Published in Cancer Detection and Prevention 2000; 24(Supplement 1).

Clonal expansion of T cells in TIL and autologous PBL stimulated with ovarian cancer cell

K Hayashi MD, K Yonamine PhD, T Iida MD

Dept Obstetrics and Gynecology, Yokohama City Seibu Hospital, St Marianna University, Yokohama Kanagawa, Japan

AIM: Within human solid tumors, there are numbers of tumor infiltrating lymphocyte (TIL) such as T cells. However, it was not clear whether tumor-antigen-specific immune response was occurring within the solid tumor in vivo. Therefore, a system needed to be analyzed the TIL clonality of a tumor-antigen-specific response in vivo. METHODS: We tried to clonally expanded T cells directly in vivo, using the RT-PCR-SSCP method to analyze CDR3(V-D-J-C) regions of TCR (antigen recognition regions). In the tumor, the T cell clone accumulation was shown at almost all families regardless of VB. 1.We analyzed autologous TIL stimulated with autologous cancer cell to find out if the number of tumor-antigen-specific T cell clone expanded in this TIL. 2.Autologous peripheral blood lymphocytes (PBL) from ovarian tumor-bearing patient and autologous cancer cell line were co-cultured for 3 weeks. RESULTS: 1.After the culturing, we determined the increased tumor-antigen-specific immune response at the clonal level in TIL. By adding initially obtained tissue each week to further stimulate autologous TIL and extending culturing for 4weeks. A specific part of accumulated T cell clones was proliferated, while the remaining parts restricted, thus confirming the expansion of specific clones. 2.The induced T cell clones in PBL with the autologous tumor stimulation were identical in specific VB to an accumulated T clones in TIL, and the nucleotide sequencing study of tumor-specific clones in the stimulated PBL and in TIL also revealed the same result. CONCLUSION: The successful achievement was made in expanding T cell clonal accumulation in PBL with the autologous tumor stimulation that is identical with TIL.

KEY WORDS: tumor-specific T cell clone, ovarian cancer.

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Paper presented at the International Symposium on Impact of Biotechnology on Cancer Diagnostic & Prognostic Indicators; Geneva, Switzerland; October 28 - 31, 2000; in the section on immunotherapy.