Analysis of Cytokine Secretion by Melanoma-Specific CD4+T Lymphocytes

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Abstract
Although specific antitumor immune reactivity has been documented extensively in CD8+ T cells derived from melanoma patients, relatively little is known about CD4+ T cell responses against melanoma. Tumor-infiltrating lymphocytes (TIL) cultured from metastatic lesions in five patients yielded cytolytic CD8+ T cells with specific activity against autologous and MHC class I-compatible allogeneic melanoma targets. In four of the five cases studied, CD4+ T cells purified from bulk TIL cultures also reacted specifically with autologous melanoma cells, as manifested by the secretion of various cytokines (GM-CSF, TNF-α, and IFN-γ) after a 24 h cocultivation. Cytokine secretion by CD4+ T cells was MHC class II restricted, and proved to be a more reliable indicator of the immunologic reactivity of CD4+ T cells than cytolysis. Three of the four reactive CD4+ TIL failed to recognize allogeneic melanomas, suggesting recognition of Ag with limited expression in the patient population. Cloning such Ags may provide clues to optimizing current antitumor immunization strategies.

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