We evaluated 7 C. muridarum ORFs for their ability to induce protection against chlamydial infection in a mouse intravaginal infection model. These antigens, although encoded in C. muridarum genome, are transcriptionally regulated by a cryptic plasmid that is known to contribute to C. muridarum pathogenesis. Of the 7 plasmid-regulated ORFs, the chlamydial glycogen phosphorylase or GlgP, when delivered into mice intramuscularly, induced the most pronounced protective immunity against C. muridarum intravaginal infection. The GlgP-immunized mice displayed a significant reduction in vaginal shedding of live organisms on day 14 after infection. The protection correlated well with a robust C. muridarum-specific antibody and a Th1-dominant T cell responses, which significantly reduced the severity but not overall incidence of hydrosalpinx. The GlgP-induced partial protection against upper genital tract pathology suggests that GlgP may be considered a component for a multi-subunit vaccine. These results have demonstrated that intramuscular immunization of mice with purified proteins can be used to identify vaccine antigens for preventing intravaginal infection with C. trachomatis in humans.
References
[1]
Sherman KJ, Daling JR, Stergachis A, Weiss NS, Foy HM, et al. (1990) Sexually transmitted diseases and tubal pregnancy. Sex Transm Dis 17: 115–121.
[2]
Peterman TA, Tian LH, Metcalf CA, Satterwhite CL, Malotte CK, et al. (2006) High incidence of new sexually transmitted infections in the year following a sexually transmitted infection: a case for rescreening. Ann Intern Med 145: 564–572.
[3]
Centers for Disease Control and Prevention CServices USDoHaH, editor (November 2009) Sexually Transmitted Disease Surveillance, 2008. Atlanta, GA: http://www.cdc.gov/std/stats08/toc.htm.
[4]
Kinnunen AH, Surcel HM, Lehtinen M, Karhukorpi J, Tiitinen A, et al. (2002) HLA DQ alleles and interleukin-10 polymorphism associated with Chlamydia trachomatis-related tubal factor infertility: a case-control study. Hum Reprod 17: 2073–2078.
[5]
Rodgers AK, Wang J, Zhang Y, Holden A, Berryhill B, et al. (2010) Association of tubal factor infertility with elevated antibodies to Chlamydia trachomatis caseinolytic protease P. Am J Obstet Gynecol 203: 494 e497–494 e414.
[6]
Rockey DD, Wang J, Lei L, Zhong G (2009) Chlamydia vaccine candidates and tools for chlamydial antigen discovery. Expert Rev Vaccines 8: 1365–1377.
[7]
Grayston JT, Woolridge RL, Wang S (1962) Trachoma vaccine studies on Taiwan. Ann N Y Acad Sci 98: 352–367.
[8]
Zhong G, Lei L, S. G, Lu C, Qi M, et al. (2011) Chlamydia-secreted proteins in chlamydial interactions with host cells. Current Chemical Biology 5: 9.
[9]
Qi M, Lei L, Gong S, Liu Q, Delisa MP, et al. (2011) Chlamydia trachomatis Secretion of an Immunodominant Hypothetical Protein (CT795) into Host Cell Cytoplasm. J Bacteriol 193: 2498–2509.
[10]
Lei L, Qi M, Budrys N, Schenken R, Zhong G (2011) Localization of Chlamydia trachomatis hypothetical protein CT311 in host cell cytoplasm. Microb Pathog 51: 101–109.
[11]
Li Z, Chen C, Chen D, Wu Y, Zhong Y, et al. (2008) Characterization of fifty putative inclusion membrane proteins encoded in the Chlamydia trachomatis genome. Infect Immun 76: 2746–2757.
[12]
Pal S, Peterson EM, de la Maza LM (2005) Vaccination with the Chlamydia trachomatis major outer membrane protein can elicit an immune response as protective as that resulting from inoculation with live bacteria. Infect Immun 73: 8153–8160.
[13]
Pal S, Barnhart KM, Wei Q, Abai AM, Peterson EM, et al. (1999) 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 17: 459–465.
[14]
Zhang D, Yang X, Berry J, Shen C, McClarty G, et al. (1997) DNA vaccination with the major outer-membrane protein gene induces acquired immunity to Chlamydia trachomatis (mouse pneumonitis) infection. J Infect Dis 176: 1035–1040.
[15]
Morrison RP, Caldwell HD (2002) Immunity to murine chlamydial genital infection. Infect Immun 70: 2741–2751.
[16]
Morrison RP, Feilzer K, Tumas DB (1995) Gene knockout mice establish a primary protective role for major histocompatibility complex class II-restricted responses in Chlamydia trachomatis genital tract infection. Infect Immun 63: 4661–4668.
[17]
Stephens RS (2003) The cellular paradigm of chlamydial pathogenesis. Trends Microbiol 11: 44–51.
[18]
Zhong G (2009) Killing me softly: chlamydial use of proteolysis for evading host defenses. Trends Microbiol 17: 467–474.
[19]
Cheng W, Shivshankar P, Li Z, Chen L, Yeh IT, et al. (2008) Caspase-1 contributes to Chlamydia trachomatis-induced upper urogenital tract inflammatory pathologies without affecting the course of infection. Infect Immun 76: 515–522.
[20]
Chen L, Lei L, Chang X, Li Z, Lu C, et al. (2010) Mice deficient in MyD88 Develop a Th2-dominant response and severe pathology in the upper genital tract following Chlamydia muridarum infection. J Immunol 184: 2602–2610.
