L-精氨酸对树突状细胞表型及功能的影响

Effect of L arginine on phenotype and function of dendritic cells

目的体外分离培养人外周血树突状细胞（DCs），并探讨L-精氨酸（L-arg）对DCs表型表达及抗原提呈能力的影响。方法对照组密度梯度离心法分离人外周血单个核细胞，取贴壁细胞加入粒-巨噬细胞集落刺激因子（GM-CSF）、白细胞介素-4（IL-4），第6天加入肿瘤坏死因子一仪（TNF-α）刺激DCs成熟；实验组在此基础上加入不同剂量L-arg继续培养；第8天用倒置显微镜观察细胞形态，流式细胞仪检测细胞表型，混合淋巴细胞反应观察各组细胞的抗原提呈能力，在酶标仪490nm处测出光密度（OD）值。结果对照组和实验组均表现出DCs典型形态学特征，实验组CD80、CD83、CDld、HLA-DR表达增加，混合淋巴细胞反应中实验组反映T淋巴细胞增殖效应强度的OD值高于对照组。结论L-arg可促进DCs成熟，增加DCs表型的表达，并提高DCs的抗原提呈能力。

Objective To culture and isolate dendritic cells （DCs） from human peripheral blood in vitro and to explore the effect of L arginine （ L arg） on the phenotype and antigen presenting ability of DCs. Methods The mononuclear cells were separated from human peripheral blood by density gradient eentrifugation in control group. The adherent ceils were cultured with human recombinant granulocyte macrophage colony stimulating factor （ GM CSF） and human recombinant interleukin 4 （IL 4） , and then tumor necrosis factor-α （TNF -α） was added on the 6th day. On this basis, the experimental groups were cuhured with different doses of Larg. On the 8th day, the change in morphology was observed with inverted microscope, meanwhile antigen presenting ability was reflected from mixed lymphocyte reaction, and phenotypic characterization was detected by flow cytometer. Results All the groups showed a characteristic dendritic morphology, however, DCs expressed higher level of CD80, CD83, CDlα and HLA DR in experimental groups. In addition, a higher optical density value was demonstrated by experimental groups DCs in mixed lymphocyte reactions. Conclusion Larginine can promote the mature of DCs, and increase the expression of phenotype and antigen presenting ability of DCs.