Cytotoxic T cell recognition of Epstein‐Barr virus‐infected B cells. III. Establishment of HLA‐restricted cytotoxic T cell lines using interleukin 2

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Abstract
Epstein‐Barr virus (EBV)‐specific cytotoxic T cell precursors, present in the circulation of previously infected (seropositive) individuals, have been reactivated in vitro by challenging with autologous EBV‐transformed cells, and the reactivated populations subsequently expanded as interleukin 2 (IL2)‐dependent cell lines. These lines were dominated by T cells possessing the cytotoxic/suppressor cell surface phenotype and, when tested for effector function in chromium‐release assays, demonstrated potent EBV‐specific, HLA‐A and ‐B antigen‐restricted cytotoxicity even when derived from seropositive donors whose initial cytotoxic response to in vitro reactivation was relatively weak. With all the lines tested from 10 seropositive donors, strong killing of autologous EBV‐transformed cells was observed in the absence of any significant lysis of autologous mitogen‐stimulated lymphoblasts or of a panel of EBV genome‐negative cell lines sensitive to natural killing. Furthermore, the availability of IL2‐expanded effectors cell populations allowed their being tested upon a wide panel of allogeneic EBV‐transformed targets such that the dominant HLA‐restricted reactivities within these populations could be identified. Monoclonal antibody blocking experiments confirmed that lysis of the autologous EBV‐transformed cell line by IL2‐expanded effectors could be specifically inhibited (a) by pretreatment of the target cells with antibodies binding to the HLA/β2‐microglobulin complex, and (b) by pretreatment of the effector cells with the cytotoxic/suppressor T cell‐specific antibody Leu 2a.

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