thr Arg Leu Ile Asp Glu Arg Ala 370 375 380 Ala His Val Asn Ala Gln Phe Arg Phe 385 390 72395PRTChlamydia trachomatis 72Met Lys Lys Leu Leu Lys Ser Val Leu Val Phe Ala Ala Leu Ser Ser 1 5 10 15 Ala Ser Ser Leu Gln Ala Leu Pro Val Gly Asn Pro Ala Glu Pro Ser 20 25 30 Leu Met Ile Asp Gly Ile Leu Trp Glu Gly Phe Gly Gly Asp Pro Cys 35 40 45 Asp Pro Cys Thr Thr Trp Cys Asp Ala Ile Ser Met Arg Met Gly Tyr 50 55 60 Tyr Gly Asp Phe Val Phe Asp Arg Val Leu Lys Thr Asp Val Asn Lys 65 70 75 80 Glu Phe Glu Met Gly Glu Ala Leu Ala Gly Ala Ser Gly Asn Thr Thr 85 90 95 Ser Thr Leu Ser Lys Leu Val Glu Arg Thr Asn Pro Ala Tyr Gly Lys 100 105 110 His Met Gln Asp Ala Glu Met Phe Thr Asn Ala Ala Cys Met Thr Leu 115 120 125 Asn Ile Trp Asp Arg Phe Asp Val Phe Cys Thr Leu Gly Ala Thr Ser 130 135 140 Gly Tyr Leu Lys Gly Asn Ser Ala Ser Phe Asn Leu Val Gly Leu Phe 145 150 155 160 Gly Asp Gly Val Asn Ala Thr Lys Pro Ala Ala Asp Ser Ile Pro Asn 165 170 175 Val Gln Leu Asn Gln Ser Val Val Glu Leu Tyr Thr Asp Thr Thr Phe 180 185 190 Ala Trp Ser Val Gly Ala Arg Ala Ala Leu Trp Glu Cys Gly Cys Ala 195 200 205 Thr Leu Gly Ala Ser Phe Gln Tyr Ala Gln Ser Lys Pro Lys Ile Glu 210 215 220 Glu Leu Asn Val Leu Cys Asn Ala Ala Glu Phe Thr Ile Asn Lys Pro 225 230 235 240 Lys Gly Tyr Val Gly Lys Glu Phe Pro Leu Asp Leu Thr Ala Gly Thr 245 250 255 Asp Ala Ala Thr Gly Thr Lys Asp Ala Ser Ile Asp Tyr His Glu Trp 260 265 270 Gln Ala Ser Leu Ser Leu Ser Tyr Arg Leu Asn Met Phe Thr Pro Tyr 275 280 285 Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ser Asp Thr Ile Arg 290 295 300 Ile Ala Gln Pro Arg Leu Val Thr Pro Val Val Asp Ile Thr Thr Leu 305 310 315 320 Asn Pro Thr Ile Ala Gly Cys Gly Ser Val Ala Gly Ala Asn Thr Glu 325 330 335 Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Lys 340 345 350 Met Lys Ser Arg Lys Ser Cys Gly Ile Ala Val Gly Thr Thr Ile Val 355 360 365 Asp Ala Asp Lys Tyr Ala Val Thr Val Glu Thr Arg Leu Ile Asp Glu 370 375 380 Arg Ala Ala His Val Asn Ala Gln Phe Arg Phe 385 390 395 73393PRTChlamydia trachomatis 73Met Lys Lys Leu Leu Lys Ser Val Leu Val Phe Ala Ala Leu Ser Ser 1 5 10 15 Ala Ser Ser Leu Gln Ala Leu Pro Val Gly Asn Pro Ala Glu Pro Ser 20 25 30 Leu Met Ile Asp Gly Ile Leu Trp Glu Gly Phe Gly Gly Asp Pro Cys 35 40 45 Asp Pro Cys Thr Thr Trp Cys Asp Ala Ile Ser Met Arg Met Gly Tyr 50 55 60 Tyr Gly Asp Phe Val Phe Asp Arg Val Leu Lys Thr Asp Val Asn Lys 65 70 75 80 Glu Phe Glu Met Gly Glu Ala Leu Ala Gly Ala Ser Gly Asn Thr Thr 85 90 95 Ser Thr Leu Ser Lys Leu Val Glu Arg Thr Asn Pro Ala Tyr Gly Lys 100 105 110 His Met Gln Asp Ala Glu Met Phe Thr Asn Ala Ala Cys Met Ala Leu 115 120 125 Asn Ile Trp Asp Arg Phe Asp Val Phe Cys Thr Leu Gly Ala Thr Ser 130 135 140 Gly Tyr Leu Arg Gly Asn Ser Ala Ser Phe Asn Leu Val Gly Leu Phe 145 150 155 160 Gly Asp Ser Glu Asn Ala Thr Gln Pro Ala Ala Thr Ser Ile Pro Asn 165 170 175 Val Gln Leu Asn Gln Ser Val Val Glu Leu Tyr Thr Asp Thr Ala Phe 180 185 190 Ala Trp Ser Val Gly Ala Arg Ala Ala Leu Trp Glu Cys Gly Cys Ala 195 200 205 Thr Leu Gly Ala Ser Phe Gln Tyr Ala Gln Ser Lys Pro Lys Val Glu 210 215 220 Glu Leu Asn Val Leu Cys Asn Ala Ala Glu Phe Thr Ile Asn Lys Pro 225 230 235 240 Lys Gly Tyr Val Gly Gln Glu Phe Pro Leu Ala Leu Thr Ala Gly Thr 245 250 255 Asp Ala Ala Thr Gly Thr Lys Asp Ala Ser Ile Asp Tyr His Glu Trp 260 265 270 Gln Ala Ser Leu Ser Leu Ser Tyr Arg Leu Asn Met Phe Thr Pro Tyr 275 280 285 Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ser Asn Thr Ile Arg 290 295 300 Ile Ala Gln Pro Lys Leu Ala Lys Pro Val Val Asp Ile Thr Thr Leu 305 310 315 320 Asn Pro Thr Ile Ala Gly Cys Gly Ser Val Val Ala Ala Asn Ser Glu 325 330 335 Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn Lys 340 345 350 Met Lys Ser Arg Lys Ser Cys Gly Ile Ala Val Gly Thr Thr Ile Val 355 360 365 Asp Ala Asp Lys Tyr Ala Val Thr Val Glu Thr Arg Leu Ile Asp Glu 370 375 380 Arg Ala Ala His Val Asn Ala Gln Phe 385 390 74389PRTChlamydia trachomatis 74Met Lys Lys Leu Leu Lys Ser Val Leu Val Phe Ala Ala Leu Ser Ser 1 5 10 15 Ala Ser Ser Leu Gln Ala Leu Pro Val Gly Asn Pro Ala Glu Pro Ser 20 25 30 Leu Met Ile Asp Gly Ile Leu Trp Glu Gly Phe Gly Gly Asp Pro Cys 35 40 45 Asp Pro Cys Thr Thr Trp Cys Asp Ala Ile Ser Met Arg Met Gly Tyr 50 55 60 Tyr Gly Asp Phe Val Phe Asp Arg Val Leu Lys Thr Asp Val Asn Lys 65 70 75 80 Glu Phe Gln Met Gly Ala Ala Pro Thr Thr Lys

