对过继免疫治疗中外周血单个核细胞采集术的探讨

Investigation of cytapheresis of peripheral blood mononuclear cells during adoptive immunotherapy

目的:探讨在恶性肿瘤患者过继免疫治疗中外周血单个核细胞的采集技术。方法:根据患者外周血单个核细胞计数值,应用CS-3000PLUS血细胞分离机,采集外周血单个核细胞,运用Ficoll法分离纯化,再用贴壁法分离出贴壁细胞进行诱导分化为树突状细胞(DC),未贴壁细胞(主要为淋巴细胞)诱导分化为肿瘤杀伤细胞(CIK)或体外扩增为记忆性淋巴细胞(ALT),最后进行细胞回输。结果:在8例13人次的采集中,处理总循环血量,获得单个核细胞总数,单个核细胞纯度,细胞采集效率,血小板丢失率分别为(6.99±1.74)L,(3.47±1.44)×109/L,(90±6)%、(51.3±13)%、(43.5±13.1)%。DC加CIK治疗1例,CK治疗3例,化疗后ALT免疫重建治疗4例,均取得了一定的临床疗效。结论:外周血单个核细胞采集术,不良反应发生率低、单个核细胞的采集效率高,用于恶性肿瘤过继免疫治疗的临床效果较好。但采集后血小板的丢失严重,对于恶性肿瘤化疗后血小板减低的患者应引起关注。

Objective:To explore the collection method of peripheral blood mononuclear cells on malignant tumor patients during adoptive immunotherapy.Method:According to the count of patients' peripheral blood mononuclear cells,peripheral blood mononuclear cells were collected by blood cell separator CS-3000PLUS and purified by Ficoll method.Adherent cells were separated out of them and differentiated to dendritic cells(DC),while unadherent cells(lymph cells mainly) were separated and differentiated to cytokine-induced killer cells(CIK) or amplified to memory lymphocytes(ALT) in vitro.They were all infused back into patients.Result:In 8 collections of 13 person-times,the whole volume of disposal circulation,the count of harvest mononuclear cells,purity of mononuclear cells,cell collection efficiency and platelet loss rate were(6.99±1.74) L,(3.47±1.44)×109/L,(90±6)%,(51.3±13)% and(43.5±13.1)%,respectively.1 case of DC and CIK therapy,3 cases of CK,4 of ALT immunity reconstruction after chemotherapy were operated and all had reached certain clinical effect.Conclusion:The cytapheresis of peripheral blood mononuclear cells has low adverse reaction incidence,high cell collection efficiency and good clinical effect on malignant tumor patients during adoptive immunotherapy.However,due to its high loss probability of platelet,we should pay attention to those platelet-reduced malignant tumor patients after chemotherapy.