Understanding the cellular populations and mechanisms in charge of overcoming immune compartmentalization is usually valuable for designing vaccination strategies targeting distal mucosae. T cell-dependent mucosal immunity (2). There are important distinctions between different mucosal tissues. For example, the lower respiratory and upper genital tracts are relatively sterile and intolerant of flora compared to the gastrointestinal tract. Another example is the unique lympho-epithelial structure of the intestinal Peyers patches, in contrast to the genital mucosa that lacks organized lymphoid elements. T cell migration among mucosal surfaces is also tightly regulated by the conversation of adhesion molecules and chemokine receptors that are differentially expressed on T cells and their target tissues (3, 4). For instance, skin-homing T cells express ligands for E- and P-selectins, as well as the chemokine receptors, CCR4 and CCR10 (5C7), while gut-homing effector and memory cells express the 47 integrin and HG-14-10-04 CCR9 chemokine receptor (8, 9). Despite these differences, the presence of shared immune elements between mucosal sites is also well acknowledged. For instance, other than well-described skin-homing properties, the E- and P-selectins are also involved in the migration of activated T cells to the peritoneal cavity during inflammation (6). Furthermore, the ability to use remote-site immunization to generate protective immunity at a distinct tissue also suggests that there are aspects of the immune system shared by numerous mucosal surfaces (10C12). Intranasal immunization with or HIV antigens has been shown to confer some protection in the genital tract and the protection is usually correlated with mucosal antibody responses and sometimes heightened cell-mediated responses (10, 12, 13). However, it is not obvious which of these elevated responses is usually responsible or sufficient for cross-mucosal protection. Given its ability to infect several mucosal sites, offers a unique opportunity to explore how tissue-specific HG-14-10-04 immunity might be overcome. is responsible for significant morbidity worldwide. Contamination of the ocular epithelium causes blinding trachoma and contamination of the genital mucosa can result in ectopic pregnancy and infertility (14C18). Moreover, if contamination of pregnant women is not detected, perinatal transmission of to the lungs of the newborn can ultimately result in pneumonia (19). Using murine contamination models, researchers have shown that although antibodies can provide limited protection against species (20, 21), the host response to contamination is primarily dependent on IFN (22C26). Both CD4+ and CD8+ T cells are stimulated during contamination and secrete IFN. However, elimination of CD8+ T cell response does not appear to compromise protection against genital contamination (20, 27, 28). In contrast, CD4+ T cells are both necessary and sufficient to confer protection against subsequent contamination (22, 29). The signals that govern CD4+ T cell trafficking to the genital mucosa have not been completely elucidated but it is known that efficient migration of antigen Cta1133C152 have been explained previously (25). CXCR3?/?CCR5?/? mice were generated by crossing CXCR3?/? and CCR5?/? mice. Mice were managed within the Harvard Medical School Center for Animal Resources and Comparative Medicine. All experiments in this statement were approved by Harvards Institutional Animal Care and Use Committee. Growth, isolation, and detection of bacteria serovar L2 (434/Bu) was propagated within McCoy cell monolayers as previously explained (30, 31). Aliquots of purified elementary bodies were stored at ?80 C in medium containing 250 mM sucrose, 10 mM sodium phosphate, and 5 mM L-glutamic acid (SPG). Contamination of mice and preparation of tissue For intranasal inoculation, mice were sedated with 5% isoflurane HG-14-10-04 (Vedco Inc, St. Joseph, MO) in oxygen and inoculated with 40 L SPG made up of 105 IFU of was deposited using the NSET pipet tip (ParaTechs, Lexington, KY). Uteri were minced with scalpels and enzymatically dissociated in HBSS/Ca2+/Mg2+ made up of 1 mg/ml type XI collagenase and 50 Kunitz/ml DNase for 30 HG-14-10-04 minutes at 37 C, washed in PBS made up of 5 mM EDTA, and ground between microscope slides before filtration through a 70-m mesh (32). To Rabbit polyclonal to GnT V determine levels in systemic organs, peripheral blood was collected in 10% sodium.