Asp Ile Ala Gly Leu 85 90 95 Glu Asn Asp Pro Thr Thr Asn Val Ala Arg Pro Asn Pro Ala Tyr Gly 100 105 110 Lys His Met Gln Asp Ala Glu Met Phe Thr Asn Ala Ala Tyr Met Ala 115 120 125 Leu Asn Ile Trp Asp Arg Phe Asp Val Phe Cys Thr Leu Gly Ala Thr 130 135 140 Thr Gly Tyr Leu Lys Gly Asn Ser Ala Ser Phe Asn Leu Val Gly Leu 145 150 155 160 Phe Gly Thr Lys Thr Gln Ser Ser Asn Phe Asn Thr Ala Lys Leu Ile 165 170 175 Pro Asn Ala Ala Leu Asn Gln Ala Val Val Glu Leu Tyr Thr Asp Thr 180 185 190 Thr Phe Ala Trp Ser Val Gly Ala Arg Ala Ala Leu Trp Glu Cys Gly 195 200 205 Cys Ala Thr Leu Gly Ala Ser Phe Gln Tyr Ala Gln Ser Lys Pro Lys 210 215 220 Val Glu Glu Leu Asn Val Leu Cys Asn Ala Ser Glu Phe Thr Ile Asn 225 230 235 240 Lys Pro Lys Gly Tyr Val Gly Ala Glu Phe Pro Leu Asp Ile Thr Ala 245 250 255 Gly Thr Glu Ala Ala Thr Gly Thr Lys Asp Ala Ser Ile Asp Tyr His 260 265 270 Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr Arg Leu Asn Met Phe Thr 275 280 285 Pro Tyr Ile Gly Val Lys Trp Ser Arg Val Ser Phe Asp Ala Asp Thr 290 295 300 Ile Arg Ile Ala Gln Pro Lys Leu Ala Glu Ala Ile Leu Asp Val Thr 305 310 315 320 Thr Leu Asn Pro Thr Ile Ala Gly Lys Gly Thr Val Val Ala Ser Gly 325 330 335 Ser Asp Asn Asp Leu Ala Asp Thr Met Gln Ile Val Ser Leu Gln Leu 340 345 350 Asn Lys Met Lys Ser Arg Lys Ser Cys Gly Ile Ala Val Gly Thr Thr 355 360 365 Ile Val Asp Ala Asp Lys Tyr Ala Val Thr Val Glu Thr Arg Leu Ile 370 375 380 Asp Glu Arg Ala Ala 385 75397PRTChlamydia trachomatis 75Met Lys Lys Leu Leu Lys Ser Val Leu Val Phe Ala Ala Leu Ser Ser 1 5 10 15 Ala Ser Ser Leu Gln Ala Leu Pro Val Gly Asn Pro Ala Glu Pro Ser 20 25 30 Leu Met Ile Asp Gly Ile Leu Trp Glu Gly Phe Gly Gly Asp Pro Cys 35 40 45 Asp Pro Cys Thr Thr Trp Cys Asp Ala Ile Ser Met Arg Met Gly Tyr 50 55 60 Tyr Gly Asp Phe Val Phe Asp Arg Val Leu Lys Thr Asp Val Asn Lys 65 70 75 80 Glu Phe Gln Met Gly Ala Ala Pro Thr Thr Ser Asp Val Ala Gly Leu 85 90 95 Gln Asn Asp Pro Thr Thr Asn Val Ala Arg Pro Asn Pro Ala Tyr Gly 100 105 110 Lys His Met Gln Asp Ala Glu Met Phe Thr Asn Ala Ala Tyr Met Ala 115 120 125 Leu Asn Ile Trp Asp Arg Phe Asp Val Phe Cys Thr Leu Gly Ala Thr 130 135 140 Thr Gly Tyr Leu Lys Gly Asn Ser Ala Ser Phe Asn Leu Val Gly Leu 145 150 155 160 Phe Gly Thr Lys Thr Gln Ala Ser Ser Phe Asn Thr Ala Asn Leu Phe 165 170 175 Pro Asn Thr Ala Leu Asn Gln Ala Val Val Glu Leu Tyr Thr Asp Thr 180 185 190 Thr Phe Ala Trp Ser Val Gly Ala Arg Ala Ala Leu Trp Glu Cys Gly 195 200 205 Cys Ala Thr Leu Gly Ala Ser Phe Gln Tyr Ala Gln Ser Lys Pro Lys 210 215 220 Val Glu Glu Leu Asn Val Leu Cys Asn Ala Ser Glu Phe Thr Ile Asn 225 230 235 240 Lys Pro Lys Gly Tyr Val Gly Ala Glu Phe Pro Leu Asp Ile Thr Ala 245 250 255 Gly Thr Glu Ala Ala Thr Gly Thr Lys Asp Ala Ser Ile Asp Tyr His 260 265 270 Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr Arg Leu Asn Met Phe Thr 275 280 285 Pro Tyr Ile Gly Val Lys Trp Ser Arg Val Ser Phe Asp Ala