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The T-Cell Receptor Vß Gene Usage in Tumor-infiltrating Lymphocytes and Blood of Patients with Hepatocellular Carcinoma

Departments of Pathology [E. W., T. L. W., R. B. H., M. T.], Medicine [R. B. H.], and Pediatrics [R. G., M. T.], University of Pittsburgh School of Medicine and Pittsburgh Cancer Institute [E. W., T. L. W., R. G., R. B. H.], Pittsburgh, Pennsylvania 15213

To determine a possibly restricted T-cell receptor (TCR) repertoire in tumor-infiltrating lymphocytes (TIL) in response to tumor-associated antigens in patients with hepatocellular carcinoma (HCC), freshly isolated TIL (n = 5) and peripheral blood lymphocytes (PBL; n = 6; 3 paired with TIL) were studied for expression of TCR variable (V) ß regions. RNA purified from TIL or PBL was reverse-transcribed into complementary DNA. This complementary DNA was amplified by quantitative polymerase chain reaction with 22 primers specific for 20 TCR Vß gene families and a 3' constant (C) ß primer. As a reference for later quantitation, a fragment of TCR C was coamplified with each Vß region. Using ³²P-labeled 3' primers, the percentage of total Vß expression was calculated by measuring the cpm of each of the amplified products. In contrast to PBL of 6 control, healthy individuals, whose range of expression of each TCR Vß gene varied from 0 to 13%, the expression of some Vß genes in HCC TIL was as high as 33%, indicating a restricted TCR Vß usage in HCC TIL. When polymerase chain reaction-amplified complementary DNAs of the Vß1 or Vß3 genes obtained from two TIL preparations were cloned and sequenced, the same rearrangements were found in the majority of DNA clones. The particular Vß genes that were over- or underrepresented in TIL varied among the patients. In 3 of 6 PBL and 3 of 5 TIL, the Vß3 gene was expressed with a relatively high frequency. The Vß4 gene expression was consistently low in patients' TIL or PBL. In 3 paired PBL and TIL, Vß expression was similar. In 5 of 6 cases, HCC PBL had different TCR Vß frequencies from those seen in normal PBL. This analysis of TCR Vß usage in freshly isolated TIL and in PBL indicated that T-lymphocytes in patients with HCC might have restricted immunological reactivity and that Vß3-restricted TIL might represent antitumor effector cells.

¹ Supported by a grant from Deutsche Forschungsgemeinschaft (We1335/1-1) and Grant ACS-IRG IN-5830. Supported in part by NIH Project PO1-CA47445.

² To whom requests for reprints should be addressed, at Pittsburgh Cancer Institute, Room W1041, Biomedical Science Tower, DeSoto at O'Hara Street, Pittsburgh, PA 15213.

Received 2/28/92. Accepted 8/24/92.

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