HLA-DR-antigen-associated restriction of EBV-specific cytotoxic T-cell colonies

Abstract

EBV-specific cytotoxic T cells can be generated in vitro in a secondary response. Several previous studies with bulk cultures provided evidence that cytotoxicity was restricted by HLA-A,B-related antigens. In the present family study, the EBV-specific cytotoxic T-cell response of a normal EBV-seropositive donor was analysed in detail by T-cell colony formation. Peripheral blood mononuclear cells were stimulated by the gamma-irradiated autologous lymphoblastoid cell line (LCL) and 3 days later seeded into agarose. Colonies were harvested, amplified by addition of interleukin-2 (IL-2), and analysed for T-cell markers and specificity in 51Cr-release assays. Twenty-two colonies were studied: all colonies were OKT3+, five were predominantly OKT4+, 9 were OKT8+ and 8 were mixtures. As expected from previous work, the OKT8+ colonies were cytotoxic for the autologous LCL target and cytotoxicity was blocked by monoclonal antibody (W6/32) to the nonpolymorphic determinants of HLA-A,B,C antigens. Significantly, the OKT4+ colonies tested also showed specific cytotoxicity, but lysis of the autologous LCL was blocked by the monoclonal antibody (OKlal) to the non-polymorphic determinants of HLA-DR antigens. Two interesting patterns of specificity were seen in cytotoxicity tests on sibling LCL targets. In one pattern, targets bearing the A11, B5, DR7 haplotype were lysed, while those bearing the A1, B8, DR3 were not, indicating haplotype preference. In the other pattern, there was lysis of the autologous cell line but not of the sibling targets. These results including HLA-DR-associated restriction, haplotype preference and strict self-preference, further illustrate the complexity of the EBV-cytotoxic T-cell response.

Failure of Epstein-Barr virus-specific cytotoxic T lymphocytes to lyse B cells transformed with the B95-8 strain is mapped to an epitope that associates with the HLA-B8 antigen

“…Both the FCS-and HS-grown autologous LCL were lysed equally well by the EBV-specilk CTL in keeping wiih previous findings thai virus-specific lysis is independent of scrum effects [19]. Second, the haploidentieal LCL target cells (donors 4,5,8) bearing the HLA-AI, -B8, -DR3 paternal haplotype were not lysed by the specific CTL (Fig. !b. c) even though this haplolype was expressed by the autologous LCL used in the initial stimulation.…”

CD4+ T-cell clones recognizing human lymphoma-associated antigens: generation by in vitro stimulation with autologous Epstein-Barr virus–transformed B cells

“…A review of earlier literature shows that CD4 ϩ T cells capable of recognizing autologous LCL cells have been detected in various in vitro settings, usually after LCL stimulation, where the effector cells were derived from either fetal cord blood, 36,37 EBVseronegative donors, 35,38 EBV-seropositive donors, [39][40][41][42][43] or, most recently, patients with infectious mononucleosis undergoing primary EBV infection. 21 In some cases, further analysis at the clonal level showed these cells to be HLA II restricted or to recognize LCL cells but not mitogen-activated lymphoblasts.…”

“…21 In some cases, further analysis at the clonal level showed these cells to be HLA II restricted or to recognize LCL cells but not mitogen-activated lymphoblasts. 21,38,[40][41][42] However, most of these studies did not look for reactivity against individual EBV antigens and so never showed EBV specificity. We suggest that many of these responses were, as described here, LCL specific.…”

Cytotoxic T lymphocyte discrimination between type A Epstein--Barr virus transformants is mapped to an immunodominant epitope in EBNA 3

“…Rosetted T cells were also maintained as polyclonal CTL lines by restimulation with recombinant interleukin-2 (rlL-2) and specific irradiated transformants. T cell clones and PHA blasts were generated as previously described (Misko et al, 1984;Burrows et al, 1990a).…”

“…Polyclonal CTLs from donor IM and T cell clones from donor LC were tested for cytotoxicity in standard 5 h SlCr-release assays (Misko et al, 1984). The target panel included the type A and type B autologous LCLs and autologous PHA blasts.…”