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In situ demonstration of renal‐cell‐carcinoma‐specific T‐cell clones
Author(s) -
Caignard Anne,
Guillard Maryvonne,
Gaudin Catherine,
Escudier Bernard,
Triebel Frédéric,
Dietrich PierreYves
Publication year - 1996
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/(sici)1097-0215(19960516)66:4<564::aid-ijc23>3.0.co;2-6
Subject(s) - ctl* , tumor infiltrating lymphocytes , cytotoxic t cell , biology , cd8 , t cell receptor , cell culture , t lymphocyte , microbiology and biotechnology , t cell , lymph node , antigen , cancer research , immunology , immune system , in vitro , biochemistry , genetics
Using mixed lymphocyte tumor‐cell culture (MLTC) in a selected renal‐cell carcinoma, we derived a tumor‐specific T‐cell line in which Vβ14 + and Vβ19 + T cells represented 70% of the whole T‐cell population. Selected Vβ19 + 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, VIIIC2 CTL expressing a Vβ19 and VIIB10 CTL a Vβ13 T‐cell‐receptor transcript. VIIB10 lysed autologous tumor cells, normal kidney cells and EBV‐transformed B cells. In contrast, VIIIC2 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 β chain of VIIIC2 and VIIB10 CTL were sequenced and primers complementary to their N regions were synthesized. VIIIC2 CTL constituted up to 60% of Vβ19 transcripts in MLTC T‐cell lines derived from tumor‐infiltrating lymphocytes, 23% in tumor and 26% in a tumor‐draining lymph node, while VIIB10 was not detected. Thus, VIIIC2CTL 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 vivo within the tumor composing almost 2% of the TIL. © 1996 Wiley‐Liss, Inc.