Identifying relevant animal challenge models adds to the complexity of human vaccine development. Murine challenge models have been the most utilized animal model for Chlamydia trachomatis vaccine development. The question arises as to whether the C. trachomatis or C. muridarum pre-clinical model is optimal. We compared C.muridarum and C. trachomatis intravaginal challenge models in a combined total of seventy-five studies evaluating potential vaccine candidates. In 100% (42/42) of C.muridarum studies, mice immunized with Chlamydia elementary bodies (EB) demonstrated a significant reduction in urogenital bacterial shedding as measured by qPCR (p < 0.05) compared to adjuvant-control-immunized mice. Significant reduction in urogenital shedding was observed for EB-immunized groups in only 82% (27/33) of C. trachomatis studies. We have evaluated proposed vaccine antigens in both models and observed immunization with Chlamydia major outer membrane protein (MOMP) vaccine formulations to be protective (p < 0.05) in both models, immunization with polymorphic membrane protein serovar D (PmpD) p73 passenger domain was protective only in the C. trachomatis model, and immunization with PmpD p82 translocator domain was not protective in either model. We also observed in both models that depletion of CD4+ T-cells in MOMP-immunized mice resulted in diminished protective immunity but animals were still able to reduce the infection level. In contrast, mice immunized with live EBs by intraperitoneal route did not require CD4+ T-cells to resolve urogenital infection from intravaginal challenge in either model. Overall, we have found the C. muridarum model to be a more robust, reliable, and reproducible model for vaccine antigen discovery.
References
[1]
Centers for Disease Control and Prevention (2014) Sexually Transmitted Disease Surveillance 2013. US Department of Health and Human Services, Atlanta.
[2]
Geisler, W.M. (2015) Diagnosis and Management of Uncomplicated Chlamydia trachomatis Infections in Adolescents and Adults: Summary of Evidence Reviewed for the 2010 Centers for Disease Control and Prevention Sexually Transmitted Diseases Treatment Guidelines. Clinical Infectious Diseases, 53, S92-S98.
https://doi.org/10.1093/cid/cir698
[3]
Newman, L., Rowley, J., Vander Hoorn, S., Wijesooriya, N.S., Unemo, M., Low, N., et al. (2015) Global Estimates of the Prevalence and Incidence of Four Curable Sexually Transmitted Infections in 2012 Based on Systematic Review and Global Reporting. PLoS ONE, 10, e0143304. https://doi.org/10.1371/journal.pone.0143304
[4]
Farley, T.A., Cohen, D.A. and Elkins, W. (2003) Asymptomatic Sexually Transmitted Diseases: The Case for Screening. Preventive Medicine, 36, 502-509.
https://doi.org/10.1016/S0091-7435(02)00058-0
[5]
Detels, R., Green, A.M., Klausner, J.D., Katzenstein, D., Gaydos, C., Handsfield, H., et al. (2011) The Incidence and Correlates of Symptomatic and Asymptomatic Chlamydia trachomatis and Neisseria gonorrhoeae Infections in Selected Populations in Five Countries. Sexually Transmitted Diseases, 38, 503-509.
https://doi.org/10.1097/OLQ.0b013e318206c288
[6]
Price, M.J., Ades, A.E., Soldan, K., Welton, N.J., Macleod, J., Simms, I., et al. (2016) The Natural History of Chlamydia trachomatis Infection in Women: A Multi-Parameter Evidence Synthesis. Health Technology Assessment, 20, 1-250.
https://doi.org/10.3310/hta20220
[7]
Nigg, C. (1942) An Unidentified Virus Which Produces Pneumonia and Systemic Infection in Mice. Science, 95, 49-50. https://doi.org/10.1126/science.95.2454.49-a
[8]
Nigg, C. and Eaton, M.D. (1944) Isolation from Normal Mice of a Pneumotropic Virus Which Forms Elementary Bodies. Journal of Experimental Medicine, 79, 497-510. https://doi.org/10.1084/jem.79.5.497
[9]
Morrison, R.P., Feilzer, K. and Tumas, D.B. (1995) Gene Knockout Mice Establish a Primary Protective Role for Major Histocompatibility Complex Class II-Restricted Responses in Chlamydia trachomatis Genital Tract Infection. Infection and Immunity, 63, 4661-4668.
