DEVELOPMENT OF CORE SERVICE FOR SIV CELLULAR IMMUNE ASSAYS

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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.