HERPES SIMPLEX VIRUS GE AND GI PROTEIN SPECIFIC T-CELLS

单纯疱疹病毒 GE 和胃肠道蛋白特异性 T 细胞

• 批准号: 3078881
• 负责人: PAMELA DIAZ
• 金额: $ 7.73万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-08-01 至 1994-07-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3078881
• 关键词: CD4 molecule; CD8 molecule; Herpes simplex disease; MHC class I antigen; MHC class II antigen; cell mediated cytotoxicity; cytotoxic T lymphocyte; disease /disorder model; flow cytometry; glycoprotein biosynthesis; glycoproteins; herpes simplex virus 1; laboratory mouse; lymph nodes; molecular cloning; monoclonal antibody; protein sequence; surface antigens; tissue /cell culture; transfection; virus cytopathogenic effect; virus infection mechanism; virus protein

The candidate is a pediatrician who has completed her clinical training in pediatric infectious diseases. Her goal is to pursue an academic career in this subspeciality. Her preliminary research experience has allowed her to identify viral immunology as her general area of research interest. She seeks a Clinical Investigator Award for three years of support in order to acquire the necessary theoretical background and laboratory experience to become a competent independent investigator. She has designed a didactic program and a proposal for supervised laboratory research under the sponsorship of Dr. Bernard Roizman, Molecular Genetics and Cell Biology, and Dr. Jeff Bluestone, Pathology. The research proposal focuses upon the T-lymphocyte immunity to herpes simplex virus in the mouse model. Herpes simplex virus causes serious disease in previously healthy and immunocompromised patients. There is evidence that T-lymphocyte immunity plays a major role in the immunity to herpes viruses in humans during the recovery from acute infection. The role that specific viral proteins play in eliciting cytotoxic T-cells will be studied. As components of the viral envelope, HSV glycoproteins are likely to be important in inducing immunity. The purpose of this proposal is to examine the interaction of cytotoxic T-lymphocytes with HSV-1 glycoprotein E (gE), glycoprotein I (gI), and the glycoprotein E/glycoprotein I (gE/gI) complex in the murine model of HSV-1 infection and to correlate the host response to these proteins with disease pathogenesis. The proposal consists of two major phases. In the first phase, gE and gI will be cloned and then transfected into a major histocompatibility complex (MHC) matched murine fibroblast cell line which expresses both class I and II MHC proteins. These constructed cell lines will be used in the second phase of the project as stimulating antigen for the cloning of HSV specific T-cells and targets in the cytotoxicity assays. CBA mice will be infected by footpad injection with HSV-1 and after six days their draining popliteal lymph nodes will be isolated and cultured in vitro. Cultures exhibiting cytolytic capabilities will be characterized as to their phenotypic markers (i.e. CD3, CD4, CD8, MHC-1, MHC-II). T-cell surface antigens will be studied by fluorescence activated cell sorting analysis. Adoptive transfer experiments with protein specific cloned T-cells will provide a measure of the protective efficacy of gE, gI, or gE/gI CTL given during acute infection. These studies will help to delineate the effect that gE and gI have upon the immune system and whether their inclusion in any HSV vaccine would be beneficial or detrimental to the host.

该候选人是一名儿科医生，她在 儿科传染病 她的目标是追求学术生涯， 这个subspeciality。 她的初步研究经验使她能够 确定病毒免疫学为她研究兴趣的一般领域。 她 寻求临床研究者奖三年的支持，以 获得必要的理论背景和实验室经验， 成为一名合格的独立调查员。 她设计了一个教学 计划和监督下的实验室研究的建议 分子遗传学和细胞生物学伯纳德·罗伊兹曼博士的赞助， 和病理科的杰夫·布鲁斯通医生 研究计划的重点是 单纯疱疹病毒感染小鼠模型的t淋巴细胞免疫。 疱疹 单纯型病毒导致严重的疾病， 免疫功能低下的患者。 有证据表明T淋巴细胞免疫 在人类疱疹病毒免疫中起主要作用， 从急性感染中恢复过来。 特定的病毒蛋白质所起的作用 将研究诱导细胞毒性T细胞的作用。 作为病毒的成分 包膜，HSV糖蛋白可能是重要的诱导 免疫力 本提案的目的是审查以下方面的相互作用： 具有HSV-1糖蛋白E（gE）、糖蛋白I的细胞毒性T淋巴细胞 (gI)和小鼠中的糖蛋白E/糖蛋白I（gE/gI）复合物 HSV-1感染的模型，并将宿主反应与这些 蛋白质与疾病的发病机制。 该提案包括两个主要部分 阶段。 在第一阶段，gE和gI将被克隆，然后转染 主要组织相容性复合体（MHC）匹配的小鼠成纤维细胞 表达I类和II类MHC蛋白的细胞系。 这些 构建的细胞系将用于项目的第二阶段， 刺激抗原用于克隆单纯疱疹病毒特异性T细胞和靶点 细胞毒性试验。 CBA小鼠将通过足垫注射感染 HSV-1感染，六天后，他们的引流腘淋巴结将被 分离培养。 显示细胞溶解能力的培养物 将根据其表型标志物（即CD3，CD4，CD8， MHC-I、MHC-II）。 T细胞表面抗原将通过荧光研究 活化细胞分选分析。 连续转移实验， 蛋白质特异性克隆T细胞将提供一种保护性措施， 急性感染期间给予gE、gI或gE/gI CTL的疗效。 这些 研究将有助于阐明gE和gI对 免疫系统，以及它们是否包含在任何HSV疫苗中， 对宿主有益或有害。