title: Lymphocytes infiltrating human ovarian tumors: synergy between tumor necrosis factor alpha and interleukin 2 in the generation of CD8+ effectors from tumor-infiltrating lymphocytes.
abstract: Tumor-infiltrating lymphocytes (TIL) were isolated by enzymatic digestion and gradient centrifugation from 18 human ovarian carcinomas. These cells were cultured in a complete medium supplemented with recombinant interleukin 2 (IL2) alone or recombinant IL2 plus recombinant tumor necrosis factor alpha (TNF-alpha), and their growth and antitumor cytotoxicity were determined. TIL cultured in the presence of IL2 plus TNF-alpha (1000 units/ml each) for 6 days showed significantly higher cytotoxicity against fresh autologous tumor targets than did TIL cultured with IL2 alone (e.g., mean lytic units/10(7) cells for 8 TIL preparations were 290 versus 74; P less than 0.05). No differences in [3H]thymidine uptake or natural killer cell activity were observed among these TIL cultures. In titration experiments, optimal synergistic concentrations of IL2 and TNF-alpha were determined as 10(2) and 10(3) units/ml, respectively. Using these concentrations for culturing the TIL, effector cells developed which preferentially lysed autologous tumor and displayed a CD8+ phenotype (up to 75% positive). However, the autologous tumor cytotoxicity mediated by these cultured TIL on day 6 was short lived. By day 12, it was replaced by non-major histocompatibility complex-restricted, lymphokine-activated killer cell-like activity mediated by CD3-CD56+ effector cells. Simultaneously, the production of gamma-interferon and interleukin 1 decreased in these cultures. In contrast to TNF-alpha, anti-CD3 antibody synergized with IL2 to increase 2-3-fold TIL proliferation but not their cytotoxic activity against autologous tumor cell targets. These data suggest that TNF-alpha and IL2 synergize early in culture to induce tumor-reactive CD8+ effectors, some of which may be specific for autologous ovarian tumor cells. However, the conditions needed to sustain the specific autologous tumor responses in long-term cultures of human TIL remain to be determined.
authors.affiliation: Department of Pathology, University of Pittsburgh School of Medicine 15261.
authors.forename: Y L
authors.initials: YL
authors.lastname: Wang
authors.forename: L S
authors.initials: LS
authors.lastname: Si
authors.forename: A
authors.initials: A
authors.lastname: Kanbour
authors.forename: R B
authors.initials: RB
authors.lastname: Herberman
authors.forename: T L
authors.initials: TL
authors.lastname: Whiteside
mesh_terms: D000945:Antigens, Differentiation, T-Lymphocyte; D016827:CD8 Antigens; D002469:Cell Separation; D003602:Cytotoxicity, Immunologic; D005260:Female; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology*; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000187:drug effects / Q000276:immunology* / Q000473:pathology; D010051:Ovarian Neoplasms / Q000276:immunology* / Q000473:pathology; D011994:Recombinant Proteins / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
chemical_list: D000945:Antigens, Differentiation, T-Lymphocyte; D016827:CD8 Antigens; D007376:Interleukin-2; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha