Characterization of OKT3-initiated lymphokine-activated effectors expanded with interleukin 2 and tumor necrosis factor alpha.
Synergistic and cooperative effects in vitro of OKT3, interleukin 2 (IL-2), and tumor necrosis factor alpha (TNF) as stimuli in generating effectors with lymphokine-activated killer activity were studied. Activation of human peripheral blood mononuclear cells with OKT3 (10 ng/ml) for 48 h, followed by culture in low concentrations of IL-2 (10 units/ml) and TNF (250 units/ml) resulted in higher cell recovery (50- to 3300-fold) compared to the number of cells in the initial culture and enhanced lytic activity against both Raji and fresh lung tumor targets (mean 100-fold) by day 30 compared to those expanded with higher concentrations of IL-2 (100 units/ml) alone. Immunofluorescence analysis of peripheral blood mononuclear cells initiated with OKT3 and expanded with IL-2 plus TNF revealed a selective increase in CD8+ cells and a decrease in CD4+ by day 28; the opposite effect was observed when cells were incubated with 100 units/ml of IL-2 alone, resulting in a greater proportion of CD4+ cells. Almost all cells were CD3+. Studies of cytokine receptor expression indicated that OKT3 plus IL-2 plus TNF caused an earlier up-regulation of the IL-2 receptor beta chain (Tac) and higher TNF receptor expression by day 6 compared to 100 units/ml IL-2 alone. Significant TNF levels (greater than 17 units/ml) were measured in culture supernatants from peripheral blood mononuclear cells initiated with OKT3 alone. Collectively, our data demonstrate that induction of lymphokine-activated killer activity with OKT3, followed by culture in low concentrations of IL-2 plus TNF is an alternative to the use of high-dose IL-2 alone and suggest that this combination may provide potential advantages in long-term generation of cytolytic cells.
Yang
S C
SC
Department of Tumor Biology, University of Texas M. D. Anderson Cancer Center, Houston 77030.
Owen-Schaub
L B
LB
Roth
J A
JA
Grimm
E A
EA
D000945:Antigens, Differentiation, T-Lymphocyte / Q000494:pharmacology*; D017252:CD3 Complex; D002470:Cell Survival / Q000187:drug effects; D004357:Drug Synergism; D006801:Humans; D007167:Immunotherapy; D007376:Interleukin-2 / Q000494:pharmacology*; D015979:Killer Cells, Lymphokine-Activated / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D007958:Leukocyte Count; D008175:Lung Neoplasms / Q000628:therapy; D008213:Lymphocyte Activation / Q000187:drug effects*; D010641:Phenotype; D011948:Receptors, Antigen, T-Cell / Q000494:pharmacology*; D011956:Receptors, Cell Surface / Q000096:biosynthesis; D015375:Receptors, Interleukin-2 / Q000096:biosynthesis; D018124:Receptors, Tumor Necrosis Factor; D013997:Time Factors; D014407:Tumor Cells, Cultured / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000494:pharmacology*
D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D007376:Interleukin-2; D011948:Receptors, Antigen, T-Cell; D011956:Receptors, Cell Surface; D015375:Receptors, Interleukin-2; D018124:Receptors, Tumor Necrosis Factor; D014409:Tumor Necrosis Factor-alpha