MUCOSAL IMMUNE RESPONSES OF A RECOMBINANT CHLAMYDIA TRACHOMATIS MOMP PROTEIN

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 沙眼衣原体是世界上引起细菌性传播疾病的主要原因。我们已经开发了一个原型粘膜疫苗利用重组C。沙眼小鼠肺炎MOMP（主要外表面蛋白）亚单位蛋白。在该原型中，MOMP分子与作为粘膜佐剂的修饰的霍乱毒素（CTB）基因融合（rMOMP-CTB）。我们已经报道了用该疫苗鼻内免疫的小鼠产生强烈的粘膜和全身免疫应答。此外，用rMOMP-CTB粘膜免疫保护小鼠免受活的C.沙眼我们接下来研究了细胞因子在保护小鼠免受这种攻击感染中可能发挥的作用。用rMOMP体外再刺激从免疫小鼠获得的脾细胞，并在刺激后48小时收集上清液。使用细胞因子ELISA分析上清液中的Th 1（IL-2、IL-12 p40和IFN-γ）和Th 2（IL-4、IL-5和IL-10）细胞因子。用rMOMP免疫的小鼠引起Th 1（IFN-γ和IL-12 p40）和Th 2（IL-5）型应答。IFN-γ和IL-12 p40细胞因子的水平与来自C.沙眼衣原体免疫小鼠。然而，来自C.沙眼衣原体小鼠比rMOMP免疫小鼠产生更多的IL-10。 有趣的是，只有来自rMOMP小鼠的细胞诱导高水平的IL-5。这一发现具有重要意义，并且与这些小鼠在用活的沙眼衣原体攻击感染之前和之后的伊加的高全身和粘膜产生相关。已知鼠IL-5刺激受刺激的B细胞产生伊加。此外，这种细胞因子对于B细胞同种型转换、生长和分化为IgA分泌浆细胞至关重要。这些发现强烈地表明，rMOMP在rMOMP免疫的小鼠中比在C.沙眼小鼠

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Chlamydia trachomatis is the leading cause of bacterial sexually transmitted diseases in the world. We have developed a prototype mucosal vaccine utilizing the recombinant C. trachomatis mouse pneumonitis MOMP (major outer surface protein) subunit protein. In this prototype the MOMP molecule is genetically fused with modified cholera toxin (CTB) as a mucosal adjuvant (rMOMP-CTB). We have reported that mice immunized intranasally with this vaccine develop strong mucosal and systemic immune responses. In addition, mucosal immunization with rMOMP-CTB protected mice from a challenge infection with live C. trachomatis. We next investigated the role that cytokines may play in protecting mice from this challenge infection. Spleen cells obtained from immunized mice were restimulated in vitro with rMOMP, and supernatants were collected after 48 hr post-stimulation. Th1 (IL-2, IL-12p40 and IFN-y) and Th2 (IL-4, IL-5 and IL-10) cytokines in supernatants were analyzed using cytokine ELISA's. Mice immunized with rMOMP elicited Th1 (IFN-y and IL-12p40) and Th2 (IL-5) type responses. The levels of both IFN-y and IL-12p40 cytokines were comparable to those that were induced by cells from C. trachomatis immunized mice. However, cells from the C. trachomatis mice produced more IL-10 than those from rMOMP immunized mice. Interestingly only cells from rMOMP mice induced high levels of IL-5. This finding is of significance, and correlates with the high systemic and mucosal production of IgA in these mice before and after a challenge infection with live C trachomatis. Murine IL-5 is known to stimulate the production of IgA by stimulated B cells. Moreover, this cytokine is crucial for B-cell isotype switch, growth and differentiation into IgA-secreting plasma cells. These findings strongly suggest that rMOMP elicited strong Th1/Th2 cytokine responses that correlated with protective immunity in rMOMP immunized mice than in the C. trachomatis mice.