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Melanoma-reactive human cytotoxic T lymphocytes derived from skin biopsies of delayed-type hypersensitivity reactions induced by injection of an autologous melanoma cell line.

The expression by melanomas of multiple antigens that are recognized by specific MHC class I-restricted CTLs has been clearly demonstrated. The goal of many immunotherapy protocols being developed is, therefore, the induction and/or augmentation of CTLs specific for such antigens. One approach has been to immunize using irradiated autologous melanoma cells. Responses to this type of immunization and others are often subsequently measured by delayed-type hypersensitivity (DTH) reactions. The aim of this work was to characterize whether specific CTL responses occur at such DTH sites. Cutaneous DTH reactions were observed following injection of irradiated autologous melanoma cells expressing known tumor antigens. We isolated lymphocytes from biopsies of DTH reaction sites and could measure melanoma-specific CTL activity after 2-3 weeks of culture. The T-cell receptor-Vbeta repertoire of the cultured lymphocytes, assessed by flow cytometry, was highly skewed in both the CD4(+) and CD8(+) T-cell subsets. The repertoires were different among cultures derived from independent biopsies of simultaneous or subsequent DTH reaction sites and very different to that of fresh peripheral blood lymphocytes (PBLs) or PBLs cultured under the same conditions. No particular T-cell expansions dominated several DTH reaction sites, nor could they be detected in PBLs. It appears that T-cell responses to this type of immunization may be limited to the local microenvironment. Establishing the value of DTH reactions in determining levels of systemic antitumor immunity requires further investigation; however, such reactions may indicate a patient's competence to mount an antitumor immune response and enable the isolation of tumor-specific CTLs for use in tumor antigen identification.