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目的 研究GM CSF诱导人脐血CD34 + 造血干细胞上CXCR3的表达。方法 采用流式细胞仪 ,实时定量逆转录PCR(RT PCR)分析 ,及其配体γIP 10和Mig诱导的趋化和粘附作用分析。结果 CXCR3也表达在受GM CSF刺激后的CD34 + 造血干细胞上 ,但它在新鲜分离的CD34 + 造血干细胞上不表达。应用实时定量逆转录PCR技术检测新鲜分离的CD34 + 造血干细胞有较低水平的CXCR3mRNA表达 ,而GM CSF能上调CXCR3的表达。用抗CXCR3单克隆抗体 (mAb)能阻断γIP 10和Mig诱导的造血干细胞趋化作用 ,证实γIP 10和Mig是通过CXCR3而发挥作用的。γIP 10和Mig通过CXCR3不仅可诱导趋化作用而且还可诱导GM CSF刺激后的CD34 + 造血干细胞粘附和聚集作用 ,而抗CXCR3mAb能阻断γIP 10和Mig这些功能 ,但不能阻断SDF 1α的作用。γIP 10和Mig能提高整合素(CD49a和CD49b)的表达 ,这在GM CSF刺激后CD34 + 造血干细胞的粘附作用中发挥重要作用。结论 在细胞因子 /趋化因子微环境中 ,CXCR3 γIP 10和 Mig受体配体复合物及GM CSF对CD34 + 造血干细胞分化成淋巴干细胞和髓系干细胞以及后来的免疫 /炎症细胞的生理和病理过程起特别重要作用。这些过程包括CD34 + 造血干细胞的迁移、再定位、分化和成熟。
Objective To investigate the expression of CXCR3 on human umbilical cord blood CD34 + hematopoietic stem cells induced by GM CSF. Methods Flow cytometry, real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis and chemotaxis and adhesion induced by its ligands γIP 10 and Mig were analyzed. Results CXCR3 was also expressed on CD34 + hematopoietic stem cells stimulated with GM CSF, but it was not expressed on freshly isolated CD34 + hematopoietic stem cells. Real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to detect CXCR3 mRNA expression in freshly isolated CD34 + hematopoietic stem cells. GM CSF up-regulated the expression of CXCR3. Anti-CXCR3 monoclonal antibodies (mAb) blocked the chemotaxis of γIP 10 and Mig-induced hematopoietic stem cells, demonstrating that γIP 10 and Mig play a role through CXCR3. γIP 10 and Mig through CXCR3 not only induced chemotaxis but also induced GM CSF-stimulated CD34 + hematopoietic stem cell adhesion and aggregation, whereas anti-CXCR3 mAb blocked the function of γIP 10 and Mig but failed to block SDF 1α Role. γIP 10 and Mig increased the expression of integrins (CD49a and CD49b), which play an important role in the adhesion of CD34 + hematopoietic stem cells after GM CSF stimulation. Conclusions In the cytokine / chemokine microenvironment, the CXCR3 γIP 10 and Mig receptor ligand complexes and GM CSF differentiate CD34 + hematopoietic stem cells into lymphocyte and myeloid stem cells and subsequent immune / inflammatory cell physiology and pathology The process plays a particularly important role. These processes include the migration, repositioning, differentiation and maturation of CD34 + hematopoietic stem cells.