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Original Article

^a Ludwig Institute for Cancer Research, Brussels Branch, B-1200 Brussels, Belgium
^b Cellular Genetics Unit, Cliniques Universitaires St. Luc, Université Catholique de Louvain, B-1200 Brussels, Belgium
^c Medical Genetics Center, Cliniques Universitaires St. Luc, Université Catholique de Louvain, B-1200 Brussels, Belgium
^d Department of Nephrology, Cliniques Universitaires St. Luc, Université Catholique de Louvain, B-1200 Brussels, Belgium
^e Department of Urology, Cliniques Universitaires de Mont-Godinne, Université Catholique de Louvain, B-5530 Yvoir, Belgium
^f Department of Pneumology, Cliniques Universitaires de Mont-Godinne, Université Catholique de Louvain, B-5530 Yvoir, Belgium
Ludwig Institute for Cancer Research, Ave. Hippocrate 74, Université de Louvain 7459, B-1200 Brussels, Belgium.32-2-764-75-9032-2-764-75-72

vandeneynde{at}licr.ucl.ac.be

By stimulating blood lymphocytes from a renal cell carcinoma patient in vitro with the autologous tumor cells, we obtained cytolytic T lymphocyte (CTL) clones that killed several autologous and allogeneic histocompatibility leukocyte antigen (HLA)-B7 renal carcinoma cell lines. We identified the target antigen of these CTLs by screening COS cells transfected with the HLA-B7 cDNA and with a cDNA library prepared with RNA from the tumor cells. The antigenic peptide recognized by the CTLs has the sequence LPRWPPPQL and is encoded by a new gene, which we named RU2. This gene is transcribed in both directions. The antigenic peptide is not encoded by the sense transcript, RU2S, which is expressed ubiquitously. It is encoded by an antisense transcript, RU2AS, which starts from a cryptic promoter located on the reverse strand of the first intron and ends up on the reverse strand of the RU2S promoter, which contains a polyadenylation signal. This mechanism of antigen expression is unprecedented and further illustrates the notion that many peptides recognized by T cells cannot be predicted from the primary structure of the major product of the encoding gene. Antisense transcript RU2AS is expressed in a high proportion of tumors of various histological types. It is absent in most normal tissues, but is expressed in testis and kidney, and, at lower levels, in urinary bladder and liver. Short-term cultures of normal epithelial cells from the renal proximal tubule expressed significant levels of RU2AS message and were recognized by the CTLs. Therefore, this antigen is not tumor specific, but corresponds to a self-antigen with restricted tissue distribution.

Key Words: renal cell carcinoma • cytolytic T lymphocytes • antisense • peptides

Abbreviations used in this paper: RACE, rapid amplification of cDNA ends; RCC, renal cell carcinoma; RT, reverse transcription.

© 1999 The Rockefeller University Press

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