猪红细胞CR1-like膜分布状态的初步研究

Initial study of CR1-like expressed on porcine erythrocytes surface

为进一步研究猪红细胞的天然免疫调控机制,本研究合成免疫抗原肽段,制备获得了猪CR1-like单克隆抗体,并应用CR1-like单克隆抗体通过荧光免疫细胞化学技术观察猪红细胞CR1-like的天然分布状态,运用流式细胞检测技术检测了膜流动性变化对猪红细胞CR1-like发挥免疫黏附功能的影响。结果显示,于荧光显微镜下观察,猪红细胞表面可见绿色荧光,且经膜固定处理的猪红细胞表面的绿色荧光分布呈片状,未经膜固定处理的猪红细胞表面的荧光点呈颗粒状分布,较膜固定组分布明显成簇化;流式细胞仪检测荧光强度发现,膜固定组、未固定组均明显高于空白对照组,差异极显著(P<0.01);同时未固定组平均荧光强度明显高于膜固定组,差异极显著(P<0.01)。证实天然状态下猪红细胞表面的CR1-like分布呈分散状态,发挥免疫功能时,CR1-like表现出多价高效的结合特性,其分布状态表现为颗粒样成簇分布。

In order to study the molecular mechanisms of porcine erythrocyte CR1-like,monoclonal antibody against porcine CR1-like was produced.The distribution of CR1-like in porcine erythrocytes was observed by fluorescence immunocytochemistry.Effects of erythrocyte membrane fluidity on the immune adhere function of porcine CR1-like was observed in vitro by flow cytometric method.The green fluorescence dots were scattered around the glutaraldehyde（GA）-fixed erythrocytes surface,but clustered on unfixed erythrocytes.The mean fluorescence intensity（MIF）of the GA-fixed and unfixed erythrocytes were all significantly higher than that of the control erythrocytes（P〈0.01）,the MIF of the GA-fixed erythrocytes was lower than the unfixed erythrocytes（P〈0.01）.CR1-like express scattered around porcine erythrocytes surface,but clustered when the erythrocytes adhere the opsonized immune complex.