MUCOSAL IMMUNITY TO CHLAMYDIAL INFECTION

该项目的目标是直接确定衣原体是否 特异性分泌型免疫球蛋白A(SIgA)在保护机体免受 小鼠生殖道衣原体感染的消退。按顺序 为了解决这个问题，我们将生产出能分泌 衣原体特异性聚合型IgA(PIgA)单抗 表面成分，以及2)确定其防护能力 衣原体生殖道感染的小鼠模型。单抗将会是 通过皮下植入分泌的mAb给幼年动物注射 同基因小鼠背部杂交瘤(背包肿瘤系统)。这 这种方法之所以成为可能，是因为系统交付的PIGA 特异性地与分泌片络合并转运到粘膜 表面为SIgA，而单体Ig A(MIGA)和Ig G不是。到目前为止， 已经产生了三种产生PIgA特异性的小鼠杂交瘤 沙眼衣原体MoPn.这些杂交瘤是由 肠系膜淋巴来源淋巴细胞的融合实验 免疫球蛋白A表面阳性细胞丰富的群体。使用PIGA 和分泌免疫球蛋白的杂交瘤，我们目前正在进行一项试验 在我们实验室进行背包肿瘤系统的实验研究。 初步数据显示，肿瘤发展的采集率为 -gt；80%，这种肿瘤产生高水平的mAb，而该肿瘤 不同肿瘤细胞系的生长速度差异很大。有了这个 信息，我们现在可以评估抗衣原体SIgA在 对生殖道粘膜衣原体定植的保护作用 以及它在解决已确定的生殖道感染方面的作用。 相同的单抗集合将允许未来的研究在 体外中和衣原体感染性AS的体内保护作用 黏膜特异性IgA抗体依赖性的研究 细胞毒性作为保护性免疫的一种潜在机制。

The goal of this project is to directly determine whether Chlamydia specific secretory IgA (sIgA) plays a role in protection from, or resolution of, chlamydial infection of the mouse genital tract. In order to address this question we will 1) produce murine hybridomas that secrete polymeric IgA (pIgA) monoclonal antibodies (mAbs) specific to chlamydial surface components, and 2) determine their ability to protect against chlamydial genital tract infection in a murine model. mAbs will be administered to naive animals by subcutaneously implanting a mAb secreting hybridoma in the backs of syngeneic mice (backpack tumor system). This approach is made possible by the fact that systemically delivered pIgA is specifically complexed with secretory piece and transported to mucosal surfaces as sIgA, whereas monomeric IgA (mIgA) and IgG are not. To date, three murine hybridomas have been generated that produce pIgA specific for the surface of Chlamydia trachomatis MoPn. These hybridomas resulted from fusion experiments using mesenteric lymph node derived lymphocyte populations that were enriched for IgA surface positive cells. Using pIgA and IgG secreting hybridomas, we are currently conducting a pilot experiment to characterize the backpack tumor system in our laboratory. Preliminary data indicate that the take rate for tumor development is >80%, that high levels of mAb are produced by such tumors, and that tumor growth rates vary significantly between tumor cell lines. With this information, we can now evaluate the role of anti-chlamydial sIgA in protection against chlamydial colonization of the genital tract mucosae as well as its role in resolving established infections of the genital tract. The same collection of mAbs will allow for future studies that compare in vivo protection with in vitro neutralization of chlamydial infectivity, as well as, investigations of mucosae specific IgA antibody dependent cellular cytotoxicity as a potential mechanism of protective immunity.