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目的:探索体外培养的干扰素-树突状细胞(IFN-DC)和白介素4-树突状细胞(IL-4-DC)在表型及功能上的区别。方法:人外周血单核细胞在体外分别用巨噬细胞集落刺激因子(GM-CSF)联合IFN-α或者GM-CSF联合IL-4诱导培养为IFN-DC或IL-4-DC,随后加入TNF-α促进DCs成熟,流式细胞仪检测两种DC的表型;用负载CMVpp65蛋白的两种DCs刺激自体T淋巴细胞激活,细胞内染色方法检测分泌IFN-γ的T细胞的比例。结果:IFN-DC和IL-4-DC均具有典型的树突状细胞形态和免疫表型,但IFN-DC表达更高水平的CD80、CD83、CD86。负载CMV pp65蛋白的不成熟IFN-DC及不成熟IL-4-DC能够分别诱导(0.28±0.07)%及(0.12±0.02)%的自体CD8+T细胞激活,两者差异有统计学意义(P<0.05);成熟IFN-DC及成熟IL-4-DC能够分别激活(1.03±0.21)%及(0.35±0.14)%的自体CD8+T细胞激活,两者差异有统计学意义(P<0.05)。结论:与IL-4-DC相比,IFN-DC表达更高水平的CD80、CD83、CD86,IFN-DC负载CMV pp65蛋白能够更有效激活人抗原特异性CD8+T细胞。
Objective: To explore the difference in phenotype and function of interferon - dendritic cells (IFN - DCs) and interleukin - 4 (DCs) - dendritic cells (IL - 4 - DCs) cultured in vitro. Methods: Human peripheral blood mononuclear cells were cultured in vitro as IFN-α or IL-4-DC with GM-CSF plus IFN-α or GM-CSF combined with IL-4, respectively, TNF-α promoted the maturation of DCs. The phenotypes of the two DCs were detected by flow cytometry. Autologous T lymphocyte activation was stimulated by two kinds of DCs loaded with CMVpp65 protein. The proportion of T cells secreting IFN-γ was detected by intracellular staining. Results: Both DCs and IL-4-DCs displayed typical dendritic cell morphology and immunophenotype, but IFN-DCs expressed higher levels of CD80, CD83 and CD86. Immature IFN-DCs and immature IL-4-DCs loaded with CMV pp65 protein were able to induce (0.28 ± 0.07)% and (0.12 ± 0.02)% autologous CD8 + T cell activation, respectively P <0.05). Mature IFN-DC and mature IL-4-DC could activate (1.03 ± 0.21)% and (0.35 ± 0.14)% of autologous CD8 + T cell activation respectively, with statistical significance (P < 0.05). CONCLUSION: Compared with IL-4-DC, IFN-DC expresses higher levels of CD80, CD83 and CD86, and IFN-DC can effectively activate human antigen-specific CD8 + T cells by loading CMV pp65 protein.