Isolation, Functional Properties and Clonal Analysis of Tumor-Infiltrating Lymphocytes from Human Brain Tumors
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Despite the numerous reports of decreased or defective systemic cell mediated immunity in glioma patients (1–7) there is evidence of a local immune response to the tumor. Thus mononuclear cell (MNC) infiltrates have been demonstrated within the parenchyma of human glial tumors in 30–60 % of cases reported in the literature (8, 9). Other studies have correlated the intensity of MNC infiltrates with tumor histology and survival in glioma patients (10–12). Von Hanwehr and co-workers (9) found that T lymphocytes constituted the major fraction of the infiltrates in gliomas. However the nature and functional capabilities of these cells and their potential cytolytic role in preventing tumor growth remain poorly understood. To resolve these questions it is necessary to isolate the tumor infiltrating lymphocytes (TIL) from the tumor tissue. To this end TIL from 7 human brain tumors were isolated using enzyme digestion and density gradient centrifugation. The resulting TIL fractions were then cloned in a limiting dilution assay (13) that allows virtualy all peripheral blood resting T cells to undergo clonal expansion. The frequency of proliferating T lymphocyte precursors (PTL-P) can then be calculated thus giving an indication of immunocompetent T cells present in the original infiltrate compared to patient and normal peripheral blood. In addition the clones obtained can be expanded to allow for functional assays in vitro. This report demonstrates that TIL cloned from human gliomas and grown in the presence of interleukin-2 do exhibit cytolytic activity in vitro in both allogeneic and autologous systems.
Key wordsTumor-infiltrating lymphocytes Limiting dilution analysis cytolytic activity
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