Asp Thr 290 295 300 Ile Arg Ile Ala Gln Pro Lys Leu Ala Glu Ala Ile Leu Asp Val Thr 305 310 315 320 Thr Leu Asn Pro Thr Ile Ala Gly Lys Gly Thr Val Val Ala Ser Gly 325 330 335 Ser Glu Asn Asp Leu Ala Asp Thr Met Gln Ile Val Ser Leu Gln Leu 340 345 350 Asn Lys Met Lys Ser Arg Lys Ser Cys Gly Ile Ala Val Gly Thr Thr 355 360 365 Ile Val Asp Ala Asp Lys Tyr Ala Val Thr Val Glu Thr Arg Leu Ile 370 375 380 Asp Glu Arg Ala Ala His Val Asn Ala Gln Phe Arg Phe 385 390 395 769PRTChlamydia trachomatis 76Thr Thr Leu Asn Pro Thr Ile Ala Gly 1 5 7727PRTChlamydia trachomatis 77Ser Met Arg Val Gly Tyr Tyr Gly Asp Phe Val Phe Asp Arg Val Leu 1 5 10 15 Lys Thr Asp Val Asn Lys Glu Phe Gln Met Gly 20 25 7826PRTChlamydia trachomatis 78Asn Pro Ala Tyr Gly Arg His Met Gln Asp Ala Glu Met Phe Thr Asn 1 5 10 15 Ala Ala Cys Met Ala Leu Asn Ile Trp Asp 20 25 798PRTChlamydia trachomatis 79Ser Ala Thr Ala Ile Phe Asp Thr 1 5 808PRTChlamydia trachomatis 80Leu Val Thr Pro Val Val Asp Ile 1 5 818PRTChlamydia trachomatis 81Leu Ala Lys Pro Val Val Asp Ile 1 5 828PRTChlamydia trachomatis 82Leu Ala Glu Ala Ile Leu Asp Val 1 5 8313PRTChlamydia trachomatis 83Ala Gly Asp Val Lys Thr Gly Ala Glu Gly Gln Leu Gly 1 5 10 8413PRTChlamydia trachomatis 84Ala Gly Asp Val Lys Ala Ser Ala Glu Gly Gln Leu Gly 1 5 10 8514PRTChlamydia trachomatis 85Cys Gly Ser Val Ala Gly Ala Asn Thr Glu Gly Gln Ile Ser 1 5 10 8614PRTChlamydia trachomatis 86Cys Gly Ser Val Val Ala Ala Asn Ser Glu Gly Gln Ile Ser 1 5 10 8713PRTChlamydia trachomatis 87Lys Gly Thr Val Val Ser Ser Ala Glu Asn Glu Leu Ala 1 5 10 8814PRTChlamydia trachomatis 88Lys Gly Thr Val Val Ala Ser Gly Ser Glu Asn Asp Leu Ala 1 5 10 8920PRTChlamydia trachomatis 89Asp Ala Ser Ile Asp Tyr His Glu Trp Gln Ala Ser Leu Ala Leu Ser 1 5 10 15 Tyr Arg Leu Asn 20 9020PRTChlamydia trachomatis 90Ala Ser Leu Ala Leu Ser Tyr Arg Leu Asn Met Phe Thr Pro Tyr Ile 1 5 10 15 Gly Val Lys Trp 20 9120PRTChlamydia trachomatis 91Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp 1 5 10 15 Ala Asp Thr Ile 20 9220PRTChlamydia trachomatis 92Ser Arg Ala Ser Phe Asp Ala Asp Thr Ile Arg Ile Ala Gln Pro Lys 1 5 10 15 Ser Ala Thr Ala 20 9320PRTChlamydia trachomatis 93Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr Thr 1 5 10 15 Leu Asn Pro Thr 20 9420PRTChlamydia trachomatis 94Ile Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp 1 5 10 15 Val Lys Ala Ser 20 9520PRTChlamydia trachomatis 95Ile Ala Gly Ala Gly Asp Val Lys Ala Ser Ala Glu Gly Gln Leu Gly 1 5 10 15 Asp Thr Met Gln 20 9620PRTChlamydia trachomatis 96Ala Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu 1 5 10 15 Asn Lys Met Lys 20 9720PRTChlamydia trachomatis 97Asp Ala Ser Ile Asp Tyr His Glu Trp Gln Ala Ser Leu Ser Leu Ser 1 5 10 15 Tyr Arg Leu Asn 20 9820PRTChlamydia trachomatis 98Ala Ser Leu Ser Leu Ser Tyr Arg Leu Asn Met Phe Thr Pro Tyr Ile 1 5 10 15 Gly Val Lys Trp 20 9920PRTChlamydia trachomatis 99Met Phe