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- Title
Functional Characterization of CD8<sup>+</sup> Antigen-Specific Cytotoxic T Lymphocytes after Enrichment Based on Cytokine Secretion: Comparison with the MHC-Tetramer Technology.
- Authors
Oelke, M.; Kurokawa, T.; Hentrich, I.; Behringer, D.; Cerundolo, V.; Lindemann, A.; Mackensen, A.
- Abstract
Cell therapy with antigen-specific T cells holds promise for various diseases including cancer and viral infections. The powerful enrichment procedure based on major histocompatibility complex (MHC)-tetramers, however, is of limited applicability so far. Therefore, the recently developed cell surface affinity matrix technology that allows direct identification and enrichment of life antigen-specific T cells based on cytokine secretion was evaluated in this respect. To this end, CD8+ T cells directed against the HLA-A*0201-restricted melanoma-associated peptide Melan-A (aa26–35) were generated by combining stimulation of peptide-pulsed autologous dendritic cells (DC) with antigen-independent expansion with anti-CD3/anti-CD28 monoclonal antibodies (MoAb). Antigen-specific cytotoxic T lympocyte (CTL) were detected based on stimulation-induced interferon (IFN)-γ and interleukin (IL)-4 secretion and enriched > 100-fold using the cell surface affinity matrix technology. The resulting IFN-γ- and IL-4-secreting CTL lines contained > 80% and > 70% cytokine positive T cells, respectively. They exhibited a cytotoxic activity against Melan-A expressing target cells that was significantly higher as compared to nonpurified CTL. Direct staining of enriched CTL with HLA-A2-Melan-A-tetramers revealed a high correlation between the results obtained from the cell surface affinity matrix technology and those obtained from tetrameric complexes. Altogether, our study demonstrates that cytokine-driven enrichment based on the cell surface affinity matrix technology enables selective isolation of functionally active antigen-specific CTL that may be used for an adoptive T cell transfer in immunotherapy.
- Publication
Scandinavian Journal of Immunology, 2000, Vol 52, Issue 6, p544
- ISSN
0300-9475
- Publication type
Academic Journal
- DOI
10.1046/j.1365-3083.2000.00810.x