Large-scale production of natural cytokines during activation and expansion of human T lymphocytes in hollow fiber bioreactor cultures
Large-scale production of natural cytokines during activation and expansion of human T lymphocytes in hollow fiber bioreactor cultures
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DOI：
10.1097/00002371-199907000-00003
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发表时间：
1999-07-01
影响因子：
3.9
通讯作者：
Bast, EJEG
中科院分区：
文献类型：
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作者：
Lamers, CHJ;Gratama, JW;Bast, EJEG
我们研究了中空纤维生物反应器培养系统中人T淋巴细胞活化和扩增过程中多种天然细胞因子的大规模生产。使用植物血凝素加重组白细胞介素 2 (IL-2) 激活外周血单核细胞 (PBMC)。植物血凝素要么在整个 15-16 天培养期间存在于中空纤维生物反应器中，要么仅在培养袋中 PBMC 预激活的 20 小时期间存在。扩增的T淋巴细胞主要为CD3(+)、8(+)，并在培养第9天至第13天之间发挥最大的天然、活化、双特异性单克隆抗体重定向和凝集素依赖性细胞溶解活性。 IL-1 和 IL-1 仅产生少量。 IL-8 和淋巴毒素主要在培养第一周产生。在细胞因子浓度达到峰值时收获中空纤维生物反应器培养物上清液，每输入 10(8) 个 PBMC 将产生 3.7 至 4.9 μg 的 IL-8（第 2 或 3 天）和 0.02 至 0.5 μg 的淋巴毒素（第 6 或 7 天）。肿瘤坏死因子-α 和 IL-6 在 15 或 16 天的整个培养期间产生：每 10(8) 个 PBMC 输入。 0.1至0.4μg肿瘤坏死因子-α（第2或3天）和0.03至0.5μg IL-6（第15或16天）。干扰素-γ和粒细胞-巨噬细胞集落刺激因子的产生从培养开始开始，在15天或16天培养期结束时达到峰值水平，此时每10(8)产生2.1至17.7μg/ml干扰素-γ和0.4至4.2μg粒细胞-巨噬细胞集落刺激因子。 PBMC 输入。肿瘤坏死因子-α、IL-6、干扰素-γ和粒细胞-巨噬细胞集落刺激因子的产生与淋巴细胞增殖的程度成正比。这些结果证明了中空纤维生物反应器培养物在主要CD3(+),8(+) T淋巴细胞的激活和扩增过程中产生天然细胞因子的有用性。
We studied the large-scale production of a variety of natural cytokines during the activation and expansion of human T lymphocytes in a hollow fiber bioreactor culture system. Peripheral blood mononuclear cells (PBMC) were activated using phytohemagglutinin plus recombinant interleukin-2 (IL-2). Phytohemagglutinin was either present in the hollow fiber bioreactor during the entire 15-16-day culture period or only during the 20-h preactivation of the PBMC in culture bags. The expanding T lymphocytes were mainly CD3(+),8(+) and exerted maximal natural, activated, bispecific monoclonal antibody-redirected and lectin-dependent cytolytic activities between days 9 and 13 of culture. IL-1 and IL-1 were only produced in low amounts. IL-8 and lymphotoxin were primarily produced during the first week of culture. Harvest of the hollow fiber bioreactor culture supernatant at the time of peak cytokine concentration would have yielded per 10(8) PBMC input between 3.7 and 4.9 mu g of IL-8 (at days 2 or 3), and between 0.02 and 0.5 mu g of lymphotoxin (at days 6 or 7). Tumor necrosis factor-alpha and IL-6 were produced during the entire culture period of 15 or 16 days: per 10(8) PBMC input. between 0.1 and 0.4 mu g of tumor necrosis factor-alpha (at days 2 or 3) and between 0.03 and 0.5 mu g of IL-6 (at days 15 or 16). Production of interferon-gamma and granulocyte-macrophage colony-stimulating factor started from initiation of cultures onwards to reach peak levels at the end of the 15- or 16-day culture period, yielding at that time between 2.1 and 17.7 mu g/ml of interferon-gamma and between 0.4 and 4.2 mu g of granulocyte-macrophage colony-stimulating factor per 10(8) PBMC input. The production of tumor necrosis factor-alpha, IL-6, interferon-gamma, and granulocyte-macrophage colony-stimulating factor was proportional to the extent of lymphocyte multiplication. These results demonstrate the usefulness of hollow fiber bioreactor cultures to produce natural cytokines during the activation and expansion of predominantly CD3(+),8(+) T lymphocytes.