Thr Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp 1 5 10 15 Ser Asp Thr Ile 20 10020PRTChlamydia trachomatis 100Ser Arg Ala Ser Phe Asp Ser Asp Thr Ile Arg Ile Ala Gln Pro Arg 1 5 10 15 Leu Val Thr Pro 20 10120PRTChlamydia trachomatis 101Arg Ile Ala Gln Pro Arg Leu Val Thr Pro Val Val Asp Ile Thr Thr 1 5 10 15 Leu Asn Pro Thr 20 10220PRTChlamydia trachomatis 102Val Val Asp Ile Thr Thr Leu Asn Pro Thr Ile Ala Gly Cys Gly Ser 1 5 10 15 Val Ala Gly Ala 20 10321PRTChlamydia trachomatis 103Ile Ala Gly Cys Gly Ser Val Ala Gly Ala Asn Thr Glu Gly Gln Ile 1 5 10 15 Ser Asp Thr Met Gln 20 10420PRTChlamydia trachomatis 104Thr Glu Gly Gln Ile Ser Asp Thr Met Gln Ile Val Ser Leu Gln Leu 1 5 10 15 Asn Lys Met Lys 20 1059PRTChlamydia trachomatis 105Ser Arg Ala Ser Phe Asp Ala Asp Thr 1 5 1069PRTChlamydia trachomatis 106Arg Ala Ser Phe Asp Ala Asp Thr Ile 1 5 1079PRTChlamydia trachomatis 107Ala Ser Phe Asp Ala Asp Thr Ile Arg 1 5 1089PRTChlamydia trachomatis 108Ser Phe Asp Ala Asp Thr Ile Arg Ile 1 5 1099PRTChlamydia trachomatis 109Phe Asp Ala Asp Thr Ile Arg Ile Ala 1 5 1109PRTChlamydia trachomatis 110Asp Ala Asp Thr Ile Arg Ile Ala Gln 1 5 1119PRTChlamydia trachomatis 111Ala Asp Thr Ile Arg Ile Ala Gln Pro 1 5 1129PRTChlamydia trachomatis 112Asp Thr Ile Arg Ile Ala Gln Pro Lys 1 5 1139PRTChlamydia trachomatis 113Thr Ile Arg Ile Ala Gln Pro Lys Ser 1 5 1149PRTChlamydia trachomatis 114Ile Arg Ile Ala Gln Pro Lys Ser Ala 1 5 1159PRTChlamydia trachomatis 115Arg Ile Ala Gln Pro Lys Ser Ala Thr 1 5 1169PRTChlamydia trachomatis 116Ile Ala Gln Pro Lys Ser Ala Thr Ala 1 5 1179PRTChlamydia trachomatis 117Ala Gln Pro Lys Ser Ala Thr Ala Ile 1 5 1189PRTChlamydia trachomatis 118Gln Pro Lys Ser Ala Thr Ala Ile Phe 1 5 1199PRTChlamydia trachomatis 119Pro Lys Ser Ala Thr Ala Ile Phe Asp 1 5 1209PRTChlamydia trachomatis 120Lys Ser Ala Thr Ala Ile Phe Asp Thr 1 5 1219PRTChlamydia trachomatis 121Ser Ala Thr Ala Ile Phe Asp Thr Thr 1 5 1229PRTChlamydia trachomatis 122Ala Thr Ala Ile Phe Asp Thr Thr Thr 1 5 1239PRTChlamydia trachomatis 123Thr Ala Ile Phe Asp Thr Thr Thr Leu 1 5 1249PRTChlamydia trachomatis 124Ala Ile Phe Asp Thr Thr Thr Leu Asn 1 5 1259PRTChlamydia trachomatis 125Ile Phe Asp Thr Thr Thr Leu Asn Pro 1 5 1269PRTChlamydia trachomatis 126Phe Asp Thr Thr Thr Leu Asn Pro Thr 1 5 1279PRTChlamydia trachomatis 127Asp Thr Thr Thr Leu Asn Pro Thr Ile 1 5 1289PRTChlamydia trachomatis 128Thr Thr Thr Leu Asn Pro Thr Ile Ala 1 5 1299PRTChlamydia trachomatis 129Thr Leu Asn Pro Thr Ile Ala Gly Ala 1 5 1309PRTChlamydia trachomatis 130Leu Asn Pro Thr Ile Ala Gly Ala Gly 1 5 1319PRTChlamydia trachomatis 131Asn Pro Thr Ile Ala Gly Ala Gly Asp 1 5 1329PRTChlamydia trachomatis 132Pro Thr Ile Ala Gly Ala Gly Asp Val 1 5 1339PRTChlamydia trachomatis 133Thr Ile Ala Gly Ala Gly Asp Val Lys 1 5 1349PRTChlamydia trachomatis 134Ile Ala Gly Ala Gly Asp Val Lys Thr 1 5 1359PRTChlamydia trachomatis 135Ala