人外周血Vα24 NKT细胞和CD3~+CD56~+CIK细胞生物学特性的比较研究

目的进一步认识Vα24 NKT细胞和CIK细胞在生物学特性的差异。方法从人外周血单个核细胞扩增NKT细胞和CIK细胞;经免疫磁珠纯化获得高纯度的TCRVα24+NKT细胞和CD3+CD56+CIK细胞。运用流式细胞术测定纯化后的NKT细胞和CIK细胞的表型、细胞因子和细胞坏死相关因子的表达情况,并用DIOC18染色及流式细胞术测定纯化后的NKT细胞和CIK细胞的细胞杀伤活性。结果经纯化TCRVα24+Vβ11+NKT细胞和CD3+CD56+CIK细胞的纯度均达到>90%;纯化后的Vα24 NKT细胞多数为CD4+和DN NKT细胞,高表达TCRVα24、Vβ11、CD3、CD161,低表达CD56;而纯化后的CIK细胞则以CD8+T细胞为主,高表达CD3、CD56,低表达CD161,几乎不表达TCRVα24和Vβ11。在抗原刺激下,CD3+CD56+CIK细胞的IFN-γ、TNF-α、Perforin、FasL、TRAIL表达水平均高于NKT细胞,但CD3+CD56+CIK细胞几乎不分泌IL-4,而NKT细胞则分泌高水平的IL-4;CD3+CD56+CIK细胞对肿瘤细胞株K562、U937、Jurkat的杀伤率均远远高于NKT细胞。结论TCRVα24+NKT细胞和CD3+CD56+CIK细胞是两类完全不同的细胞群,在抗肿瘤免疫和免疫调节中可能起着不同的作用。

A comparative study on the biological characteristics of Vα24 NKT cells and CD3~+CD56~+CIK cells from human peripheral blood

Objective To confirm the difference in the biological characteristics between Vα24 natural killer T cells(NKT) and the cytokine-induced killer cell(CIK).Methods Vα24 NKT cells and CIK cells were expanded from human peripheral blood mononuclear cells.Purified TCRVα24~+ NKT cells and CD3~+CD56~+CIK cells were obtained by using Dynal beads.The phenotype and the levels of cytokine and necrosis-related factors expression on the purified NKT cells and CIK cells were determined by flow cytometry.The cytotoxicity of the purified NKT cells and CIK cells were measured by means of DIOC18 staining and flow cytometry.Results The proportions of TCRVα24~+Vβ11~+NKT cells and CD3~+CD56~+CIK cells were elevated up to 90% after purification.The majority of Vα24 NKT cells were CD4~+ and DN NKT cells.They demonstrated high levels of expression of TCRVα24,Vβ11,CD3 and CD161 and a low level of expression of CD56,but most purified CD3~+CD56~+CIK cells were CD8~+ T cells,with high levels of CD3 and CD56 expression,low level CD161 expression and little TCRVα24 and Vβ11expression.After antigen stimulation,the purified CD3~+CD56~+CIK cells showed higher levels of expression of IFN-γ,TNF-α,Perforin,FasL and TRAIL,compared to the NKT cells.CD3~+CD56~+CIK cells secreted little IL-4,whereas,Vα24~+NKT cells secreted high level of IL-4.In addition,the cytotoxic effect of the CD3~+CD56~+CIK cells on tumor cell lines K562,U937 and Jurkat were more intense than that of the NKT cells.Conclusion It is evident that TCRVα24~+ NKT cells and CD3~+CD56~+CIK cells differ absolutely in many ways and might play different roles in anti-tumor immunity and immune regulation.