Oelke, M. and Moehrle, U. and Chen, J. L. and Behringer, D. and Cerundolo, V. and Lindemann, A. and Mackensen, Andreas (2000) Generation and purification of CD8(+) Melan-A-specific cytotoxic T lymphocytes for adoptive transfer in tumor immunotherapy. CLINICAL CANCER RESEARCH, 6 (5). pp. 1997-2005. ISSN 1078-0432
Full text not available from this repository. (Request a copy)Abstract
Tumor antigens that might serve as potential targets for adoptive T-cell therapy have been defined in different tumor entities, especially in malignant melanoma, To generate conditions to induce primary T-cell responses against different HLA-A*0201-restricted melanoma peptides and to allow further expansion of peptide-specific T cells for adoptive transfer, CD8(+)-purified T cells from healthy donors mere stimulated with Melan-A-pulsed autologous dendritic cells. Dendritic cells were generated irt vitro from monocytes with granulocyte macrophage colony-stimulating factor, interleukin-l, and transforming growth factor-beta. After 3-4 weekly stimulation cycles with Melan-A-pulsed DCs, we were able to induce a strong peptide-specific CTL response in vitro. MHC-peptide tetramer staining revealed a frequency of up to 3.5% CD8+/Melan-A(+) T cells. Additional antigen-independent expansion with anti-CD3/anti-CD28 monoclonal antibodies together with interleukin-2 gave rise to 600-fold expansion of CD8(+) CTLs that maintained Melan-A specificity and were able to efficiently lyse Melan-A-expressing melanoma cells. To enrich antigen-specific T cells in vitro, me used a recently established technology for analysis and sorting of live cells according to secreted cytokines, In the present study, we demonstrated that Melan-A-specific T cells can be purified by magnetic separation according to secreted IFN-gamma, These cells revealed a very potent monospecific CTL response, even at low E:T ratios, against Melan-A-pulsed and Melan-A-expressing target cells. Altogether, our study demonstrated that me have developed an efficient method for generating large numbers of peptide-specific T cells iii vitro that mag be used for adoptive T-cell transfer in tumor immunotherapy.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | CELLS IN-VITRO; INFILTRATING LYMPHOCYTES; METASTATIC MELANOMA; PERIPHERAL-BLOOD; HLA-A2 MELANOMAS; PROGENITOR CELLS; DENDRITIC CELLS; BIRBECK GRANULE; REACTIVE CTL; BETA-CHAIN; |
| 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: | 03 May 2022 14:18 |
| Last Modified: | 03 May 2022 14:18 |
| URI: | https://pred.uni-regensburg.de/id/eprint/42502 |
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