人外周血树突状细胞诱导及免疫学特性研究

目的]从人外周血分离、纯化、扩增树突状细胞(DC),并对其形态学和免疫学特性进行初步探讨.方法]从人外周血分离DC前体细胞(主要为CD14 细胞)用重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)和重组人白细胞介素-4(rhIL-4)联合培养,诱导扩增成熟DC.观察DC形态、分析DC表型、核型及检测DC激发同种异体淋巴细胞增殖能力.结果]分离的DC前体经rhGM-CSF和rhIL-4共同培养1周后,可获得大量成熟DC,扩增了24.5倍,纯度达90%以上.DC高表达分化抗原CD86、CD40、HLA-DR、CD83、CDIa,能强烈激活同种异体T淋巴细胞增殖.结论]人外周血CD14 细胞经体外诱导培养,可以生成大量功能成熟的DC,从而为进一步开展DC的基础研究和临床应用打下基础.

An Experimental Study on Inducement and Immunological Features of Dendritic Cells from Human Peripheral Blood

To amplify dendritic cells (DC) from human peripheral blood and assay their morphological and immumlogical features.The mononuclear cells isolated from human peripheral blood were cultured with the supplement of rhGM-CSF and rhIL-4 to generate DC.The morphological characteristics of those cells were observed and the phenotype features were analysed.The nucleio-type was analysed and the proliferation of allogeneic T lymphocytes stimulated by DC was detected also.Monocytes from human peripheral blood were cultured with rhGM-CSF and rhIL-4 for one week, and then a large number of maturative DC with high puring(90%) were generated,which highly expressed differential antigens CD86, CD40, HLA-DR, CDIa, CD86 and CD83. Meanwhile, the induced DC can effectively stimulate proliferation of allogeneic T lymphocytes.Conclusion]A large number of maturative DC were induced from human peripheral blood, and may provide foundation for further studies on DC and their clinical applications.