Molecular study of mouse viral resistance mechanisms.

我们实验室的长期目标是了解先天免疫系统如何在分子水平上识别和响应病原体入侵，特别是因为这种防御可能由基因组元件控制。 特别是，这项研究将集中在NK细胞介导的病毒免疫鼠巨细胞病毒（MCMV），因为它是由NKO连锁的Cmy 1基因座控制。 值得注意的是，NK细胞通过释放的细胞因子（例如IFN-γ）和直接的细胞介导的细胞毒性机制，是宿主对一系列病原体（包括病毒、细菌和原生动物寄生虫）的先天免疫防御的重要贡献者。 此外，NK细胞在小鼠巨细胞病毒（MCMV）感染期间直接限制病毒复制，这种能力与MCMV攻击期间的宿主存活率直接相关，并且NK细胞介导的免疫直接受Cmv 1基因座调节。 Cmv 1最近在远端NKC内定位，但未报告Cmv 1候选序列。 在初步数据中，我们报告说，一个额外的NKC编码的NK受体（Ly-49 H）可能参与MCMV抗性，但这个NKC基因是物理和遗传分离的CMv 1基因座。因此，我们将通过评估新型NKC内重组小鼠和Cmv 1阴性/阴性小鼠中的MCMV耐药性来确定MCMV耐药性是否需要一个以上的NKC基因座（特异性目的1）。 将从C57 BL/6小鼠和MCMV易感小鼠品系中选择新的Cmv 1编码序列。 将对这些序列进行结构和生物化学比较，以鉴定最佳Cmv 1候选序列。将使用转基因方法（特定目标2）确认Cmv 1候选序列。 最后，如果使用转基因方法获得暗示Ly 49 h在MCMV抗性免疫中的遗传数据。此外，将在C57 BL/6和几种MCMV易感小鼠品系中彻底表征该基因（特定目标3）。 因此，这些研究应该澄清我们对NK细胞识别病毒感染和Cmv 1调节的免疫的理解，因为这些目前还没有被理解的机制。

The long-term objective of our laboratory is to understand how the innate immune system recognizes and responds to pathogen invasion at the molecular level, especially since such defenses may be controlled by genomic elements. In particular, this study will focus on NK cell- mediated viral immunity to murine cytomegalovirus (MCMV) as it is controlled by the NKO linked Cmy1 locus. Notably NK cells are important contributors of host innate immune defenses to a range of pathogens including, viruses, bacteria, and protozoan parasites, through cytokines released (e.g. IFN-gamma) and direct cell-mediated cytotoxicity mechanisms. Moreover, NK cells directly limit viral replication in mice during murine cytomegalovirus (MCMV) infection, this capacity directly correlates with host survival during MCMV challenge, and NK cell-mediated immunity is directly regulated by the Cmv1 locus. Cmv1 was recently mapped within the distal NKC, however Cmv1 candidate sequences were not reported. In preliminary data, we report that an additional NKC encoded NK receptor (Ly- 49H) may be involved in MCMV resistance, but this NKC gene is physically and genetically separate from the CMv1 locus. Hence, we will determine whether more than one NKC locus is required MCMV resistance by assessing MCMV resistance in novel intra-NKC recombinant mice and Cmv1 minus/minus mice (Specific Aim 1). Novel Cmv1 coding sequences will be selected form C57BL/6 mice and from MCMV susceptible strains of mice. These sequences will be compared structurally and biochemically to identify a best Cmv1 candidate sequence. Confirmation of Cmv1 candidate sequences will be performed using a transgenic approach (Specific Aim 2). Finally, should genetic data be obtained implicating Ly49h in MCMV resistance immunity using a transgenic approach. Furthermore, this gene will be thoroughly characterized in C57BL/6 and several MCMV susceptible mouse strains (Specific Aim 3). Thus, these studies should clarify our understanding of NK cell recognition of viral infection and Cmv1-regulated immunity, as these are not currently understood mechanistically.