Studies On Molecular Mechanism Of Porcine Erythrocytes Presenting Immune Complexes

Objective:This thesis aimed at researching the function of porcine erythrocyte presenting immune complexes. In order to provide theoretical basis for the molecular mechanism of animal erythrocyte immune function in depth.Method:In this study,we regarded the Changbai pig as research object,separating PAM and porcine erythrocytes routinely,regarding FITC-WT E.coli as report factor, which built an in vitro model of porcine erythrocytes clearing immune complexes; Observing and analyzing the fluorescent E.coli adhering to porcine erythrocytes and transfer to PAM from porcine erythrocyte by using Laser Scanning Confocal Microscopy(LSCM) and Flow Cytometry(FCM); Analyzing the porcine CRl-like molecular mechanism during the porcine erythrocytes clearing immune complexes with FCM.Results:The separated PAM after recovery culture was observed by inverted microscope,cells were round shape,uniform size,good activity,showed half adherence. After trypan blue staining,PAM survival rate was 97.2% counted by microscopy; when add ink to the system of recovery training PAM,the inverted microscope microscopic examination showed cell with phagocytic particles and PAM phagocytosis rate was 98.8%; Upright microscope observation showed that the FITC-WT E.coli was a short rod like morphology,size of 1-2 m; Under fluorescence field,WT E.coli showed strong green fluorescence and the fluorescence labeling rate was 92.3%; Observation of porcine erythrocytes,PAM,FITC-WT E.coli incubated system under LSCM showed that the fluorescence intensity of FITC-WT E.coli decreased gradually until disappear between six time points(2h5min,2hllmin,2h23min,2h27min,2h36min,2h59min); FCM results showed that after incubation with porcine erythrocytes adhered with FITC-WT E.coli,the mean fluorescence intensity of PAM are extremely higher than usual; Analyzing by FCM,when incubated the porcine erythrocytes which were adhered with WT E.coli with PAM and then reacted with atibodies after recycling,thier mean fluorescence intensity were evidently higher than common,but there is no obvious difference to the porcine erythrocytes only incubated with PAM.Conclusion:The study had found that PAM could competitively bond the opsonized pathogens adhering to the porcine erythrocytes membrane and then phagocytosed them,which made for the clearance of immune complexes or pathogens; In this process,the physical form of erythrocytes was not destroyed,but the number level of its main function protein CR1-like reduced.