[10]
Riley, M.M., Zurenski, M.A., Frazer, L.C., O’Connell, C.M., Andrews, C.W., Jr., Mintus, M., et al. (2012) The Recall Response Induced by Genital Challenge with Chlamydia muridarum Protects the Oviduct from Pathology But Not from Reinfection. Infection and Immunity, 80, 2194-2203. https://doi.org/10.1128/IAI.00169-12
[11]
Hafner, L., Beagley, K. and Timms, P. (2008) Chlamydia trachomatis Infection: Host Immune Responses and Potential Vaccines. Mucosal Immunology, 1, 116-130.
https://doi.org/10.1038/mi.2007.19
[12]
Gondek, D.C., Olive, A.J., Stary, G. and Starnbach, M.N. (2012) CD4+ T Cells Are Necessary and Sufficient to Confer Protection against Chlamydia trachomatis Infection in the Murine Upper Genital Tract. The Journal of Immunology, 189, 2441-2449. https://doi.org/10.4049/jimmunol.1103032
[13]
Ito, J.I., Lyons, J.M. and Airo-Brown, L.P. (1990) Variation in Virulence among Oculogenital Serovars of Chlamydia trachomatis in Experimental Genital Tract Infection. Infection and Immunity, 58, 2021-2023.
[14]
Darville, T., Andrews, C.W., Laffoon, K.K., Shymasani, W., Kishen, L.R. and Rank, R.G. (1997) Mouse Strain-Dependent Variation in the Course and Outcome of Chlamydial Genital Tract Infection Is Associated with Differences in Host Response. Infection and Immunity, 65, 3065-3073.
[15]
Vicetti Miguel, R.D., Quispe Calla, N.E., Pavelko, S.D. and Cherpes, T.L. (2016) Intravaginal Chlamydia trachomatis Challenge Infection Elicits TH1 and TH17 Immune Responses in Mice That Promote Pathogen Clearance and Genital Tract Damage. PLoS ONE, 11, e0162445. https://doi.org/10.1371/journal.pone.0162445
[16]
Morrison, S.G., Farris, C.M., Sturdevant, G.L., Whitmire, W.M. and Morrison, R.P. (2011) Murine Chlamydia trachomatis Genital Infection Is Unaltered by Depletion of CD4+ T Cells and Diminished Adaptive Immunity. The Journal of Infectious Diseases, 203, 1120-1128. https://doi.org/10.1093/infdis/jiq176
[17]
Tuffrey, M., Falder, P., Gale, J. and Taylor-Robinson, D. (1986) Salpingitis in Mice Induced by Human Strains of Chlamydia trachomatis. The British Journal of Experimental Pathology, 67, 605-616.
[18]
Geisler, W.M., Suchland, R.J., Whittington, W.L. and Stamm, W.E. (2001) Quantitative Culture of Chlamydia trachomatis: Relationship of Inclusion-Forming Units Produced in Culture to Clinical Manifestations and Acute Inflammation in Urogenital Disease. The Journal of Infectious Diseases, 184, 1350-1354.
https://doi.org/10.1086/323998
[19]
Ramsey, K.H., DeWolfe, J.L. and Salyer, R.D. (2000) Disease Outcome Subsequent to Primary and Secondary Urogenital Infection with Murine or Human Biovars of Chlamydia trachomatis. Infection and Immunity, 68, 7186-7189.
https://doi.org/10.1128/IAI.68.12.7186-7189.2000
[20]
Parks, K.S., Dixon, P.B., Richey, C.M. and Hook 3rd, E.W. (1997) Spontaneous Clearance of Chlamydia trachomatis Infection in Untreated Patients. Sexually Transmitted Diseases, 24, 229-235.
https://doi.org/10.1097/00007435-199704000-00008
[21]
Golden, M.R., Schillinger, J.A., Markowitz, L. and St Louis, M.E. (2000) Duration of Untreated Genital Infections with Chlamydia trachomatis: A Review of the Literature. Sexually Transmitted Diseases, 27, 329-337.
https://doi.org/10.1097/00007435-200007000-00006
[22]
Joyner, J.L., Douglas Jr., J.M., Foster, M. and Judson, F.N. (2002) Persistence of Chlamydia trachomatis Infection Detected by Polymerase Chain Reaction in Untreated Patients. Sexually Transmitted Diseases, 29, 196-200.
https://doi.org/10.1097/00007435-200204000-00002
[23]
Brunham, R.C. and Rey-Ladino, J. (2005) Immunology of Chlamydia Infection: Implications for a Chlamydia trachomatis Vaccine. Nature Reviews, 5, 149-161.
https://doi.org/10.1038/nri1551
[24]
Hosenfeld, C.B., Workowski, K.A., Berman, S., Zaidi, A., Dyson, J., Mosure, D., et al. (2009) Repeat Infection with Chlamydia and Gonorrhea among Females: A Systematic Review of the Literature. Sexually Transmitted Diseases, 36, 478-489.
https://doi.org/10.1097/OLQ.0b013e3181a2a933
[25]
Pal, S., Fielder, T.J., Peterson, E.M. and de la Maza, L.M. (1994) Protection against Infertility in a BALB/c Mouse Salpingitis Model by Intranasal Immunization with the Mouse Pneumonitis Biovar of Chlamydia trachomatis. Infection and Immunity, 62, 3354-3362.
