Characterization of Human T Cells Against Chlamydia

描述(申请人提供)：沙眼衣原体(Ct)最多 世界范围内细菌性传播疾病的常见原因。在以前的 工作中，我们确定了主要外膜(MOMP)中的表位 在人类白细胞抗原II类分子等背景下被CD4+T细胞识别 是由CD8+T细胞在人类白细胞抗原I类分子的背景下识别的。 最近，我们使用HLAA2四聚体来鉴定MOMP特异性CD8+T细胞 在CT感染者的血液中发现他们杀死了CT感染者 上皮细胞。我们将把这些研究扩展到其他使用人类白细胞抗原的个体 用其他同种异型制成的第I类四聚体。识别和刻画 从血液中分离出MOMP特异性的CD4+T细胞，我们将创建人类白细胞抗原II类 使用可被CD4+T细胞识别的MOMP表位的四聚体。我们会 CD8和CD4MOMP特异性T细胞的细胞表面特征 参与归巢的蛋白质(例如整合素和趋化因子受体)， 功能(如细胞因子、穿孔素)和分化状态(如HLA-DR、 CD62L)，用抗CD62L的单抗进行多色流式细胞术 细胞表面和细胞内的蛋白质。不同来源的淋巴细胞的比较 血液与感染部位的对比将提供有关代表性的信息 血液样本是感染部位的细胞。最后，我们有能力 MOMP特异性CTL抑制感染生殖器中感染性CT的增加 TRAIL细胞及其CTL表位突变对这种限制的影响 将会被确定。检测和描述人类特征的能力 抗衣原体T细胞与四聚体和确定的人类白细胞抗原-1细胞试剂 表位特异性将有助于疫苗的开发，特别是 因为大多数表位位于MOMP的保守区。

DESCRIPTION (provided by the applicant): Chlamydia trachomatis (Ct) is the most common cause of bacterial sexually transmitted disease worldwide. In previous work, we identified epitopes in the major outer membrane (MOMP) that are recognized by CD4+ T cells in the context of HLA class II molecules and others that are recognized by CD8+ T cells in the context of HLA class I molecules. Most recently, we used HLA A2 tetramers to identify MOMP-specific CD8+ T cells in blood of Ct-infected individuals and found that they killed Ct infected epithelial cells. We will extend these studies to other individuals using HLA class I tetramers made with other allotypes. To identify and characterize MOMP-specific CD4+ T cells isolated from blood, we will create HLA class II tetramers using MOMP epitopes that are recognized by CD4+ T cells. We will characterize both CD8 and CD4 MOMP-specific T cells with regard to cell surface proteins that are involved in homing (e.g. integrins and chemokine receptors), function (e.g. cytokines, perforin) and state of differentiation (e.g. HLA-DR, CD62L) by using multi-color flow cytometry with monoclonal antibodies against cell surface and intracellular proteins. Comparisons of lymphocytes from the blood vs. sites of infection will provide information on how representative blood-derived samples are of cells at infected sites. Lastly, the ability of MOMP-specific CTL to limit the increase of infectious Ct in infected genital tract cells and the effects of mutations in CTL epitopes on such limitation will be determined. The ability to detect and characterize human anti-Chlamydial T cells with tetramers and cellular reagents of defined HLA- and epitope specificity would be useful for vaccine development, especially since most of the epitopes are in conserved regions of MOMP.