小鼠外周血嗜碱性粒细胞活化后的CD200R表达

CD200R Expression in Activated Peripheral Blood Basophiles of Mice

目的探讨流式细胞术鉴定小鼠外周血嗜碱性粒细胞方法,并分析其活化后表面分子CD200R的表达。方法分离BALB/c小鼠外周血,以抗-IgE-FITC和抗-CD4-PE/cy7与抗-CD45R B220-PE/cy7双色免疫荧光染色结合流式细胞术分析,鉴定小鼠外周血嗜碱性粒细胞;正常以及卵清蛋白(OVA)致敏的BALB/c小鼠外周血,抗-IgE刺激后抗-IgE-FITC、抗-CD4-PE/cy7、抗-CD45R B220-PE/cy7和抗-CD200R-PE三色免疫荧光染色结合流式细胞术检测CD200R的表达。结果小鼠外周血中嗜碱性粒细胞的数量占白细胞总数为0.7%,抗-IgE刺激后嗜碱性粒细胞中CD200R+表达率由3.62%升至31.6%(P<0.01)。OVA致敏小鼠外周血,经O-VA体外刺激后嗜碱性粒细胞CD200R+表达率由13.43%升至23.5%(P<0.05)。结论 IgE+CD4-CD45R-双色荧光标记可以鉴定小鼠嗜碱性粒细胞;CD200R可以作为小鼠嗜碱性粒细胞重要的表面活化分子。

Objective To establish the method of flow cytometry in identifying mouse peripheral blood basophiles and to investigate the expression of the surface molecule CD200R after the activation of basophiles. Methods BALB/c mouse peripheral blood was separated. The method of flow cytometry combined with immunofluorescence staining with antiIgEFITC, antiCD4PE/ey7 and antiCD45RPE/cy7 was used to identify mouse peripheral blood basophiles. The peripheral blood of normal BALB/c mice and allergic mice induced by ovalbumin（OVA） was incubated with anti- IgE at 37 ℃ for 30 min, then the method of flow cytometry combined with immunofluorescence staining with anti IgEFITC, antiCD4PE/cy7, antiCD45RPE/cy7, antiCD200RPE was used to detect the expression of the surface molecule CD200R of basophiles. Results IgE＋CD4-CD45Reells accounted for 0.7 % of all circulating leuco- cytes. After stimulating with anti-IgE, the expression rate of CD200R＋ was up to 31.6 % from 3.62 %（P 〈 0.01 ）. In the peripheral blood of allergic mice induced by OVA, the expression rate of CD200R＋ was up to 23.5 % from 13.4 % （P 〈 0.05）. Conclusion IgE＋CD4CD45R twocolor fluorescence labeling can be used to identify the mouse periph eral blood basophiles. CD200R can be a surface molecule to mirror the activation of the mouse peripheral blood ba sophiles.