Gly Ala Gly Asp Val Lys Thr Gly 1 5 1369PRTChlamydia trachomatis 136Gly Ala Gly Asp Val Lys Thr Gly Ala 1 5 1379PRTChlamydia trachomatis 137Ala Gly Asp Val Lys Thr Gly Ala Glu 1 5 1389PRTChlamydia trachomatis 138Gly Asp Val Lys Thr Gly Ala Glu Gly 1 5 1399PRTChlamydia trachomatis 139Asp Val Lys Thr Gly Ala Glu Gly Gln 1 5 1409PRTChlamydia trachomatis 140Val Lys Thr Gly Ala Glu Gly Gln Leu 1 5 1419PRTChlamydia trachomatis 141Lys Thr Gly Ala Glu Gly Gln Leu Gly 1 5 1429PRTChlamydia trachomatis 142Thr Gly Ala Glu Gly Gln Leu Gly Asp 1 5 1439PRTChlamydia trachomatis 143Gly Ala Glu Gly Gln Leu Gly Asp Thr 1 5 1449PRTChlamydia trachomatis 144Ala Glu Gly Gln Leu Gly Asp Thr Met 1 5 1459PRTChlamydia trachomatis 145Glu Gly Gln Leu Gly Asp Thr Met Gln 1 5 1469PRTChlamydia trachomatis 146Gly Gln Leu Gly Asp Thr Met Gln Ile 1 5 1479PRTChlamydia trachomatis 147Gln Leu Gly Asp Thr Met Gln Ile Val 1 5 1489PRTChlamydia trachomatis 148Leu Gly Asp Thr Met Gln Ile Val Ser 1 5 1499PRTChlamydia trachomatis 149Ser Arg Ala Ser Phe Asp Ser Asp Thr 1 5 1509PRTChlamydia trachomatis 150Arg Ala Ser Phe Asp Ser Asp Thr Ile 1 5 1519PRTChlamydia trachomatis 151Ala Ser Phe Asp Ser Asp Thr Ile Arg 1 5 1529PRTChlamydia trachomatis 152Ser Phe Asp Ser Asp Thr Ile Arg Ile 1 5 1539PRTChlamydia trachomatis 153Phe Asp Ser Asp Thr Ile Arg Ile Ala 1 5 1549PRTChlamydia trachomatis 154Asp Ser Asp Thr Ile Arg Ile Ala Gln 1 5 1559PRTChlamydia trachomatis 155Ser Asp Thr Ile Arg Ile Ala Gln Pro 1 5 1569PRTChlamydia trachomatis 156Asp Thr Ile Arg Ile Ala Gln Pro Arg 1 5 1579PRTChlamydia trachomatis 157Thr Ile Arg Ile Ala Gln Pro Arg Leu 1 5 1589PRTChlamydia trachomatis 158Ile Arg Ile Ala Gln Pro Arg Leu Val 1 5 1599PRTChlamydia trachomatis 159Arg Ile Ala Gln Pro Arg Leu Val Thr 1 5 1609PRTChlamydia trachomatis 160Ile Ala Gln Pro Arg Leu Val Thr Pro 1 5 1619PRTChlamydia trachomatis 161Ala Gln Pro Arg Leu Val Thr Pro Val 1 5 1629PRTChlamydia trachomatis 162Gln Pro Arg Leu Val Thr Pro Val Val 1 5 1639PRTChlamydia trachomatis 163Pro Arg Leu Val Thr Pro Val Val Asp 1 5 1649PRTChlamydia trachomatis 164Arg Leu Val Thr Pro Val Val Asp Ile 1 5 1659PRTChlamydia trachomatis 165Leu Val Thr Pro Val Val Asp Ile Thr 1 5 1669PRTChlamydia trachomatis 166Val Thr Pro Val Val Asp Ile Thr Thr 1 5 1679PRTChlamydia trachomatis 167Thr Pro Val Val Asp Ile Thr Thr Leu 1 5 1689PRTChlamydia trachomatis 168Pro Val Val Asp Ile Thr Thr Leu Asn 1 5 1699PRTChlamydia trachomatis 169Val Val Asp Ile Thr Thr Leu Asn Pro 1 5 1709PRTChlamydia trachomatis 170Val Asp Ile Thr Thr Leu Asn Pro Thr 1 5 1719PRTChlamydia trachomatis 171Asp Ile Thr Thr Leu Asn Pro Thr Ile 1 5 1729PRTChlamydia trachomatis 172Ile Thr Thr Leu Asn Pro Thr Ile Ala 1 5 1739PRTChlamydia trachomatis 173Thr Leu Asn Pro Thr Ile Ala Gly Cys 1 5 1749PRTChlamydia trachomatis 174Leu Asn Pro Thr Ile Ala Gly Cys Gly 1 5 1759PRTChlamydia trachomatis 175Asn Pro Thr Ile Ala Gly Cys Gly Ser 1

