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 central importance of virus-specific CD8+ and CD4+ T lymphocytes in containing HIV-1 replication has been appreciated. It will be important to characterize these lymphocytes in greater depth to determine how they contribute to containing HIV-1 replication. However, our ability to study these lymphocyte populations has been limited by the technologies available to carry out such analyses. Tetrameric MHC class I and II/peptide complexes that label AIDS virus-specific subpopulations of T cells has provided an important tool to visualize those cell populations. By providing a reproducible and precise quantitative assay for CTL and CD4+ T cell responses, the tetramer assay has dramatically increased the level of sophistication that can be brought to bear on the study of antigen-specific T lymphoctes. However, the application of the tetramer technology to these responses has been focus on the analysis of single cell suspension by flow cytometry. Understanding the spatial relationship and distribution of these cells in different tissues should increase our ability to evaluate their role in disease pathogenesis. In the studies described in this application, we will evaluate other dyes to improve the intensity and specificity of the in situ MHC class I tetramer staining and develop MHC class II tetramer for the same application.
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