We propose to study the repertoire of the immune response localized to tissues at sites of inflammation. These studies shall focus on the host response to allografts as demonstrated by the cells which can be propagated directly from biopsy tissue of grafts undergong episodes of cellular rejection. The ability of Interleukin 2 (IL2) to provide a mitogenic stimulus to """"""""activated"""""""" T lymphocytes permits the outgrowth of host cells, from allograft tissue, which have been stimulated by the gtaft antigens. We shall determine the surface phenotype of tissue infiltrating cells in comparison to blood lymphocytes from the same patients and test the function and specificity of these cells in mixed lymphocyte and cell mediate lympholysis assays. Subsets of tissue infiltrating lymphocytes will be isolated by sorting on a FACS 440 by two color parameters in order to select functionally distinct cell types and determine their interactive roles in allograft recognition and destruction (or tolerance). Clones of tissue-infiltrating cells will be isolated to determine on a single cell level the fine specificity of alloreactions in transplant recipients, as well as interactional events between individual clones. Finally, we propose to use Southern blotting techniques in order to assess diversity, or clonal dominace of the immune response in situ. The cDNA probes now available to assist in the dissection of the T lymphocyte repertoire offer the capacity to correlate dominant patterns of genetic rearrangement with specificity and function of the cells which show restricted, or identical patterns of gene rearrangement for the T lymphocyte receptor. While the studies will focus on the allograft, preliminary evidence is presented which suggests the applicability of the techniques described for dissecting the immune response to other inflammatory stimuli, including auto-immune disorders and tumors.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA044324-03
Application #
3186898
Study Section
Surgery, Anesthesiology and Trauma Study Section (SAT)
Project Start
1986-08-01
Project End
1990-03-31
Budget Start
1989-04-01
Budget End
1990-03-31
Support Year
3
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02199
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Pandolfi, F; Oliva, A; Sacco, G et al. (1993) Fibroblast-derived factors preserve viability in vitro of mononuclear cells isolated from subjects with HIV-1 infection. AIDS 7:323-9
Pandolfi, F; Trentin, L; Boyle, L A et al. (1992) Expression of cell adhesion molecules in human melanoma cell lines and their role in cytotoxicity mediated by tumor-infiltrating lymphocytes. Cancer 69:1165-73
Bennett, W T; Pandolfi, F; Grove, B H et al. (1992) Dominant rearrangements among human tumor-infiltrating lymphocytes. Analysis of T-cells derived from 32 patients with melanoma, lung, and renal cell carcinoma. Cancer 69:2379-84
Pandolfi, F; Trentin, L; San Martin, J E et al. (1992) T cell heterogeneity in patients with common variable immunodeficiency as assessed by abnormalities of T cell subpopulations and T cell receptor gene analysis. Clin Exp Immunol 89:198-203
Frisman, D M; Fallon, J T; Hurwitz, A A et al. (1991) Cytotoxic activity of graft-infiltrating lymphocytes correlates with cellular rejection in cardiac transplant patients. Hum Immunol 32:241-5
Pandolfi, F; Boyle, L A; Trentin, L et al. (1991) Expression of HLA-A2 antigen in human melanoma cell lines and its role in T-cell recognition. Cancer Res 51:3164-70
Chatila, M K; Pandolfi, F; Stamenkovich, I et al. (1990) Clonal dominance among synovial tissue-infiltrating lymphocytes in arthritis. Hum Immunol 28:252-7
Frisman, D M; Hurwitz, A A; Bennett, W T et al. (1990) Clonal analysis of graft-infiltrating lymphocytes from renal and cardiac biopsies. Dominant rearrangements of TcR beta genes and persistence of dominant rearrangements in serial biopsies. Hum Immunol 28:208-15
Scott, S; Pandolfi, F; Kurnick, J T (1990) Fibroblasts mediate T cell survival: a proposed mechanism for retention of primed T cells. J Exp Med 172:1873-6

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