Effective immune responses require the development of T lymphocytes specific for antigens presented by self major histocompatibility complex (MHC) class I and class II molecules. Many basic and clinical studies require the detection and enumeration of antigen- specific T lymphocytes, but until recently their detection was inaccurate and cumbersome, requiring extensive tissue culture procedures. New reagents have been developed, comprised of four identical MHC molecules, each presenting a defined antigenic peptide. The reagent is fluorescently labeled to enable the direct detection of T cells that bind the reagent by their specific antigen receptor molecules. Previous research has established the approach for class I MHC molecules that bind to cytotoxic T cells, but reagents based upon class II MHC molecules, which would detect helper T lymphocytes, are less well-developed. This contract will focus on the development and utilization of class II MHC-peptide tetramers to detect and study antigen specific T cells. Studies will include: a) testing various forms of class II MHC-peptide complexes to optimize the reagents, b) demonstrate the reagents~ ability to detect human T cells in clinical samples and c) combine detection with functional T cell assays to increase the information obtained. Because helper T cells are central to virtually all immune responses, this technology is applicable to studies in many areas, including basic immune mechanisms, infectious diseases, vaccination, autoimmunity, transplant rejection, and tumor therapy.
| Stone, Jennifer D; Demkowicz Jr, Walter E; Stern, Lawrence J (2005) HLA-restricted epitope identification and detection of functional T cell responses by using MHC-peptide and costimulatory microarrays. Proc Natl Acad Sci U S A 102:3744-9 |
| Cameron, Thomas O; Cohen, George B; Islam, Sabina A et al. (2002) Examination of the highly diverse CD4(+) T-cell repertoire directed against an influenza peptide: a step towards TCR proteomics. Immunogenetics 54:611-20 |
| Cameron, T O; Cochran, J R; Yassine-Diab, B et al. (2001) Cutting edge: detection of antigen-specific CD4+ T cells by HLA-DR1 oligomers is dependent on the T cell activation state. J Immunol 166:741-5 |
| Cochran, J R; Aivazian, D; Cameron, T O et al. (2001) Receptor clustering and transmembrane signaling in T cells. Trends Biochem Sci 26:304-10 |
| Cochran, J R; Stern, L J (2000) A diverse set of oligomeric class II MHC-peptide complexes for probing T-cell receptor interactions. Chem Biol 7:683-96 |
