猪外周血单核细胞来源的树突状细胞体外诱导培养

Porcine peripheral blood monocyte-derived dendritic cells cultured in vitro

为建立猪外周血单核细胞来源的树突状细胞(Mo DC)体外培养模型,从猪外周新鲜血液中分离外周血单核细胞(PBMC),通过贴壁法获得树突状细胞前体细胞,采用重组猪的集落共刺激因子(rp GM-CSF)和白细胞介素4(rp IL-4)双因子诱导及脂多糖(LPS)刺激成熟法,收集不同时间段的细胞,利用扫描电子显微镜观察其形态,流式细胞分析仪检测表面分子表达率及其对FITC-dextran的吞噬能力,混合淋巴细胞反应检测细胞对同种异体T细胞的刺激能力。结果表明,经体外诱导的细胞具有典型的树突状形态;经脂多糖刺激的树突状细胞表型分子CD1a、CD80、CD86、SLAII、CD172a与未经刺激的树突状细胞有明显增高,其吞噬能力有所下降,刺激同种异体T淋巴细胞的能力增强。本研究成功建立了猪Mo DC的体外诱导培养方法,为进一步研究其在机体免疫调节和抗病毒感染中的作用奠定了基础。

To develop a method for maturation induction of dendritic cells（ DCs） in vitro,porcine peripheral blood monocyte-derived DCs（ Mo DCs） were induced in the presence of rp GM-CSF,rp IL4 and lipopolysaccharide（ LPS）. The cells were harvested to detect the morphological,phenotypic and functional characteristics. FITC-dextran endocytic activity and the surface molecules of Mo DCs were measured with flow cytometer,and T lymphocyte proliferation following Mo DCs stimulation were detected by CCK-8 assay. The induced Mo DCs showed typical dendritic morphology, and the expressions of the surface molecules CD1 a,CD80,CD86,SLAII,and CD172 a were increased by LPS treatment from53. 8%,42. 4%,23. 4%,54. 2%,48. 2% to 55. 2%,53. 6%,43. 6%,65. 4%,61. 1%,respectively. The phagocytic ability of non-LPS-treated DC to uptake FITC-dextran was higher than that of LPS-treated DC. The proliferation of allogenetic T lymphocyte was enhanced in LPS-treated DCs. In conclusion,the porcine Mo DC was successfully induced and cultured in vitro,laying a foundation for further study on the role of porcine Mo DC in immune regulation and anti-virus infection.