Mechanisms of NK Recognition of HCMV Infected Cells

NK识别HCMV感染细胞的机制

• 批准号: 6422026
• 负责人: WILLIAM H CARR
• 金额: $ 7.2万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6422026
• 关键词: MHC class I antigen; antibody receptor; antigen antibody reaction; blocking antibody; cellular immunity; clinical research; cytolysis; cytomegalovirus; flow cytometry; fluoresceins; host organism interaction; human tissue; immunofluorescence technique; immunogenetics; immunoregulation; molecular cloning; monoclonal antibody; mutant; natural killer cells; polymerase chain reaction; receptor binding; receptor expression; virus infection mechanism

DESCRIPTION (provided by applicant): The defense against viral pathogens by human natural killer (NK) cells is a significant area of medical research because a deficiency in NK cells results in debilitating disease or even death due to herpesvirus infections by viruses such as human cytomegalovirus (HCMV). NK cells express a diverse repertoire of receptors killer immunoglobulin-like receptors (KIR), that are specific for major histocompatibility complex (MHC) class I. Preliminary data suggest a heterogeneous response among NK cell clones in their ability to detect and lyse HCMV infected cells, which undergo virally-mediated MHC class I downregulation. The objective of this four-year study is to determine the mechanisms of human NK cell recognition of HCMV infected cells. The hypothesis is that distinct subsets of NK cells, defined by their expression of particular KIR are responsible for the recognition of HCMV infected cells through their detection of decreased MHC class I expression. This will be tested through two specific aims: 1) to determine the role of inhibitory receptors in the detection of HCMV infection and 2) to determine the role of MHC class I expression in the detection of HCMV infection. The methods used in these approaches include: flow cytometry and sequence specific primer typing of receptor expression on NK clones, in vitro cell killing assays and the use of a mutant HCMV strain, RV798 that lacks the ability to downregulate class I expression. Unlike previous studies that use unnatural, allogeneic cells, only autologous infected skin fibroblasts will be used. Dr. Carr, the principal investigator, has an extensive interest in innate immunity, through his prior clinical experience as an aquatic veterinarian in shrimp aquaculture. Dr. Carr's long term goal of conducting research in cellular immunity is advanced through both the research/ training plan and the exceptional training environment at Stanford University, the performance site. The training plan supports the completion of Dr. Carr's Ph.D. training at Stanford and his transition to a postdoctoral fellow position at the University of California at San Francisco (UCSF). This plan builds upon Dr. Carr's previous experience through the acquisition of new research skills and expertise in molecular biology, human cell culture, cellular immunology, immunogenetics and herpesvirology. Stanford offers intensive training opportunities including weekly scientific seminars, retreats, journal clubs, and conferences. Furthermore, co-sponsors at Stanford, Drs. Peter Parham and Edward Mocarski, are renowned and accomplished researchers in the fields of immunogenetics and herpesvirology, respectively. Similarly, collaborations with Drs. Lewis Lanier at UCSF and Joe Phillips at DNAX, a biotechnology company, provide complementary expertise in NK cell biology and cellular immunology on this project. An objective of the training plan is that Dr. Carr will be competitive for a RO1 on cellular, innate immunity to be submitted by the end of the four-year award period.

描述（由申请人提供）：通过以下方式防御病毒病原体： 人类自然杀伤（NK）细胞是医学研究的重要领域 因为NK细胞的缺乏会导致虚弱的疾病甚至死亡 由于疱疹病毒感染，例如人巨细胞病毒（HCMV）。 NK细胞表达多种杀伤免疫球蛋白样受体 主要组织相容性复合体（MHC）特异性受体（KIR） I类。初步数据表明，NK细胞之间的异质性反应 克隆检测和裂解HCMV感染细胞的能力， 病毒介导的MHC I类下调。这四年的目标 本研究旨在探讨人NK细胞识别HCMV的机制 被感染的细胞假设是不同的NK细胞亚群，定义为 通过表达特定的KIR，负责识别 HCMV感染的细胞通过其检测减少的MHC I类 表情这将通过两个具体目标进行测试：1）确定 抑制性受体在检测HCMV感染中的作用; 2） 确定MHC I类表达在HCMV检测中的作用 感染这些方法中使用的方法包括：流式细胞术和流式细胞仪。 体外NK克隆上受体表达的序列特异性引物分型 细胞杀伤测定和使用突变HCMV株，RV 798，其缺乏 下调I类表达的能力。不像以前的研究， 非天然的同种异体细胞，只有自体感染的皮肤成纤维细胞将被 采用卡尔博士，主要研究者，有广泛的兴趣， 先天免疫，通过他以前作为水生动物的临床经验， 养虾兽医。卡尔博士的长期目标是 通过研究/培训， 计划和特殊的培训环境在斯坦福大学， 演出现场。培训计划支持完成卡尔博士的 博士在斯坦福大学接受培训， 加州大学弗朗西斯科分校（UCSF）的一个职位。这个计划 卡尔博士以前的经验，通过收购新的 在分子生物学、人类细胞培养、 细胞免疫学、免疫遗传学和疱疹病毒学。斯坦福大学提供 强化培训机会，包括每周一次的科学研讨会， 务虚会、期刊俱乐部和会议。此外，共同赞助者在 斯坦福大学，彼得帕勒姆和爱德华莫卡斯基博士，是著名的和有成就的 免疫遗传学和疱疹病毒学领域的研究人员。 同样，与加州大学旧金山分校的刘易斯拉尼尔博士和加州大学旧金山分校的乔菲利普斯博士的合作也是如此。 DNAX是一家生物技术公司，提供NK细胞方面的补充专业知识 生物学和细胞免疫学。培训的目标 计划是，卡尔博士将有竞争力的RO 1细胞，先天 豁免权将在四年授标期结束前提交。