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Abstract: B cells immortalized by a mini-Epstein-Barr virus encoding a foreign antigen efficiently reactivate specific cytotoxic T cells

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www.ProHealth.com • August 19, 2002


Moosmann A, Khan N, Cobbold M, Zentz C, Delecluse HJ, Hollweck G, Hislop AD, Blake NW, Croom-Carter D, Wollenberg B, Moss PA, Zeidler R, Rickinson AB, Hammerschmidt W.

Department of Otorhinolaryngology, Ludwig-Maximilians-Universitat, Munich, Germany; GSF, Institute for Clinical Molecular Biology and Tumor Genetics, Department of Gene Vectors, and Clinical Cooperation Group Molecular Oncology, Munich, Germany; Vaecgene Biotech, Munich, Germany; and the Cancer Research UK Institute for Cancer Studies, University of Birmingham, Birmingham, United Kingdom.

Lymphoblastoid cell lines (LCLs) are human B cells latently infected and immortalized by Epstein-Barr virus (EBV). Presenting viral antigens, they efficiently induce EBV-specific T-cell responses in vitro. Analogous ways to generate T-cell cultures specific for other antigens of interest are highly desirable. Previously, we constructed a mini-EBV plasmid that consists of less than half the EBV genome, is unable to cause virus production, but still immortalizes B cells in vitro. Mini-EBV-immortalized B-cell lines (mini-LCLs) are efficiently produced by infection of B cells with viruslike particles carrying only mini-EBV DNA. Mini-EBV plasmids can be engineered to express an additional gene in immortalized B cells.

Here we present a mini-EBV coding for a potent CD8(+) T-cell antigen, the matrix phosphoprotein pp65 of human cytomegalovirus (CMV). By means of this pp65 mini-EBV, pp65-expressing mini-LCLs could be readily established from healthy donors in a one-step procedure. We used these pp65 mini-LCLs to reactivate and expand effector T cells from autologous peripheral blood cells in vitro. When generated from cytomegalovirus (CMV)-seropositive donors, these effector T-cell cultures displayed strong pp65-specific HLA-restricted cytotoxicity.

A large fraction of CD8(+) T cells with pp65 epitope specificity was present in such cultures, as demonstrated by direct staining with HLA/peptide tetramers. We conclude that the pp65 mini-EBV is an attractive tool for CMV-specific adoptive immunotherapy. Mini-EBVs could also facilitate the generation of T cells specific for various other antigens of interest. (Blood. 2002;100:1755-1764)

PMID: 12176897 [PubMed - as supplied by publisher]




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