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.
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.
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使用激活细胞内信号转导的化合物体外扩增肿瘤引流淋巴结细胞。
DOI：
10.1023/a:1005771717409
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发表时间：
1997
影响因子：
3.9
通讯作者：
Merchant,RE
中科院分区：
文献类型：
--
作者：
Rice,CD;Baldwin,NG;Biron,RT;Bear,HD;Merchant,RE
我们已经研究了体外活化和扩增的T细胞的抗肿瘤活性，所述T细胞已经在体内对两种不同的同基因大鼠胶质瘤细胞系之一（D 74或RT-2）致敏。通过将辐照的肿瘤细胞接种到每个后足垫中来致敏大鼠。10天后，切除每个腘区的肿瘤引流淋巴结（DLN），并制备成单细胞悬液。接着，在每ml含有10%胎牛血清（FBS）、苔藓抑素-1（5 nM）、离子霉素（1 μM）和20 U人重组白细胞介素-2（IL-2）的RPMI-1640培养基中活化肿瘤-DLN淋巴细胞过夜。在添加FBS和IL-2的RPMI-1640中培养7天，淋巴细胞群扩增约100倍。D 74和RT-2致敏的T细胞组成性分泌肿瘤坏死因子-α，当与任一胶质细胞系共培养时，两种淋巴细胞群产生相当数量的细胞因子。D 74和RT-2致敏的效应子均不组成性分泌γ-干扰素（γ-IFN），但在体外暴露于任一胶质瘤细胞系时，两个群体均产生γ-IFN。D 74致敏的T细胞比RT-2 DLN淋巴细胞释放更多的γ-IFN。体外铬释放试验表明，RT-2致敏的T细胞对RT-2靶细胞的细胞毒性大于对D 74细胞系的细胞毒性，D 74致敏的效应细胞对RT-2靶细胞的细胞毒性也更大。为了评估体内治疗效果，三天前皮内接种RT-2细胞的大鼠接受静脉内注射RT-2或D 74致敏的DLN细胞（106 Q细胞/克体重），所述DLN细胞在用苔藓抑素-1和离子霉素活化后扩增，或等量的淋巴因子激活的杀伤（LAK）细胞。每天测量肿瘤直径，结果显示注射胶质瘤致敏淋巴细胞可消除肿瘤，而LAK细胞治疗无疗效。这些结果表明，至少对于这两个胶质瘤系，γ-IFN释放，而不是体外细胞毒性，是胶质瘤致敏的扩增的T细胞的体内免疫效力的更好的预测因子。
We have investigated the anti-tumor activity of ex vivo activated and expanded T cells which had been sensitized in vivo to one of two different syngeneic rat glioma cell lines; D74 or RT-2. Rats were sensitized by inoculation of irradiated tumor cells into each hind foot pad. After 10 days, the tumor-draining lymph node (DLN) from each popliteal region was excised and prepared as a single cell suspension. Tumor-DLN lymphocytes were next activated overnight in RPMI-1640 medium containing 10% fetal bovine serum (FBS), Bryostatin-1 (5 nM), ionomycin (1 μM), and 20 U human recombinant interleukin-2 (IL-2) per ml. Culture for seven days in RPMI-1640 supplemented with FBS and IL-2 resultedin approximately 100-fold expansion of the lymphocyte population. Both D74- and RT-2-sensitized T cells constitutively secreted tumor necrosis factor-α, and both lymphocyte populations produced comparable amounts of the cytokine when co-cultured with either gliomacell line. Neither D74- and RT-2-sensitized effectors constitutively secreted γ-interferon (γ-IFN), but both populations produced γ-IFN when exposed to either glioma cell line in vitro. D74-sensitized T cells released significantly more γ-IFN than the RT-2 DLN lymphocytes. In vitro51Chromium-release assays indicated that RT-2-sensitized T cells were more cytotoxic for RT-2 targetsthan for the D74 line and that D74-sensitized effectors werealso more cytotoxic for RT-2 targets. To assess in vivo therapeutic efficacy, rats who had been inoculated intradermally with RT-2 cells three days earlier received an intravenous injection of RT-2- or D74-sensitized DLN cells (106Q cells/gram body weight) expanded after activation with Bryostatin-1 and ionomycin or an equal number of lymphokine-activated killer (LAK) cells. Tumordiameters were measured daily and revealed that injection of glioma- sensitized lymphocytes led to the elimination of tumor while treatmentwith LAK cells had no therapeutic benefit. These results indicate, that at least for these two glioma lines, γ-IFN release, rather than in vitro cytotoxicity, was a better predictor for in vivo immunotherapeutic efficacy of the glioma-sensitized, expanded T cells.