We have recently reported that tumor reactive T cell lines and clones can be developed from lymphocytes infiltrating ovarian malignant tumors and ascites (TIL/TAL). These cells are currently used for immunotherapy of ovarian cancer. In this study we investigated the presence of common non-germline rearrangements at TCR Cbeta1 among freshly isolated and cultured ovarian TIL/TAL. The cultured ovarian TIL/TAL show a characteristic T cell phenotype (>95% CD3+) and mediate tumor target lysis. Our results show that non-germline rearrangements can be detected in cultured ovarian TAL isolated from ascites but not in freshly isolated TAL. Certain of rearranged bands were common among cultured TAL. In contrast, in samples of freshly isolated TIL from solid ovarian tumors localized at different organ sites, common rearrangements were found among fresh and cultured TIL. The results suggest that the observed polyclonality of T cells isolated from freshly collected ascitic tumors may be due, at least in part, to either dilution by passenger lymphocytes, or the lack of an in vivo oligoclonal response to autologous tumor, and indicate the usefulness of molecular analysis of TCR V gene segments in T cells used for immunotherapy.

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