Oelke, M. and Kurokawa, T. and Hentrich, I. and Behringer, D. and Cerundolo, V. and Lindemann, A. and Mackensen, Andreas (2000) Functional characterization of CD8(+) antigen-specific cytotoxic T lymphocytes after enrichment based on cytokine secretion: Comparison with the MHC-tetramer technology. SCANDINAVIAN JOURNAL OF IMMUNOLOGY, 52 (6). pp. 544-549. ISSN 0300-9475
Full text not available from this repository. (Request a copy)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)-gamma and interleukin (IL)-4 secretion and enriched > 100-fold using the cell surface affinity matrix technology. The resulting IFN-gamma- 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.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | IN-VITRO; PERIPHERAL-BLOOD; REACTIVE CTL; CELLS; MELANOMA; GENERATION; DIFFERENTIATION; CD4(+); GENE; |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Innere Medizin III (Hämatologie und Internistische Onkologie) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 15 Mar 2022 07:58 |
| Last Modified: | 15 Mar 2022 07:58 |
| URI: | https://pred.uni-regensburg.de/id/eprint/42036 |
Actions (login required)
 |
View Item |
