Cytokine combinations for induction of antigen-specific cytolytic T lymphocytes from peripheral blood lymphocytes.
We studied effects of varied cytokine combinations on allogeneic cytotoxic T lymphocyte (CTL) generation in a mixed lymphocyte-tumor cell culture. Sensitized with allogeneic melanoma cell line (MM-8.1 or MM-28), peripheral blood mononuclear cells (PBMC) were cultured in medium containing various combinations of cytokines that included human recombinant interleukin-2 (rIL-2) alone (MB-2), rIL-2 and rIL-4 (MB-2,4) and rIL-1, rIL-2, rIL-4 and rIL-6 (MB-1,2,4,6). Lymphocytes proliferated for more than 50 days and manifested stimulating cell-specific cytotoxicities in 1 of 5 cultures with MB-2, in 4 of 5 with MB-2,4 and in 5 of 5 with MB-1,2,4,6. Lymphocytes grew up to 10(6) fold in culture with MB-2,4 or MB-1,2,4,6 at day 90-100. Stimulating cell MM-8.1 (HLAABC+,DR-) induced CD3+, CD8+, CD56- CTLs and MM-28 (HLA-ABC+, DR+) mainly generated CD3+, CD4+, CD56- CTLs. Monoclonal antibodies against HLA-ABC and HLA-DR molecules suppressed the stimulating cell-specific cytolytic activity of CD8+ and CD4+ CTLs, respectively. These data indicate that combination of rIL-1, rIL-2, rIL-4 and rIL-6 offer an efficient system to generate allogeneic, tumor-specific CTLs from PBMCs and that HLA molecules expressed on stimulating cells play an important role in CTL generation.
Tsujitani
S
S
Wistar Institute, Philadelphia, Pennsylvania 19104-4268, USA.
Nakashima
M
M
Watanabe
T
T
Kaibara
N
N
Koprowski
H
H
Steplewski
Z
Z
D015703:Antigens, CD / Q000096:biosynthesis; D002455:Cell Division; D002460:Cell Line; D002478:Cells, Cultured; D016207:Cytokines / Q000494:pharmacology*; D003602:Cytotoxicity, Immunologic; D004347:Drug Interactions; D015234:HLA-A Antigens / Q000096:biosynthesis; D006684:HLA-DR Antigens / Q000096:biosynthesis; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology; D007376:Interleukin-2 / Q000494:pharmacology; D015847:Interleukin-4 / Q000494:pharmacology; D015850:Interleukin-6 / Q000494:pharmacology; D007700:Kinetics; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000187:drug effects / Q000276:immunology*; D008545:Melanoma; D011994:Recombinant Proteins / Q000494:pharmacology; D013602:T-Lymphocytes, Cytotoxic / Q000187:drug effects / Q000276:immunology*; D014407:Tumor Cells, Cultured
D015703:Antigens, CD; D016207:Cytokines; D015234:HLA-A Antigens; D006684:HLA-DR Antigens; D007375:Interleukin-1; D007376:Interleukin-2; D015850:Interleukin-6; D011994:Recombinant Proteins; D015847:Interleukin-4