LAK cell proliferation by hollow fiber bioreactor

骨肉瘤患者的标准治疗失败，并有多发性转移性疾病似乎是难治性的。Rosenberg等人最近报道，LAK细胞和IL-2联合治疗能够导致已建立的转移性癌症的消退。在先前的小鼠模型中，据说治疗效果与LAK细胞的数量和给药IL-2的量直接相关。为了将这种过继免疫疗法应用于晚期骨肉瘤患者，我们首先研究了体外扩增免疫治疗有效的LAK细胞的方法。采用中空纤维生物反应器体系（Cellmax 100），研究其对LAK细胞增殖的影响。取外周血20 ml PBL，在抗cd3抗体包被烧瓶中培养7 d。约收获2-4 *10个细胞。将这些LAK细胞转移到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细胞数量没有增加。在他们的表面标记。1周和2周时，Leu 4a （CD3+）细胞为98-99%。Leu 2a （CD8+）细胞1周时为55%，2周时为74%。1周时Leu 3a （CD4+）细胞为48%。2周25%。CD4/8的比例随着时间的延长而降低。我们不能将这种过继免疫疗法应用于晚期骨肉瘤患者。

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.