Selection of CMV-specific CD8+ and CD4+ T cells by mini-EBV-transformed B cell lines

Martina Wiesner, Caroline Zentz, Markus H. Hammer, Mark Cobbold, Florian Kern, Hans Jochem Kolb, Wolfgang Hammerschmidt, Reinhard Zeidler, Andreas Moosmann

Research output: Contribution to journalArticlepeer-review

24 Scopus citations


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.

Original languageEnglish
Pages (from-to)2110-2121
Number of pages12
JournalEuropean Journal of Immunology
Issue number7
StatePublished - Jul 2005
Externally publishedYes


  • Adoptive immunotherapy
  • Cytomegalovirus
  • Mini-EBV
  • Mini-LCL
  • T cells


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