树突状细胞前体细胞亚群四色荧光分析方法探讨

目的建立树突状细胞(DC)前体细胞(pDC)亚群四色荧光检测方法,探讨以Lin-人类白细胞抗原(HLA)-DR+细胞群和以CD34-Lin-HLA-DR+细胞群设门检测不同来源标本pDC亚群的适用性。方法以Lin-HLA-DR+细胞群和以CD34-Lin-HLA-DR+细胞群设门分别检测健康成人外周血、脐带血、骨髓、粒细胞集落刺激因子(G-CSF)动员外周血pDC亚群各15例,比较两种设门方法的pDC检测结果。结果以该两种方法设门,pDC1/pDC2比值:外周血=脐带血>骨髓>G-CSF动员外周血;对同一种标本,两种设门方法之间比较,pDC1/pDC2比值差异无统计学意义;外周血CD34-Lin-HLA-DR+/单个核细胞(MNC)与Lin-HLA-DR+/MNC比值、pDC/CD34-Lin-HLA-DR+比值与pDC/Lin-HLA-DR+比值差异无统计学意义;而脐带血、骨髓、G-CSF动员外周血的CD34-Lin-HLA-DR+/MNC比值均显著性低于Lin-HLA-DR+/MNC比值,pDC/CD34-Lin-HLA-DR+比值显著性高于pDC/Lin-HLA-DR+比值。结论检测pDC亚群,对一般外周血标本,以Lin-HLA-DR+细胞群设门或以CD34-Lin-HLA-DR+细胞群设门,均可取得理想结果;对CD34+细胞含量相对丰富的脐带血、骨髓、G-CSF动员外周血等的标本,以CD34-Lin-HLA-DR+细胞群设门能明显优化检测结果。

Analysis of predendritic cell subsets in cord blood, peripheral blood and bone marrow

Objective To explore a suitable method to detect predendritic cell(pDC) subsets in cord blood,peripheral blood,bone marrow and G-CSF-mobilized blood.Methods pDC subsets in cord blood,peripheral blood,bone marrow and G-CSF-mobilized blood were detected gated by Lin~-HLA-DR~ cell population or CD34~-Lin~-HLA-DR~ cell population.pDC1/pDC2,CD34~-Lin~-HLA-DR~ cell/mononuclear cell(MNC) and Lin~-HLADR~ cell/MNC,pDC/CD34~-Lin~-HLA-DR~ cell and pDC/Lin~-HLA-DR~ cell were compared between two gating methods.Results pDC1/pDC2 ratio in cord blood,peripheral blood,bone marrow and G-CSF-mobilized blood, CD34~-Lin~-HLA-DR~ cell/MNC ratio and Lin~-HLA-DR~ cell/MNC ratio,pDC/CD34~-Lin~-HLA-DR~ cell ratio and pDC/Lin~-HLA-DR cell ratio in peripheral blood were not remarkably distinct between two gating methods(P>0.05),but detecting CD34~-Lin~-HLA-DR~ cell/MNC were lower than Lin~-HLA-DR~ cell/MNC(P<0.05),pDC/CD34~-Lina-HLA-DR~ cell were higher than pDC/Lin~-HLA-DR~ cell in cord blood,bone marrow and G-CSF-mobilized blood(P<0.05).Conclusion pDC subsets in cord blood,bone marrow and G-CSF mobilized blood should be analyzed by gating by CD34~-Lin~-HLA-DR~ cell population,and pDC subsets in peripheral blood can be analyzed by gating by Lin~-HLA-DR~ cell population or CD34~-Lin~-HLA-DR~ cell population because enriching CD34~ cells gated by Lin~-HLA-DR~ cell population were very fewer than in cord blood,bone marrow and G-CSF-mobilized blood.