5 1769PRTChlamydia trachomatis 176Pro Thr Ile Ala Gly Cys Gly Ser Val 1 5 1779PRTChlamydia trachomatis 177Thr Ile Ala Gly Cys Gly Ser Val Ala 1 5 1789PRTChlamydia trachomatis 178Ile Ala Gly Cys Gly Ser Val Ala Gly 1 5 1799PRTChlamydia trachomatis 179Ala Gly Cys Gly Ser Val Ala Gly Ala 1 5 1809PRTChlamydia trachomatis 180Gly Cys Gly Ser Val Ala Gly Ala Asn 1 5 1819PRTChlamydia trachomatis 181Cys Gly Ser Val Ala Gly Ala Asn Thr 1 5 1829PRTChlamydia trachomatis 182Gly Ser Val Ala Gly Ala Asn Thr Glu 1 5 1839PRTChlamydia trachomatis 183Ser Val Ala Gly Ala Asn Thr Glu Gly 1 5 1849PRTChlamydia trachomatis 184Val Ala Gly Ala Asn Thr Glu Gly Gln 1 5 1859PRTChlamydia trachomatis 185Ala Gly Ala Asn Thr Glu Gly Gln Ile 1 5 1869PRTChlamydia trachomatis 186Gly Ala Asn Thr Glu Gly Gln Ile Ser 1 5 1879PRTChlamydia trachomatis 187Ala Asn Thr Glu Gly Gln Ile Ser Asp 1 5 1889PRTChlamydia trachomatis 188Asn Thr Glu Gly Gln Ile Ser Asp Thr 1 5 1899PRTChlamydia trachomatis 189Thr Glu Gly Gln Ile Ser Asp Thr Met 1 5 1909PRTChlamydia trachomatis 190Glu Gly Gln Ile Ser Asp Thr Met Gln 1 5 1919PRTChlamydia trachomatis 191Gly Gln Ile Ser Asp Thr Met Gln Ile 1 5 1929PRTChlamydia trachomatis 192Gln Ile Ser Asp Thr Met Gln Ile Val 1 5 1939PRTChlamydia trachomatis 193Ile Ser Asp Thr Met Gln Ile Val Ser 1 5 19417PRTChlamydia trachomatis 194Phe Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val 1 5 10 15 Lys 1959PRTChlamydia trachomatis 195Asn Met Phe Thr Pro Tyr Ile Gly Val 1 5 1969PRTChlamydia trachomatis 196Met Phe Thr Pro Tyr Ile Gly Val Lys 1 5 1979PRTChlamydia trachomatis 197Phe Thr Pro Tyr Ile Gly Val Lys Trp 1 5 1989PRTChlamydia trachomatis 198Thr Pro Tyr Ile Gly Val Lys Trp Ser 1 5 1999PRTChlamydia trachomatis 199Pro Tyr Ile Gly Val Lys Trp Ser Arg 1 5 2009PRTChlamydia trachomatis 200Tyr Ile Gly Val Lys Trp Ser Arg Ala 1 5 2019PRTChlamydia trachomatis 201Ile Gly Val Lys Trp Ser Arg Ala Ser 1 5 2029PRTChlamydia trachomatis 202Gly Val Lys Trp Ser Arg Ala Ser Phe 1 5 2039PRTChlamydia trachomatis 203Val Lys Trp Ser Arg Ala Ser Phe Asp 1 5 2049PRTChlamydia trachomatis 204Lys Trp Ser Arg Ala Ser Phe Asp Ala 1 5 2059PRTChlamydia trachomatis 205Trp Ser Arg Ala Ser Phe Asp Ala Asp 1 5 2069PRTChlamydia trachomatis 206Ser Arg Ala Ser Phe Asp Ala Asp Thr 1 5 2079PRTChlamydia trachomatis 207Arg Ala Ser Phe Asp Ala Asp Thr Ile 1 5 2089PRTChlamydia trachomatis 208Ala Ser Phe Asp Ala Asp Thr Ile Arg 1 5 2099PRTChlamydia trachomatis 209Ser Phe Asp Ala Asp Thr Ile Arg Ile 1 5 2109PRTChlamydia trachomatis 210Phe Asp Ala Asp Thr Ile Arg Ile Ala 1 5 2119PRTChlamydia trachomatis 211Asp Ala Asp Thr Ile Arg Ile Ala Gln 1 5 2129PRTChlamydia trachomatis 212Ala Asp Thr Ile Arg Ile Ala Gln Pro 1 5 2139PRTChlamydia trachomatis 213Asp Thr Ile Arg Ile Ala Gln Pro Lys 1 5 2149PRTChlamydia trachomatis 214Thr Ile Arg Ile Ala Gln Pro Lys Ser 1 5 2159PRTChlamydia trachomatis 215Ile Arg Ile Ala Gln Pro Lys Ser Ala 1 5 2169PRTChlamydia trachomatis 216Arg Ile Ala Gln Pro Lys Ser Ala Thr 1 5 2179PRTChlamydia trachomatis 217Ile Ala Gln Pro Lys Ser Ala Thr Ala 1 5 