Identification Of Immune Adherence Receptor On Porcine Erythrocyte Membrane

Objective:To identify and detect the immune adherence receptor CRl-like of porcine erythrocyte,and estimate the molecular size and the quantity of disulfide bonds of CRl-Like.Method:The method of indirect immune fluorescence was used to detect CR1 of porcine erythrocyte.Green fluorescent protein Escherichia coli(GFP-E.coli) were treated with 20% fresh rabbit serum or EDTA serum, which as ICs co-incubated with fresh erythrocytes or anti-CRl-like mAb preincubating erythrocytes of porcine,then observed under the fluorescence microscopy; the Latex particle complex also treated with 20% fresh rabbit serum or EDTA serum and co-incubated with fresh erythrocytes or anti-CRl-like mAb preincubating erythrocytes of porcine, then also observed on fluorescence microscopy. IP experiment was used to purified the aim protein and the molecular size and the quantity of disulfide bonds of aim protein was estimated by SDS-PAGE and WB.Results:Under the fluorescence microscopy, we observed that fresh rabbit serum complement-opsonized GFP-E. coli adhered to the erythrocytes,EDTA serum opsonized GFP-E.coli dosen’t adhere to the erythrocytes, the erythrocytes preincubated by anti-CRl-like mAb have no GFP-E.coli and the erythrocytes preincubated by isotype antibody adhered GFP-E.coli as the first group;the result of Latex particle complex were consistent with GFP-E.coli.In non-reducing condition, the aim protein eluted by IP have two protein 135KD and 200KD.In reducing condition, the aim protein splited three protein 70KD 50KD 15KD. In WB experiment,two splited protein 50KD and 15KD could bind anti-CRl-like mAb.Conclusion:The immune adherence receptor CRl-like existence on erythrocyte of porcine.CRl-like have two form protein 135KD and 200KD. In reducing condition, CRl-like splited three protein 70KD 50KD 15KD,and exist two or three disulfide bonds in CRl-like inside.