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目的观察不同类型、不同浓度的抗CD3单抗对细胞因子诱导的杀伤细胞(CIK细胞)体外增殖能力及其抗肿瘤特性的影响,优化扩增方案。方法取多名健康供血者血液用淋巴细胞分离液分离外周血单个核细胞,加入不同类型(OKT3、UCHT1、HIT3a)、不同浓度、不同组合的抗CD3单抗及细胞因子(γ-IFN、rhIL-22),诱导生成CIK细胞,体外培养12 d。观察CIK细胞的扩增情况,采用MTT法测定其杀瘤活性。结果 (1)等量混合3种不同类型的抗CD3单抗,CIK细胞增殖反应优于添加单一类型抗CD3单抗(P<0.05)。(2)抗CD3单抗浓度增加,CIK细胞的增殖效应及细胞毒活性反而下降。(3)不同rhIL-2浓度对CIK细胞的增殖效应及细胞毒活性均无明显影响。结论培养CIK细胞时混合不同类型的抗CD3单抗有利于CIK细胞的体外扩增。
Objective To observe the effects of different types and concentrations of anti-CD3 monoclonal antibodies on in vitro proliferation and anti-tumor properties of cytokine-induced killer cells (CIKs), and to optimize the amplification scheme. Methods Peripheral blood mononuclear cells (PBMCs) were isolated from lymphocytes using multiple blood donors. Anti-CD3 monoclonal antibodies (mAbs) and cytokines (γ-IFN, rhIL -22), induce the generation of CIK cells cultured in vitro 12 d. The proliferation of CIK cells was observed and the cytotoxic activity was measured by MTT assay. Results (1) Three different types of anti-CD3 McAbs were mixed equally, CIK cells proliferated better than single anti-CD3 monoclonal antibody (P <0.05). (2) The anti-CD3 monoclonal antibody concentration increased, CIK cell proliferation and cytotoxic activity but decreased. (3) Different concentrations of rhIL-2 had no significant effect on proliferation and cytotoxicity of CIK cells. Conclusion Mixing different types of anti-CD3 mAb in CIK cells is beneficial to in vitro expansion of CIK cells.