[26]
Wooters, M.A., Kaufhold, R.M., Field, J.A., Indrawati, L., Heinrichs, J.H. and Smith, J.G. (2009) A Real-Time Quantitative Polymerase Chain Reaction Assay for the Detection of Chlamydia in the Mouse Genital Tract Model. Diagnostic Microbiology and Infectious Disease, 63, 140-147.
https://doi.org/10.1016/j.diagmicrobio.2008.10.007
[27]
Farris, C.M., Morrison, S.G. and Morrison, R.P. (2010) CD4+ T Cells and Antibody Are Required for Optimal Major Outer Membrane Protein Vaccine-Induced Immunity to Chlamydia muridarum Genital Infection. Infection and Immunity, 78, 4374-4383. https://doi.org/10.1128/IAI.00622-10
[28]
Morrison, S.G., Su, H., Caldwell, H.D. and Morrison, R.P. (2000) Immunity to Murine Chlamydia trachomatis Genital Tract Reinfection Involves B Cells and CD4(+) T Cells But Not CD8(+) T Cells. Infection and Immunity, 68, 6979-6987.
https://doi.org/10.1128/IAI.68.12.6979-6987.2000
[29]
Asquith, K.L., Horvat, J.C., Kaiko, G.E., Carey, A.J., Beagley, K.W., Hansbro, P.M., et al. (2011) Interleukin-13 Promotes Susceptibility to Chlamydial Infection of the Respiratory and Genital Tracts. PLOS Pathogens, 7, e1001339.
https://doi.org/10.1371/journal.ppat.1001339
[30]
Caldwell, H.D., Kromhout, J. and Schachter, J. (1981) Purification and Partial Characterization of the Major Outer Membrane Protein of Chlamydia trachomatis. Infection and Immunity, 31, 1161-1176.
[31]
Schautteet, K., De Clercq, E. and Vanrompay, D. (2011) Chlamydia trachomatis Vaccine Research through the Years. Infectious Diseases in Obstetrics and Gynecology, 2011, Article ID: 963513. https://doi.org/10.1155/2011/963513
[32]
Pal, S., Peterson, E.M. and de la Maza, L.M. (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. Infection and Immunity, 73, 8153-8160. https://doi.org/10.1128/IAI.73.12.8153-8160.2005
[33]
Pal, S., Peterson, E.M., Rappuoli, R., Ratti, G. and de la Maza, L.M. (2006) Immunization with the Chlamydia trachomatis Major Outer Membrane Protein, Using Adjuvants Developed for Human Vaccines, Can Induce Partial Protection in a Mouse Model against a Genital Challenge. Vaccine, 24, 766-775.
https://doi.org/10.1016/j.vaccine.2005.08.074
[34]
Pal, S., Tatarenkova, O.V. and de la Maza, L.M. (2015) A Vaccine Formulated with the Major Outer Membrane Protein Can Protect C3H/HeN, a Highly Susceptible Strain of Mice, from a Chlamydia muridarum Genital Challenge. Immunology, 146, 432-443. https://doi.org/10.1111/imm.12520
[35]
Tifrea, D.F., Pal, S., Popot, J.L., Cocco, M.J. and de la Maza, L.M. (2014) Increased Immunoaccessibility of MOMP Epitopes in a Vaccine Formulated with Amphipols may Account for the Very Robust Protection Elicited against a Vaginal Challenge with Chlamydia muridarum. The Journal of Immunology, 192, 5201-5213.
https://doi.org/10.4049/jimmunol.1303392
[36]
Kari, L., Southern, T.R., Downey, C.J., Watkins, H.S., Randall, L.B., Taylor, L.D., et al. (2014) Chlamydia trachomatis Polymorphic Membrane Protein D Is a Virulence Factor Involved in Early Host-Cell Interactions. Infection and Immunity, 82, 2756-2762. https://doi.org/10.1128/IAI.01686-14
[37]
Crane, D.D., Carlson, J.H., Fischer, E.R., Bavoil, P., Hsia, R.C., Tan, C., et al. (2006) Chlamydia trachomatis Polymorphic Membrane Protein D Is a Species-Common Pan-Neutralizing Antigen. Proceedings of the National Academy of Sciences of the United States of America, 103, 1894-1899. https://doi.org/10.1073/pnas.0508983103
[38]
Paes, W., Brown, N., Brzozowski, A.M., Coler, R., Reed, S., Carter, D., et al. (2016) Recombinant Polymorphic Membrane Protein D in Combination with a Novel, Second-Generation Lipid Adjuvant Protects against Intra-Vaginal Chlamydia trachomatis Infection in Mice. Vaccine, 34, 4123-4131.
