Mechanisms of NK Recognition of HCMV Infected Cells

描述(申请人提供)：对病毒病原体的防御 人类自然杀伤(NK)细胞是医学研究的重要领域 因为NK细胞缺乏会导致衰弱的疾病甚至死亡 由于疱疹病毒感染人类巨细胞病毒(HCMV)等病毒。 NK细胞表达不同的受体杀伤免疫球蛋白样谱 主要组织相容性复合体(MHC)特异性受体 第I类初步数据显示NK细胞之间存在异质性反应 克隆人检测和裂解人巨细胞病毒感染细胞的能力 病毒介导的MHC I类下调。这四年的目标是 研究目的是确定人巨细胞病毒对人NK细胞的识别机制 被感染的细胞。假设是不同的NK细胞亚群，定义为 通过他们对特定KIR的表达，负责识别 人巨细胞病毒通过检测MHC-I类分子的减少感染细胞 表情。这将通过两个具体目标进行测试：1)确定 抑制受体在检测巨细胞病毒感染中的作用和2) 确定MHC-I类表达在检测巨细胞病毒中的作用 感染。这些方法中使用的方法包括：流式细胞术和 自然杀伤细胞克隆受体表达的序列特异性引物分型 细胞杀伤试验和使用突变的巨细胞病毒株RV798，它缺乏 下调I类基因表达的能力。与以前的研究不同，使用 非自然的，同种异体细胞，只有自体感染的皮肤成纤维细胞才会 使用。卡尔博士，首席研究员，对 先天免疫力，通过他以前作为水生生物的临床经验 对虾水产养殖业兽医。卡尔博士的长期目标是进行 细胞免疫的研究是通过研究/培训来推进的 计划和斯坦福大学卓越的培训环境， 表演现场。该培训计划支持完成卡尔博士的 斯坦福大学的博士培训和他向博士后的转变 加州大学旧金山分校(UCSF)的职位。这个计划 在卡尔博士先前经验的基础上，通过收购新的 分子生物学、人类细胞培养等方面的研究技能和专业知识 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，包括每周科学研讨会， 静修、日记俱乐部和会议。此外，联合赞助商在 斯坦福大学，彼得·帕拉姆博士和爱德华·莫卡斯基博士，是著名的和有成就的 分别是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·拉尼尔博士和乔·菲利普斯博士在 DNAX是一家生物技术公司，提供NK细胞方面的互补专业知识 生物学和细胞免疫学在这个项目上。培训的一个目标 计划是卡尔博士将在细胞上竞争RO1，与生俱来 豁免权须在四年授权期结束前提交。

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.