CD34^+造血祖细胞的聚集、CXCR3 mRNA的检测及CXCR3极化的研究

The aggregation of CD34^+ hematopoietic progenitors and the expression of CXCR3

目的 研究新鲜分离的和粒细胞单核细胞 集落刺激因子 (granulocytemacrophagecolonystimulatingfactor,GM CSF)刺激的CD34+ 造血祖细胞上CXC趋化性细胞因子受体 3(CXCchemokinereceptor 3 ,CXCR3)mRNA水平表达的区别 ,以及在γ 干扰素诱导蛋白 10 (IFN γinducibleprotein10 ,γIP 10 )和γ 干扰素诱导的单核因子 (monokineinducedbyIFN γ ,Mig)的作用下 ,GM CSF刺激的CD34+ 造血祖细胞的聚集作用和CXCR3分布的变化。方法 采用Northernblotting检测CXCR3mRNA水平的表达 ;2 4孔板细胞培养法观察细胞聚集作用 ;荧光标记抗体染色CXCR3后 ,以共聚焦免疫荧光显微镜观察CXCR3的极化现象。结果 GM CSF刺激的CD34+ 造血祖细胞上CXCR3mRNA表达水平明显高于新鲜分离的细胞 ;γIP 10和Mig能够诱导GM CSF刺激的CD34+ 造血祖细胞发生聚集现象和CXCR3重新分布现象 ,CXCR3聚集成束 ,并指向特定的方向。结论 GM CSF的刺激作用能够显著性提高CXCR3在CD34+ 造血祖细胞上的表达 ;γIP 10和Mig能够诱导GM CSF刺激的CD34+ 造血祖细胞的聚集作用和CXCR3的极化现象。

Objective To confirm the difference of CXCR3 mRNA expression on freshly isolated or GM CSF stimulated CD34 + hematopoietic progenitors, to investigate the aggregation of CD34 + hematopoietic progenitors and the redistribution of CXCR3 by the effect of γIP 10 and Mig. Methods Using Northern blot analysis to confirm the different expression of CXCR3 mRNA, using 24 well culture plates to assay the aggregation of CD34 + hematopoietic progenitors, using Polarization assay and immunofluroescence digital confocal microscopy to investigate the redistribution of CXCR3. Result Compared with the low level of CXCR3 mRNA in freshly isolated CD34+ progenitors, GM CSF stimulation can significantly up regulate the expression of CXCR3 mRNA in CD34+ progenitors. γIP 10 and Mig do not induce aggregation in freshly isolated CD34+ progenitors, but induce a strong aggregation in GM CSF stimulated cells. γIP 10 and Mig induce a rapid CXCR3 redistribution and the clusters of CXCR3 are lie in the leading edge of the cells and oriented towards a certain direction. Conclusion CXCR3 mRNA expression in CD34 + progenitors is up regulated by GM CSF. γIP 10 and Mig induce GM CSF stimulated CD34 + progenitor aggregation via CXCR3. Moreover, γIP 10 and Mig stimulate CXCR3 redistribution and cellular polarization in GM CSF stimulated CD34 + progenitors.