2189PRTChlamydia trachomatis 218Ala Gln Pro Lys Ser Ala Thr Ala Ile 1 5 2199PRTChlamydia trachomatis 219Gln Pro Lys Ser Ala Thr Ala Ile Phe 1 5 2209PRTChlamydia trachomatis 220Pro Lys Ser Ala Thr Ala Ile Phe Asp 1 5 2219PRTChlamydia trachomatis 221Lys Ser Ala Thr Ala Ile Phe Asp Thr 1 5 2229PRTChlamydia trachomatis 222Ser Ala Thr Ala Ile Phe Asp Thr Thr 1 5 2239PRTChlamydia trachomatis 223Ala Thr Ala Ile Phe Asp Thr Thr Thr 1 5 2249PRTChlamydia trachomatis 224Thr Ala Ile Phe Asp Thr Thr Thr Leu 1 5 2259PRTChlamydia trachomatis 225Ala Ile Phe Asp Thr Thr Thr Leu Asn 1 5 2269PRTChlamydia trachomatis 226Ile Phe Asp Thr Thr Thr Leu Asn Pro 1 5 2279PRTChlamydia trachomatis 227Phe Asp Thr Thr Thr Leu Asn Pro Thr 1 5 2289PRTChlamydia trachomatis 228Asp Thr Thr Thr Leu Asn Pro Thr Ile 1 5 2299PRTChlamydia trachomatis 229Thr Thr Thr Leu Asn Pro Thr Ile Ala 1 5 2309PRTChlamydia trachomatis 230Thr Thr Leu Asn Pro Thr Ile Ala Gly 1 5 2319PRTChlamydia trachomatis 231Thr Leu Asn Pro Thr Ile Ala Gly Ala 1 5 2329PRTChlamydia trachomatis 232Leu Asn Pro Thr Ile Ala Gly Ala Gly 1 5 2339PRTChlamydia trachomatis 233Asn Pro Thr Ile Ala Gly Ala Gly Asp 1 5 2349PRTChlamydia trachomatis 234Pro Thr Ile Ala Gly Ala Gly Asp Val 1 5 2359PRTChlamydia trachomatis 235Thr Ile Ala Gly Ala Gly Asp Val Lys 1 5 2369PRTChlamydia trachomatis 236Ile Ala Gly Ala Gly Asp Val Lys Thr 1 5 2379PRTChlamydia trachomatis 237Ala Gly Ala Gly Asp Val Lys Thr Gly 1 5 2389PRTChlamydia trachomatis 238Gly Ala Gly Asp Val Lys Thr Gly Ala 1 5 2399PRTChlamydia trachomatis 239Ala Gly Asp Val Lys Thr Gly Ala Glu 1 5 2409PRTChlamydia trachomatis 240Gly Asp Val Lys Thr Gly Ala Glu Gly 1 5 2419PRTChlamydia trachomatis 241Asp Val Lys Thr Gly Ala Glu Gly Gln 1 5 2429PRTChlamydia trachomatis 242Val Lys Thr Gly Ala Glu Gly Gln Leu 1 5 2439PRTChlamydia trachomatis 243Lys Thr Gly Ala Glu Gly Gln Leu Gly 1 5 2449PRTChlamydia trachomatis 244Thr Gly Ala Glu Gly Gln Leu Gly Asp 1 5 2459PRTChlamydia trachomatis 245Gly Ala Glu Gly Gln Leu Gly Asp Thr 1 5 2469PRTChlamydia trachomatis 246Ala Glu Gly Gln Leu Gly Asp Thr Met 1 5 2479PRTChlamydia trachomatis 247Glu Gly Gln Leu Gly Asp Thr Met Gln 1 5 2489PRTChlamydia trachomatis 248Gly Gln Leu Gly Asp Leu Met Gln Ile 1 5 2499PRTChlamydia trachomatis 249Gln Leu Gly Asp Thr Met Gln Ile Val 1 5 2509PRTChlamydia trachomatis 250Leu Gly Asp Thr Met Gln Ile Val Ser 1 5 2519PRTChlamydia trachomatis 251Gly Asp Thr Met Gln Ile Val Ser Leu 1 5 2529PRTChlamydia trachomatis 252Asp Thr Met Gln Ile Val Ser Leu Gln 1 5 2539PRTChlamydia trachomatis 253Thr Met Gln Ile Val Ser Leu Gln Leu 1 5 2549PRTChlamydia trachomatis 254Met Gln Ile Val Ser Leu Gln Leu Asn 1 5 25580PRTChlamydia trachomatis 255Glu Trp Gln Ala Ser Leu Ala Leu Ser Tyr Arg Leu Asn Met Phe Thr 1 5 10 15 Pro Tyr Ile Gly Val Lys Trp Ser Arg Ala Ser Phe Asp Ala Asp Thr 20 25 30 Ile Arg Ile Ala Gln Pro Lys Ser Ala Thr Ala Ile Phe Asp Thr Thr 35 40 45 Thr Leu Asn Pro Thr Ile Ala Gly Ala Gly Asp Val Lys Thr Gly Ala 50 55 60 Glu Gly Gln Leu Gly Asp Thr Met Gln Ile Val Ser Leu Gln Leu Asn 65 70 75 80

