HLA restriction and T‐cell‐receptor Vβ gene expression of cytotoxic T lymphocytes reactive with human squamous‐cell carcinoma of the head and neck

A human cytotoxic‐T‐lymphocyte (CTL) line capable of killing autologous tumor (AuTu) cell targets was established from peripheral‐blood lymphocytes of a patient with squamous‐cell carcinoma of the tongue. The cultured CTL were CD3+CD8+CDIIb‐HLA‐DR+T cell receptor (TCR) α/β+. When tested in 4‐hr 51Cr‐release assays against various lines of squamous‐cell carcinoma of the head and neck (SCCHN) and a variety of non‐squamous human tumor and normal cell targets, the CTL were found to lyse the autologous SCCHN cell line (PCI50) and 7 allogeneic SCCHN lines: PCI‐1, ‐2, ‐4A, ‐4B, ‐13, ‐30 and ‐38. Of these tumor cell lines, PCI‐13, ‐30 and ‐38 shared HLA‐A2 locus with the AuTu, PC1‐50, while PCI‐4A and ‐4B shared HLA‐B44 with AuTu. Lysis of AuTu (A2+B44+), PCI‐13 (A2+B44‐) and PCI‐4B (A2‐ B44+) by the CTL was efficiently inhibited by monoclonal antibodies (MAbs) to CD3, CD8, TCRα/β or the major‐histocompatibility‐complex(MHC)‐class‐l antigens. MAbs to HLA‐A2 antigens inhibited lysis of PCI‐50 or PCI‐13 targets by the CTL. In cold‐target inhibition assays, unlabeled PCI‐4B or PCI‐13 cells inhibited CTL lysis of AuTu targets. The CTL incubated in the presence of the HLA‐A2+ SCCHN PCI‐50 or ‐13, but not an HLA‐A2+ gastric carcinoma, produced TNF‐α, IFN‐γ and GM‐CSF. The CTL were tested for their TCR Vβ gene expression by polymerase chain reaction (PCR). At week 10 in culture, the time of the highest AuTu cytotoxicity mediated by the CTL line, Vβ was expressed by 26% of T cells. Three clones, obtained by limiting dilution from 10‐week CTL and selected for high cytotoxicity against AuTu, were found to be Vβ+. Further analysis of the specificity of these clones indicated lytic activity against PCI‐13 (A2+B44‐), but not PCI‐4B (A2‐B44+) targets. In 16‐week cultures, which retained AuTu cytotoxicity as well as Vβ expression, TCR Vβ2 was also expressed at high frequency (29%), and AuTu‐reactive clones were found to be Vβ2+. Our results indicate that at least 2 different CTL populations (Vβ2+ and β2+) are able to recognize SCCHN‐associated antigen(s) and that the Vβ6+ T cells are HLA‐A2 restricted, while Vβ6+ T cells are HLA‐A2 restricted, while Vβ2+ T cells may be HLA‐ B44 restricted. © 1994 Wiley‐Liss, Inc.