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Functional Characterization of CD8 + Antigen‐Specific Cytotoxic T Lymphocytes after Enrichment Based on Cytokine Secretion: Comparison with the MHC‐Tetramer Technology
Author(s) -
Oelke M.,
Kurokawa T.,
Hentrich I.,
Behringer D.,
Cerundolo V.,
Lindemann A.,
Mackensen A.
Publication year - 2000
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.2000.00810.x
Subject(s) - cytotoxic t cell , ctl* , antigen , biology , major histocompatibility complex , cd8 , t cell , microbiology and biotechnology , cd28 , antigen presenting cell , immunotherapy , immunology , immune system , biochemistry , in vitro
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.