CHLAMYDIA TRACHOMATIS VACCINE STUDIES IN NONHUMAN PRIMATES

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们利用沙眼衣原体重组MOMP(主要外表面蛋白)亚单位蛋白研制了一种黏膜疫苗原型。在这个原型中，MOMP分子与修饰的霍乱毒素(CTB)基因融合，作为粘膜佐剂(rMOMP-CTB)。我们的研究表明，小鼠鼻内免疫(I.N.)有了这种疫苗，他们产生了粘膜和系统免疫反应，保护他们免受活沙眼衣原体的攻击感染。本项目的目的是研究rMOMP-CTB疫苗在灵长类动物中的安全性、免疫原性和有效性。研究对象为5只性成熟雌性恒河猴，随机分为3组：未感染对照组(n=1)、疫苗接种+攻毒组(n=2)和单纯攻毒组(n=2)。三个200微克剂量的疫苗被肌肉注射。每隔4周向每只接种疫苗的动物发放一次疫苗。所有动物每隔2周至12周采集血液、尿液和阴道冲洗液，用于安全性和免疫原性研究。安全性通过体格检查和血、尿的临床实验室分析进行评估。通过体外rMOMP诱导的外周血单个核细胞(PBMC)和扁桃体细胞的增殖反应和细胞因子召回反应来评估免疫原性。采用流式细胞术检测T/B细胞和单核细胞在PBMC中的分布和活化情况。用血清和阴道冲洗液检测rMOMP特异性抗体Ig G、Ig A和Ig M。在第12周和第13周对接种组和仅接种沙眼衣原体的2只动物阴道内注射两剂沙眼衣原体活疫苗(4.75×105)来评估疫苗的效力。在第一次接种后第12周和第15周进行阴道镜检查。在第一次激发剂量后每周收集阴道拭子以计数衣原体。所有动物在18周时被处死。收集组织(阴道、宫颈、输卵管和淋巴结)进行组织病理学和共聚焦显微镜研究。目前对样品进行了存档，以供处理和分析。

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. We have developed a prototype mucosal vaccine utilizing Chlamydia trachomatis recombinant 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). Our studies demonstrated that mice immunized intranasally (i.n.) with this vaccine developed mucosal and systemic immune responses that protected them from a challenge infection with live C. trachomatis. The goal of this project is to investigate the safety, immunogenicity and efficacy of the rMOMP-CTB vaccine in primates. The study consisted of 5-sexually matured female rhesus macaques divided into 3 groups: uninfected control (n=1), vaccinated and challenged (n=2) and challenged only (n=2). Three 200 microgram-doses of the vaccine were given i.n. to each of the vaccinated animals at 4-weeks intervals. Blood, urine and vaginal washes were obtained from all animals every 2 weeks up to 12 weeks for safety and immunogenicity studies. Safety was assessed by physical examination and clinical laboratory analyses of blood and urine. Immunogenicity was assessed by in vitro rMOMP-induced proliferative and cytokine recall responses in peripheral blood mononuclear cells (PBMCs) and tonsil cells. Flow cytometry was conducted to determine the distribution/activation of T/B-cells and monocytes in PBMCs. Sera and vaginal washes were used to determine rMOMP-specific IgG, IgA and IgM antibodies. Efficacy of the vaccine was assessed by administering two intravaginal challenge doses of live C. trachomatis (4.75 x 105) to the vaccinated and 2 challenged only animals at weeks 12 and 13. Colposcopies were conducted at weeks 12 and 15 after the 1st vaccination. Vaginal swabs were collected weekly after the first challenge dose to enumerate Chlamydia. All animals were sacrificed at 18 weeks. Tissues (vagina, cervix uterus, fallopian tubes and lymph nodes) were collected for histopathology and confocal microscopy studies. Samples are currently archived for processing and analyses.