Virology and Technology Core

病毒学和技术核心的目的有两个方面：1）提供设施和技术 有助于在小鼠中进行病毒感染和测定，和2）促进使用新的 我们已经开发了每个项目的检测试剂盒，用于检测针对同种抗原的免疫反应。 生物安全2/3级（BSL 2/3）生物防护套间内的一个房间将被分配用于与 病毒，用于随后的监视和监测，以及用于小鼠放血和收获 组织中对于病毒和免疫检测，我有一个BSL 2实验室区域， 组织培养室，配有培养箱和层流罩，用于组织培养工作。这一核心将 负责病毒的生长、制备、纯化和测定，维持肽的储备 表位，以及用于制备抗原特异性MHC-多聚体试剂（MHC-Ig-二聚体和MHC四聚体）。 核心将提供试剂和检测培训，以量化和表征病毒特异性 包括使用MHC-多聚体、T细胞增殖测定、细胞内细胞因子测定和细胞内免疫测定， 抗病毒CTL测定。所使用的新技术包括体内细胞毒性测定，以测量NK细胞和 T细胞对同种异体组织的活性，以及细胞内细胞因子测定来检查幼稚的、 效应/记忆和耐受化同种异体反应性T细胞。核心小组将提供服务， 中心还将负责培训人员，

The purpose ofthe Virology and Technology Core is two fold: 1) to provide facilities and technical assistance for the conduction of virus infections and assays in mice, and 2) to facilitate the use of novel assays that we have developed by each ofthe Projects to examine immune responses against alloantigens. One room within a Biosafety level two/three (BSL2/3) Biocontainment Suite will be assigned for work with viruses, for their subsequent surveillance and monitoring, and for the bleeding of mice and harvesting of tissues. For viral and immunological assays, I have a BSL2 laboratory area that has a separate attached tissue culture room equipped with incubators and laminar flow hoods for tissue culture work. This Core will be responsible for the growth, preparation, purification, and assay of viruses, maintaining stocks of peptide epitopes, and for the preparation of antigen-specific MHC-multimer reagents (MHC-lg-dimers and MHCtetramers). The Core will provide reagents and training for assays to quantify and characterize virus-specific T cells including the use of MHC-multimers, T cell proliferation assays, intracellular cytokine assays, and antiviral CTL assays. The new technology to be used include in vivo cytotoxicity assays to measure NK and T cell activity against allogeneic tissues, and intracellular cytokine assays to examine naive, effector/memory, and tolerized alloreactive T cells. The services to be provided by the Core will be available to all Project Laboratories, and the Core will also be responsible for training personnel to peri'orm assays