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Open Access Green as soon as Postprint is submitted to ZB.
Selection of CMC-specific CD8⁺ and CD4⁺ T cells by mini-EBV-transformed B cell lines.
Eur. J. Immunol. 35, 2110-2121 (2005)
Efficient protocols to generate cytomegalovirus (CMV)-specific T cells are required for adoptive immunotherapy. Recombinant Epstein-Barr virus (EBV) vectors called mini-EBV can be used to establish permanent B cell lines in a single step, which present the CMV antigen pp65 in a constitutive manner. These B cell lines, coined pp65 mini-LCL, were successfully used to reactivate and expand CMV-specific cytotoxic T cells. Here we evaluate this pp65 mini-EBV system in closer detail, focusing on (1) the quantification of T cells with specific effector function and (2) the identification of CMV-specific CD4+ helper T cells. The co-expansion of various functional CMV epitope specificities was demonstrated by IFN-γ enzyme-linked immunospot assay (ELISPOT) assays and HLA-peptide tetramer staining. Single-cell cloning resulted in both CD4+ and CD8+ T cell clones, the majority of which was CMV specific. Thus, mini-LCL present the pp65 antigen on HLA class I and II, mobilizing both arms of the T cell response. Using a peptide library covering the pp65 sequence for further analysis of T cell clones, we identified new pp65 CD8+ and CD4+ T cell epitopes.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
cytomegalovirus; T cells; mini-EBV; mini-LCL; adoptive immunotherapy
ISSN (print) / ISBN
0014-2980
e-ISSN
1521-4141
Journal
European Journal of Immunology
Quellenangaben
Volume: 35,
Issue: 7,
Pages: 2110-2121
Publisher
Wiley
Publishing Place
Hoboken
Reviewing status
Peer reviewed
Institute(s)
Research Unit Gene Vector (AGV)
CCG Hematopoetic Cell Transplants (IMI-KHZ)
CCG Hematopoetic Cell Transplants (IMI-KHZ)