【摘要】 目的:评价细胞因子培养体系对经α-半乳糖酰基鞘氨醇(α-GalCer)体外扩增的NKT细胞的细胞因子生成的影响。方法:每份正常人外周血单个核细胞(MNC)标本分为3~4组:a[IL-2加IL-15,n=6]、b[IL-2加IL-15加IL-12加γ干扰素(IFN-γ),n=6]、c[IL-2加IL-15加IL-7,n=6]、d[IL-2加IL-15加IL-7加粒细胞集落刺激因子(G-CSF),n=4]。培养中加入α-GalCer和相应细胞因子,12d后用流式细胞计数法测定NKT细胞及其亚群数量,并用细胞胞内染色法测定细胞因子生成。结果:①扩增的NKT细胞CD4+亚群中IL-4+细胞比例高于CD4-亚群(P<0.01);2亚群中IFN-γ+细胞比例相当(P>0.05)。②各组NKT细胞中,b组IL-4+比例低于a组(P<0.01),c和d组均高于a组(均P<0.05),d和c组结果相当(P>0.05);而b组IFN-γ+比例略高于a组,但P>0.05,d组低于a组(P<0.05),c组与a组的结果相近(P>0.05)。③各组CD4+NKT细胞亚群中,b组IL-4+细胞比例低于a组(P<0.01),c和d组高于a组(均P<0.01),而c和d组之间差异无统计学意义(P>0.05);各组CD4-NKT细胞中,IL-4+细胞比例的差异无统计学意义(P>0.05);各组CD4+NKT细胞中,IFN-γ+细胞比例的差异无统计学意义(P>0.05);各组CD4-NKT细胞中,IFN-γ+细胞比例的差异无统计学意义(P>0.05)。结论:不同细胞因子组合的培养体系,能调节经α-GalCer扩增的Vα24NKT细胞IL-4和IFN-γ的生成,IL-7和G-CSF有助于IL-4的生成,而IL-12和IFN-γ有助于IFN-γ的生成。更多还原

【Abstract】 Objective:To investigate cytokine production by NKT cells following stimulation with α-galactosylceramide (α-GalCer) in vitro in the presence of different cytokine cocktails. Method:Mononuclear cells (MNCs) isolated from healthy adult peripheral blood were divided into group a(IL-2+IL-15, n=6),group b (IL-2+IL-15+IL-12+IFN-γ, n=6), group c (IL-2+IL-15+IL-7, n=6) and group d (IL-2+IL-15+IL-7+G-CSF, n=4), and were induced the expansion of Vα24 NKT cells with the stimulation of α-GalCer in the presence of different cytokine cocktails. On the 12th day, the numbers of NKT cell and its subtypes were measured, and the cytokine producing cells were especially identified via intercellular staining. Result:①Ratio of IL-4+ cells in CD4+ NKT cells was higher than in CD4-NKT cells (P<0.001), whereas ratio of IFN-γ+ cells was comparable in these two subsets. ②Ratio of IL-4+ NKT cells in group b (P=0.006) was lower than those in group c (P=0.016) or d (P=0.011) were higher than in group a. Ratio of IFN-γ+ NKT cells in group b(P=0.109) was higher and those in group d (P=0.019) was lower than in group a. ③The difference of ratio of IL-4+ NKT cells among four groups was observed in CD4+ subset but not in CD4-subset. The difference of ratio of IFN-γ+ NKT cells among four groups was not been when analyzing by means of CD4+ or CD4-subset. Conclusion:The significant influence of the cytokine mixture on the ability of Vα24 NKT cells to produce the cytokines was observed, as IL-7 and G-CSF were contributed to IL-4 production, IL-12 and IFN-γ were contributed to the IFN-γ production.更多还原