GM-CSF联合α-GalCer和IL-2促进人脐带血来源iNKT细胞的增殖

GM-CSF + α-GalCer + IL-2 promotes the proliferation of umbilical cord blood derived iNKT cells

目的观察GM-CSF联合α-GalCer和IL-2体外制备人脐带血来源自然杀伤T细胞(iNKT)的优势。方法采用GM-CSF联合α-GalCer和IL-2的方法体外制备iNKT细胞,与常规方法(加入α-GalCer联合IL-2)及只加IL-2进行对比,观察制备的单个核细胞的增殖情况,采用流式细胞仪检测单个核细胞中iNKT细胞、NK细胞、T细胞所占的比例,采用酶联免疫吸附试验(ELISA)测定培养上清中细胞因子IL-4和IFN-γ的浓度。结果与对照组(IL-2组)相比,GM-CSF+α-GalCer+IL-2组与α-GalCer+IL-2组均能促进iNKT细胞增殖,而前者iNKT细胞所占比例更高(P<0.05);此二组与对照组(IL-2组)相比IFN-γ的浓度增高更明显(P<0.05),但培养上清中IL-4的浓度增高不明显(P<0.05),GM-CSF+α-GalCer+IL-2组的高于α-GalCer+IL-2组。结论 GM-CSF联合α-GalCer和IL-2体外制备的iNKT细胞增殖能力较强,有望成为一种新的更好的iNKT细胞制备方法。

Objective To observe the effect of GM-CSF＋ α-GalCer ＋ IL-2 on the proliferation of invariant natural killer T cells（iNKT） from human umbilical cord blood. Methods Human umbilical cord blood mononuclear ceils were cultured in the presence of GM-CSF plus α -GalCer and IL-2, compared with conventional method （oL -GalCer plus IL-2） and only add IL-2. The iNKT cells, NK cells, T cells were count by flow cytometry, and the concentrations of IL-4 and IFN-γ in serum were detected by enzyme-linked immunosorbent assay（ELISA）. Results Both of GM-CSF＋ α -GalCer＋ IL-2 and α -GalCer ＋ IL-2 promoted iNKT cells proliferation, with much more in former than in latter. The concentration of IFN -γ in culture supernatants was higher in GM-CSF＋ α -GalCer＋ IL-2 group than in α -GalCer ＋ IL-2 group, but with no difference for IL-4 between GM-CSF＋ α-GalCer＋ IL-2 group and α-GalCer ＋ IL-2 group. Conclusion GM- CSF＋ α -GalCer＋ IL-2 promoted mostly the proliferation of iNKT cells, would be a new better protocol for preparation of iNKT cells.