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In situ demonstration of renal-cell-carcinoma-specific T-cell clones

✍ Scribed by Anne Caignard; Maryvonne Guillard; Catherine Gaudin; Bernard Escudier; Frédéric Triebel; Pierre-Yves Dietrich

Publisher
John Wiley and Sons
Year
1996
Tongue
French
Weight
780 KB
Volume
66
Category
Article
ISSN
0020-7136

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✦ Synopsis

Using mixed lymphocyte tumor-cell culture (MLTC) in a selected renal-cell carcinoma, we derived a tumor-specific T-cell line in which Vf3 14+ and Vp 19+ T cells represented 70% of the whole T-cell population. Selected Vpl9+ T cells were CD8+ and exhibited a HLA-restricted specific cytotoxicity against tumor cells. Independently, 2 CTL clones were obtained by direct cloning of tumor-infiltrating lymphocytes, VlllC2 CTL expressing a Vpl9 and VllBlO CTL a Vp13 T-cell-receptor transcript. VllB I0 lysed autologous tumor cells, normal kidney cells and EBV-transformed B cells. In contrast, VlllC2 lysed tumor cells exclusively, demonstrating that the antigen structure recognized is tumor-specific. In addition, we used a PCR-based method to search for the presence of these CTL in situ. TCR f% chain of VlllC2 and VllB 10 CTL were sequenced and primers complementary to their N regions were synthesized. VlllC2 CTL constituted up to 60% of Val9 transcripts in MLTC T-cell lines derived from tumor-infiltrating lymphocytes, 23% in tumor and 26% in a tumor-draining lymph node, while VllBlO was not detected. Thus, VlllCZCTL was successfully derived from lymphocytes infiltrating a renal-cell carcinoma by direct cloning as well as by MLTC, probably because it was highly expanded in viva within the tumor composing almost 2% of the TIL.

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