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, 103 (32), 12069-74

Monitoring the T Cell Response to Genital Tract Infection

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Monitoring the T Cell Response to Genital Tract Infection

Nadia R Roan et al. Proc Natl Acad Sci U S A.

Abstract

To date, it has not been possible to study antigen-specific T cell responses during primary infection of the genital tract. The low frequency of pathogen-specific T cells in a naïve mouse makes it difficult to monitor the initial events after antigen encounter. We developed a system to examine the response of pathogen-specific T cells in the genital mucosa after intrauterine infection. We identified the protective CD4(+) T cell antigen Cta1 from Chlamydia trachomatis and generated T cell receptor (TCR) transgenic (tg) mice with specificity for this protein. By transferring TCR tg T cells into naïve animals, we determined that Chlamydia-specific T cells were activated and proliferated in the lymph nodes draining the genital tract after primary intrauterine infection. Activated T cells migrated into the genital mucosa and secreted IFN-gamma. The development of Chlamydia-specific TCR tg mice provides an approach for dissecting how pathogen-specific T cells function in the genital tract.

Conflict of interest statement

Conflict of interest statement: No conflicts declared.

Figures

Fig. 1.
Fig. 1.
CD4+ T cell clone NR9.2 is Chlamydia-specific and protective. (a) CD4+ T cell clone NR9.2 was cultured with Chlamydia-infected or uninfected BMMs, and IFN-γ secretion was assessed by intracellular cytokine staining. Results are gated on live cells. (b) Naïve mice, C. trachomatis immune mice, and naïve mice that received 107 NR9.2 T cells were challenged i.v. with 107 IFU of C. trachomatis. Immune mice had recovered from infection 3 weeks before challenge. Spleen titers were determined 3 days after challenge.
Fig. 2.
Fig. 2.
NR1 TCR tg cells proliferate in response to Chlamydia infection in vivo. CFSE-labeled NR1 (Left) or OTII (Right) cells were transferred into C57BL/6 recipients. One day later, mice were infected i.v. with the indicated pathogen, and spleens were harvested 3 days later. Results were gated on live CD4+ Vα2+ cells to detect the NR1 TCR tg cells.
Fig. 3.
Fig. 3.
NR1 cells are activated and proliferate in the ILNs after intrauterine infection with C. trachomatis. CFSE-labeled NR1 cells were transferred into CD90.1 recipients, which were then mock-infected or infected in the uterus with 106 IFU of C. trachomatis serovar L2. Cells from the ILNs of infected CD90.1 recipients were harvested 5 (a) or 7 (b and c) days after infection. The levels of CD69 (a), CD62L (b), and CD44 (c) expression were measured on CD4+ NR1 cells. Results were gated on live CD90.2+ CD4+ cells to specifically detect the NR1 TCR tg cells.
Fig. 4.
Fig. 4.
NR1 cells proliferate preferentially in the ILNs after intrauterine infection with C. trachomatis. CFSE-labeled NR1 cells were transferred into CD90.1 recipients, which were then mock-infected or infected in the uterus with 106 IFU of C. trachomatis serovar L2. ILNs (Upper) and NDLNs (Lower) were harvested at the indicated times after infection, and proliferation of CD4+ NR1 cells was examined. Results were gated on live CD90.2+ CD4+ Vα2+ cells to specifically detect the NR1 TCR tg cells.
Fig. 5.
Fig. 5.
Antigen-experienced NR1 cells migrate to the genital tract tissue after intrauterine infection with Chlamydia. CFSE-labeled NR1 cells were transferred into CD90.1 recipients and then mock-infected or infected in the uterus with 106 IFU of C. trachomatis serovar L2. Seven days after infection, the genital tracts were removed from the mice and analyzed for the presence of the transferred NR1 cells. (a) The presence of CD90.2+ CD4+ NR1 cells was compared in the genital tracts of mock-infected and infected recipients. Results were gated on live cells. (b) The levels of CD62L, CD44, and CFSE on NR1 cells were examined in the genital tracts of mock-infected and infected mice. Results were gated on live CD90.2+ CD4+ cells to specifically detect the NR1 TCR tg cells. (c) NR1 cells from the genital tract were stimulated with phorbol 12-myristate 13-acetate/ionomycin and analyzed by flow cytometry to detect production of IFN-γ. Results are gated on live CD90.2+ CD4+ cells to specifically detect the NR1 TCR tg cells. Solid line indicates isotype control; filled histogram indicates IFN-γ.

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