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 Blood, 23 July 2009, Vol. 114, No. 4, pp. 807-815.
Prepublished online as a Blood First Edition Paper on May 14, 2009; DOI 10.1182/blood-2008-12-194043.

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IMMUNOBIOLOGY

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

Heather M. Long1, Jianmin Zuo1, Alison M. Leese1, Nancy H. Gudgeon1, Hui Jia1, Graham S. Taylor1, and Alan B. Rickinson1

1 Cancer Research UK (CRUK) Institute for Cancer Studies, School of Cancer Sciences, University of Birmingham, Birmingham, United Kingdom

Epstein-Barr virus (EBV)–specific T-cell preparations, generated by stimulating immune donor lymphocytes with the autologous virus-transformed B-lymphoblastoid cell line (LCL) in vitro, can be used to target EBV-positive malignancies. Although these preparations are enriched for EBV antigen–specific CD8+ T cells, most also contain a CD4+ T-cell population whose specificity is unknown. Here, we show that, although CD4+ T-cell clones derived from such cultures recognize HLA class II–matched LCLs but not mitogen-activated B lymphoblasts, many (1) do not map to any known EBV antigen, (2) can be raised from EBV-naive as well as EBV-immune persons, and (3) can recognize a broad range of human B lymphoma–derived cell lines irrespective of EBV genome status, providing those lines to express the relevant HLA class II–restricting allele. Importantly, such CD4+ clones not only produce IFN{gamma} but are also cytotoxic and can control the outgrowth of HLA-matched lymphoma cells in cocultivation assays. We infer that such CD4+ T cells recognize cellular antigens that are preferentially up-regulated in EBV-transformed but not mitogen-activated B lymphoblasts and that are also expressed in a range of B-cell malignancies. Such antigens are therefore of potential value as targets for CD4+ T cell–based immunotherapy.


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