LAK cell proliferation by hollow fiber bioreactor

中空纤维生物反应器增殖LAK细胞

• 批准号: 04670902
• 负责人: ISHIDA Toshitake
• 金额: $ 1.41万
• 依托单位: OSAKA CITY UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04670902/
• 关键词: hollow fiber bioreactor; LAK cell

Osteosarcoma patients who had failed standard therapy and had multiple metastatic disease seems to be intractable. Rosenberg et al, have recently reported that combined therapy with LAK cells and IL-2 was capable of causing the regression of established metastatic cancer. In prior murine model, it is said that the therapeutic effect was directly related to the number of LAK cells and amount of IL-2 adminstered. In order to apply this adoptive immunotherapy to the advanced osteosarcoma patients, at first we have studied methods for in vitro expansion of LAK cells effective in immunotherapy.A hollow fiber biorcactor system (Cellmax 100) was used and its efficacy was investigated in LAK cell proliferatioin, PBL obtained from 20 ml of peripheral blood were cultured in an anti-CD 3 antibody coated flask for 7 days. About 2-4 *10 cells were harvested.These LAK cells were transfered to the hollow fiber bioreacter and cultured for further 2 weeks. The number of LAK cells did not increase in hollow fiber bioreactor. In their surface marker. Leu 4a (CD3+) cells were 98-99% at both 1 and 2 weeks. Leu 2a (CD8+) cells were 55% at 1 week, 74% at 2 weeks. Leu 3a (CD4+) cells were 48% at 1 week. 25% at 2 weeks. The ration of CD4/8 was decreased with time.We could not apply this adoptive immunotherapy to advanced osteosarcoma patients.

骨肉瘤患者标准治疗失败，并有多个转移性疾病似乎是难治性的。Rosenberg等最近报道了LAK细胞和IL-2的联合治疗能够使已建立的转移性癌症消退。在现有的小鼠模型中，据说治疗效果与LAK细胞的数量和IL-2的给药量直接相关。为了将这种过继免疫疗法应用于晚期骨肉瘤的治疗，我们首先研究了LAK细胞的体外扩增方法，并采用中空纤维生物反应器系统（Cellmax 100）对LAK细胞进行了体外扩增。收获2-4 × 10 ~ 6个细胞，将其转移至中空纤维生物反应器中继续培养2周。中空纤维生物反应器中LAK细胞数量无明显增加。在他们的表面标记。Leu 4a（CD 3+）细胞在1周和2周均为98-99%。Leu 2a（CD 8+）细胞在1周时为55%，2周时为74%。Leu 3a（CD 4+）细胞在1周时为48%。2周时为25%。CD 4/8比值随时间延长而下降，不能用于晚期骨肉瘤患者。