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Abstract
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-gamma 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 language | English |
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Pages (from-to) | 2110-2121 |
Number of pages | 12 |
Journal | European Journal of Immunology |
Volume | 35 |
Issue number | 7 |
DOIs | |
Publication status | Published - 1 Jul 2005 |
Keywords
- adoptive immunotherapy
- T cells
- mini-LCL
- cytomegalovirus
- mini-EBV
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Dive into the research topics of 'Selection of CMV-specific CD8(+) and CD4(+) T cells by mini-EBV-transformed B cell lines'. Together they form a unique fingerprint.Projects
- 1 Finished
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The Development of Cellular Therapy for the Correction of CMV-Specific Immunodeficiency After Stem Cell Transplantation
Cobbold, M.
1/07/03 → 31/07/09
Project: Research Councils