https://doi.org/10.1016/j.vaccine.2016.06.081
[39]
Morrison, S.G. and Morrison, R.P. (2005) A Predominant Role for Antibody in Acquired Immunity to Chlamydial Genital Tract Reinfection. The Journal of Immunology, 175, 7536-7542. https://doi.org/10.4049/jimmunol.175.11.7536
[40]
Farris, C.M. and Morrison, R.P. (2011) Vaccination against Chlamydia Genital Infection Utilizing the Murine C. muridarum Model. Infection and Immunity, 79, 986-996. https://doi.org/10.1128/IAI.00881-10
[41]
Nogueira, C.V., Zhang, X., Giovannone, N., Sennott, E.L. and Starnbach, M.N. (2015) Protective Immunity against Chlamydia trachomatis Can Engage Both CD4+ and CD8+ T Cells and Bridge the Respiratory and Genital Mucosae. The Journal of Immunology, 194, 2319-2329. https://doi.org/10.4049/jimmunol.1402675
[42]
Morrison, R.P. and Caldwell, H.D. (2001) Immunity to Murine Chlamydial Genital Infection. Infection and Immunity, 70, 2741-2751.
https://doi.org/10.1128/IAI.70.6.2741-2751.2002
[43]
Lyons, J.M., Ito Jr., J.I., Pena, A.S. and Morre, S.A. (2005) Differences in Growth Characteristics and Elementary Body Associated Cytotoxicity between Chlamydia trachomatis Oculogenital Serovars D and H and Chlamydia muridarum. Journal of Clinical Pathology, 58, 397-401. https://doi.org/10.1136/jcp.2004.021543
[44]
Coers, J., Gondek, D.C., Olive, A.J., Rohlfing, A., Taylor, G.A. and Starnbach, M.N. (2011) Compensatory T Cell Responses in IRG-Deficient Mice Prevent Sustained Chlamydia trachomatis Infections. PLOS Pathogens, 7, e1001346.
https://doi.org/10.1371/journal.ppat.1001346
[45]
Morrison, R.P. (2000) Differential Sensitivities of Chlamydia trachomatis Strains to Inhibitory Effects of Gamma Interferon. Infection and Immunity, 68, 6038-6040.
https://doi.org/10.1128/IAI.68.10.6038-6040.2000
[46]
Perry, L.L., Su, H., Feilzer, K., Messer, R., Hughes, S., Whitmire, W., et al. (1999) Differential Sensitivity of Distinct Chlamydia trachomatis Isolates to IFN-Gamma-Mediated Inhibition. The Journal of Immunology, 162, 3541-3548.
[47]
De Clercq, E., Kalmar, I. and Vanrompay, D. (2013) Animal Models for Studying Female Genital Tract Infection with Chlamydia trachomatis. Infection and Immunity, 81, 3060-3067. https://doi.org/10.1128/IAI.00357-13
[48]
Morre, S.A., Lyons, J.M. and Ito Jr., J.I. (2000) Murine Models of Chlamydia trachomatis Genital Tract Infection: Use of Mouse Pneumonitis Strain versus Human Strains. Infection and Immunity, 68, 7209-7211.
https://doi.org/10.1128/IAI.68.12.7209-7211.2000
[49]
Sturdevant, G.L. and Caldwell, H.D. (2004) Innate Immunity Is Sufficient for the Clearance of Chlamydia trachomatis from the Female Mouse Genital Tract. Pathogens and Disease, 72, 70-73. https://doi.org/10.1111/2049-632X.12164
[50]
Pal, S., Tifrea, D.F., Zhong, G. and de la Maza, L.M. (2018) Transcervical Inoculation with Chlamydia trachomatis Induces Infertility in HLA-DR4 Transgenic and Wild-Type Mice. Infection and Immunity, 86, e00722.
https://doi.org/10.1128/IAI.00722-17
[51]
Stary, G., Olive, A., Radovic-Moreno, A.F., Gondek, D., Alvarez, D., Basto, P.A., et al. (2015) A Mucosal Vaccine against Chlamydia trachomatis Generates Two Waves of Protective Memory T Cells. Science, 348, aaa8205.
https://doi.org/10.1126/science.aaa8205