Claims

1. A polypeptide comprising a) an amino acid sequence comprising one or more surface exposed fragments of the same outer membrane protein expressed in a serotype of Chlamydia sp.; and b) two or more additional amino acid sequences which is either the same sequence as defined in a) or is the corresponding surface exposed fragments from a variant of said outer membrane protein expressed in a serotype of Chlamydia sp., which is different from the serotype in a).

2. A polypeptide according to claim 1, comprising three or more different amino acid sequences, where said amino acid sequences each comprise one or more surface exposed fragments from different variants of the same outer membrane protein that varies in different Chlamydia sp. serotypes, said amino acid sequences derived from different Chlamydia sp. serotypes.

3. A polypeptide according to claim 1, comprising three or more repetitions of an amino acid sequence, where said amino acid sequence comprises one or more surface exposed fragments of the same outer membrane protein that varies in different Chlamydia sp. serotypes, said amino acid sequences derived from the same Chlamydia sp. serotype.

4. A polypeptide according to claim 1, wherein the outer membrane protein is MOMP from any serotype.

5. A polypeptide according to claim 4, wherein the outer membrane protein is MOMP from serotype D, E, F, G, Ia or J of Chlamydia trachomatis or C. pneumoniae.

6. A polypeptide according to claim 4, comprising one or more of the variable domains selected from 1, 2, 3, 4 of MOMP.

7. A polypeptide according to claim 6, where the amino acid sequences are linearized.

8. A polypeptide according to claim 6, wherein the amino acid sequences comprising the variable domains 4 (VD4) of MOMP are placed next to each other or are spaced with a linker.

9. A polypeptide according to claim 8, comprising an amino acid sequence defined in formula I: xx₁-VD4-xx₂ (Formula I) wherein VD4 is independently selected from SEQ ID NO. 15-20 or an amino acid sequence which has at least 80% sequence identity with SEQ ID NO 15-20, and xx₁ consists of i) the amino acid sequence EWQASLALSYRLNMFTPYIGVKWSRASFDADTIRIAQPK (SEQ ID NO 21) or ii) a subsequence of the amino acid sequence in i) said subsequence comprising 1-38 amino acid residues, starting with the C-terminal K in the amino acid sequence in i) and xx₂ consists of iii) the amino acid sequence DTMQIVSLQLNKMKSRKSCGIAVGTTIVDA (SEQ ID NO 22) or iv) a subsequence of the amino acid sequence in iii) said subsequence comprising 1-29 amino acid residues, starting with the N-terminal D in the amino acid sequence in iii).

10. A polypeptide according to claim 8, where said sequences are selected from SEQ ID NO. 23-28 and 49-59.

11. A polypeptide according to claim 1, further comprising a fragment comprising the variable domains 1 (VD1) of MOMP and wherein the amino acid sequences comprising VD1 of MOMP are placed next to each other or are spaced with a linker.

12. A polypeptide according to claim 11 comprising an amino acid sequence defined in formula II: yy₁-VD1-yy₂ (Formula II) wherein VD1 is independently selected from SEQ ID NO 1-6 or an amino acid sequence which has at least 80% sequence identity with SEQ ID NO 1-6, and yy₁ consists of v) the amino acid sequence DAISMRVGYYGDFVFDRVLKTDVNKEFQMG (SEQ ID NO 7) or vi) a subsequence of the amino acid sequence in v) said subsequence comprising 1-30 amino acid residues, starting with the C-terminal G in the amino acid sequence in v) and yy₂ consists of vii) the amino acid sequence NPAYGRHMQDAEMFTNAA (SEQ ID NO 8) or viii) a subsequence of the amino acid sequence in vii) said subsequence comprising 1-18 amino acid residues, starting with the N-terminal N in the amino acid sequence in vii).

13. A polypeptide according to claim 11, where said sequences are selected from SEQ ID NO. 9-14 and 45-48.

14. A polypeptide according to claim 1, further comprising a fragment comprising the variable domains 2 (VD2) of MOMP and wherein the amino acid sequences comprising VD2 of MOMP are placed next to each other or are spaced with a linker.

15. A polypeptide according to claim 14 comprising an amino acid sequence defined in formula III: zz₁-VD2-zz₂ (Formula III) wherein VD2 is independently selected from SEQ ID NO 29-34 or an amino acid sequence which has at least 80% sequence identity with SEQ ID NO 29-34, and zz₁ consists of ix) the amino acid sequence TLGATSGYLKGNSASFNLVGLFG (SEQ ID NO 35) or x) a subsequence of the amino acid sequence in ix) said subsequence comprising 1-23 amino acid residues, starting with the C-terminal G in the amino acid sequence in ix) and xx₂ consists of xi) the amino acid sequence WELYTDTTFAWSVGARAALWE (SEQ ID NO 36) or xii) a subsequence of the amino acid sequence in xi) said subsequence comprising 1-22 amino acid residues, starting with the N-terminal V in the amino acid sequence in xi).

16. A polypeptide according to claim 1, additionally comprising a fragment comprising the variable domains 3 (VD3) of MOMP and wherein the amino acid sequences comprising VD3 of MOMP are placed next to each other or are spaced with a linker.

17. A polypeptide according to claim 16 comprising an amino acid sequence defined in formula IV: qq₁-VD3-qq₂ (Formula IV) wherein VD3 is independently selected from SEQ ID NO 37-42 or an amino acid sequence which has at least 80% sequence identity with SEQ ID NO 37-42, and qq₁ consists of xiii) the amino acid sequence ATLGASFQYAQSKPKVEELNVLCNAAEFTINKPKGYVG (SEQ ID NO 43) or xiv) a subsequence of the amino acid sequence in xiii) said subsequence comprising 1-22 amino acid residues, starting with the C-terminal G in the amino acid sequence in xiii) and qq₂ consists of xv) the amino acid sequence TGTKDASIDYHEWQASLALSYRLNMFTPYIGVKWS (SEQ ID NO 44) or xvi) a subsequence of the amino acid sequence in xv) said subsequence comprising 1-35 amino acid residues, starting with the N-terminal T in the amino acid sequence in xv).

18. A polypeptide according to claim 1, further comprising a moiety that facilitate export of the polypeptide when produced recombinantly, a moiety that facilitate purification of the fusion protein, or a moiety which enhances immunogenicity.

19. A polypeptide according to claim 18, wherein the enhancer of immunogenicity is an additional T-cell target which is chosen from a Ct antigen such as CT043, CT004, CT414, CT681 or part hereof.

20. A polypeptide according to claim 19, where said sequences are selected from SEQ ID NO 60-68.

Images