DEVELOPMENT OF CORE SERVICE FOR SIV CELLULAR IMMUNE ASSAYS

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。细胞免疫学核心是由Marcelo黑田博士领导的免疫学部门的一部分。该中心由一名医学研究专家组成，根据要求为内部和外部研究人员的特定研究项目提供免疫学服务。目前的服务包括在标准化程序下进行样品制备以进行最佳测定分析、ELISPOT测定的规划和执行、数据处理和呈现、细胞内细胞因子染色以及制备重组牛痘病毒原液以用作这些测定中的抗原刺激剂。 我们将继续使用MabTech ELISPOT系统以及U-Cytech系统，具体取决于特定的项目和合作者。在大多数情况下，我们继续使用Zellnet咨询服务来记录和分析ELISPOT结果。我们为6名内部研究者和2名外部合作者提供了ELISPOT服务，涉及5个不同的项目，分析了112个动物时间点对多种重组抗原的反应。我们已经制备了2种重组牛痘病毒的高滴度储备液，用于一个特定项目。我们已经为4个不同的项目准备了来自61只动物的PBMC和/或淋巴结活检的冷冻保存的单核细胞样品。细胞免疫学核心还为研究人员免费提供MHC分型服务。在这一年中，超过50只恒河猴被分配到四个正在进行的项目中，在TNPRC进行了测试，以确定是否存在特定的恒河猴MHC-I和II基因。Core已实施PCR检测，可准确确定Mamu A01、Mamu A02和Mamu-DRw 201分子的存在。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The Cellular Immunology Core is a part of the Immunology Division headed by Dr. Marcelo Kuroda. The core is staffed by one Medical Research Specialist, who provides immunology services to specific research projects of in-house and outside investigators as requested. Current services include sample preparation under standardized procedures for optimal assay analysis, planning and performance of ELISPOT assays, data processing and presentation, intracellular cytokine staining, and preparation of recombinant vaccinia viral stocks for use as antigenic stimulants in these assays. We are continuing to use the MabTech ELISPOT system as well as the U-Cytech system depending on the particular project and collaborators. For the most part we have continued to use the Zellnet Consulting service for recording and analyzing the ELISPOT results. We have provided ELISPOT service to 6 in-house investigators and 2 outside collaborators on 5 distinct projects resulting in analysis of 112 animal time-points for responses to up to multiple recombinant antigens. We have prepared high-titered stocks of 2 recombinant vaccinia viruses for use in one particular project. We have prepared cryopreserved mononuclear cell samples of PBMC and/or lymph node biopsies from 61 animals for 4 distinct projects. The Cellular Immunology Core also provides MHC typing service free of cost to investigators. During this year, more than fifty Rhesus monkeys assigned to four ongoing projects at the TNPRC were tested for the presence of a particular Rhesus monkey MHC-I and II genes. The Core has implemented PCR test that accurately determined the presence Mamu A01, Mamu A02 and Mamu-DRw201 molecules.