[21]
O'Connell CM, Ingalls RR, Andrews CW Jr, Scurlock AM, Darville T (2007) Plasmid-deficient Chlamydia muridarum fail to induce immune pathology and protect against oviduct disease. J Immunol 179: 4027–4034.
[22]
Pal S, Davis HL, Peterson EM, de la Maza LM (2002) Immunization with the Chlamydia trachomatis mouse pneumonitis major outer membrane protein by use of CpG oligodeoxynucleotides as an adjuvant induces a protective immune response against an intranasal chlamydial challenge. Infect Immun 70: 4812–4817.
[23]
Morrison SG, Morrison RP (2005) A predominant role for antibody in acquired immunity to chlamydial genital tract reinfection. J Immunol 175: 7536–7542.
[24]
Olivares-Zavaleta N, Whitmire W, Gardner D, Caldwell HD (2010) Immunization with the attenuated plasmidless Chlamydia trachomatis L2(25667R) strain provides partial protection in a murine model of female genitourinary tract infection. Vaccine 28: 1454–1462.
[25]
Carlson JH, Whitmire WM, Crane DD, Wicke L, Virtaneva K, et al. (2008) The Chlamydia trachomatis plasmid is a transcriptional regulator of chromosomal genes and a virulence factor. Infect Immun 76: 2273–2283.
[26]
Wang J, Zhang Y, Lu C, Lei L, Yu P, et al. (2010) A Genome-Wide Profiling of the Humoral Immune Response to Chlamydia trachomatis Infection Reveals Vaccine Candidate Antigens Expressed in Humans. J Immunol 185: 1670–1680.
[27]
Wang J, Chen L, Chen F, Zhang X, Zhang Y, et al. (2009) A chlamydial type III-secreted effector protein (Tarp) is predominantly recognized by antibodies from humans infected with Chlamydia trachomatis and induces protective immunity against upper genital tract pathologies in mice. Vaccine 27: 2967–2980.
[28]
Karunakaran KP, Rey-Ladino J, Stoynov N, Berg K, Shen C, et al. (2008) Immunoproteomic discovery of novel T cell antigens from the obligate intracellular pathogen Chlamydia. J Immunol 180: 2459–2465.
[29]
Eissenberg LG, Wyrick PB (1981) Inhibition of phagolysosome fusion is localized to Chlamydia psittaci-laden vacuoles. Infect Immun 32: 889–896.
[30]
Sharma J, Zhong Y, Dong F, Piper JM, Wang G, et al. (2006) Profiling of human antibody responses to Chlamydia trachomatis urogenital tract infection using microplates arrayed with 156 chlamydial fusion proteins. Infect Immun 74: 1490–1499.
[31]
Strandell A, Lindhard A, Waldenstrom U, Thorburn J, Janson PO, et al. (1999) Hydrosalpinx and IVF outcome: a prospective, randomized multicentre trial in Scandinavia on salpingectomy prior to IVF. Hum Reprod 14: 2762–2769.
[32]
Strandell A, Thorburn J, Wallin A (2004) The presence of cytokines and growth factors in hydrosalpingeal fluid. J Assist Reprod Genet 21: 241–247.
[33]
Strandell A (2000) The influence of hydrosalpinx on IVF and embryo transfer: a review. Hum Reprod Update 6: 387–395.
[34]
Johnson N, van Voorst S, Sowter MC, Strandell A, Mol BW (2010) Surgical treatment for tubal disease in women due to undergo in vitro fertilisation. Cochrane Database Syst Rev CD002125.
[35]
Xiao Y, Zhong Y, Su H, Zhou Z, Chiao P, et al. (2005) NF-kappa B activation is not required for Chlamydia trachomatis inhibition of host epithelial cell apoptosis. J Immunol 174: 1701–1708.
[36]
Zhong G, Fan P, Ji H, Dong F, Huang Y (2001) Identification of a chlamydial protease-like activity factor responsible for the degradation of host transcription factors. J Exp Med 193: 935–942.
[37]
Fan T, Lu H, Hu H, Shi L, McClarty GA, et al. (1998) Inhibition of apoptosis in chlamydia-infected cells: blockade of mitochondrial cytochrome c release and caspase activation. J Exp Med 187: 487–496.
[38]
Greene W, Xiao Y, Huang Y, McClarty G, Zhong G (2004) Chlamydia-infected cells continue to undergo mitosis and resist induction of apoptosis. Infect Immun 72: 451–460.
[39]
Cheng W, Shivshankar P, Zhong Y, Chen D, Li Z, et al. (2008) Intracellular interleukin-1alpha mediates interleukin-8 production induced by Chlamydia trachomatis infection via a mechanism independent of type I interleukin-1 receptor. Infect Immun 76: 942–951.
[40]
Zhong G, Reis e Sousa C, Germain RN (1997) Production, specificity, and functionality of monoclonal antibodies to specific peptide-major histocompatibility complex class II complexes formed by processing of exogenous protein. Proc Natl Acad Sci U S A 94: 13856–13861.
[41]
Zhong G, Reis e Sousa C, Germain RN (1997) Antigen-unspecific B cells and lymphoid dendritic cells both show extensive surface expression of processed antigen-major histocompatibility complex class II complexes after soluble protein exposure in vivo or in vitro. J Exp Med 186: 673–682.
