Ex vivo expansion of tumor-draining lymph node cells using compounds which activate intracellular signal transduction. II. Cytokine production and in vivo efficacy of glioma-sensitized lymphocytes

摘要: We have investigated the anti-tumor activity of ex vivo activated and expanded T cells which had been sensitized in to one two different syngeneic rat glioma cell lines; D74 or RT-2. Rats were by inoculation irradiated tumor into each hind foot pad. After 10 days, tumor-draining lymph node (DLN) from popliteal region was excised prepared as a single suspension. Tumor-DLN lymphocytes next overnight RPMI-1640 medium containing 10% fetal bovine serum (FBS), Bryostatin-1 (5 nM), ionomycin (1 μM), 20 U human recombinant interleukin-2 (IL-2) per ml. Culture for seven days supplemented with FBS IL-2 resultedin approximately 100-fold expansion lymphocyte population. Both D74- RT-2-sensitized constitutively secreted necrosis factor-α, both populations produced comparable amounts cytokine when co-cultured either gliomacell line. Neither effectors γ-interferon (γ-IFN), but γ-IFN exposed line vitro. D74-sensitized released significantly more than RT-2 DLN lymphocytes. In vitro 51Chromium-release assays indicated that cytotoxic targetsthan werealso targets. To assess therapeutic efficacy, rats who inoculated intradermally three earlier received an intravenous injection RT-2- (106Q cells/gram body weight) after activation equal number lymphokine-activated killer (LAK) cells. Tumordiameters measured daily revealed glioma- led elimination while treatmentwith LAK no benefit. These results indicate, at least these lines, release, rather cytotoxicity, better predictor immunotherapeutic efficacy glioma-sensitized,