Effects Of PRRSV Infection On Autophagy And Transcription Level Of Antigen Presenting Genes In Dendritic Cells

The pathogenic mechanism of PRRSV is not yet clear,so it is currently unable to eradicate the disease effectively.Recent studies have shown that PRRSV induces autophagy after infection of the body,and autophagy of the body cells is used to replicate the virus itself,and the autophagic target Protein m TOR is a serine/threonine kinase that is an important target in the autophagy signaling pathway and can regulate phosphorylation of downstream proteins,thereby affecting the normal operation of downstream pathways.Dendritic cells are natural bridges that link natural immunity and specificity and play an important role in immune responses.Therefore,the purpose of this dissertation is to study the autophagy caused by PRRSV infection in dendritic cells,and then use autophagy activators and inhibitors to regulate the main targets of autophagy,so as to prevent PRRSV replication by regulating autophagy levels.The experiment was divided into 6 groups: DC control group,Rapa treatment group,3-MA treatment group,PRRSV group,Rapa+PRRSV group Groups and 3-MA+PRRSV groups.First induction of dendritic cells and the determination of PRRSV toxicity,followed by the detection of the effect of PRRSV on the antigen presenting function and autophagy of dendritic cells.Finally,autophagic activators and inhibitors were used to regulate the autophagic target egg m TOR,and then the autophagic target protein m TOR was detected in the PRRSV infection of dendritic cell antigen presentation function.Control.Fluorescence quantitative PCR,inverted fluorescence microscopy,laser confocal microscopy,and Western Blot were used to detect the changes of autophagy and the antigen presentation of dendritic cells.The results showed that PRRSV infection could induce autophagy in dendritic cells and increase the transcriptional level of autophagy related gene Beclin-1(P < 0.01).The transcriptional level of Atg7 increased significantly(P < 0.05),and the transcriptional level of the antigen presenting gene Lamp-1 was significantly reduced.However,there was no significant difference in the transcription level of antigen presenting gene CD40 and TAP-1(P > 0.05).Western-Blot results showed that PRRSV infection promoted the degradation of p62 in dendritic cells and induced the transformation of LC3-I into LC3-II,which not only promoted the formation of autophagic bodies,but also promoted the degradation of autophagic bodies.100 nmo L/L autophagy activator Rapa action 8h and 50 nmo L/L autophagy inhibitor 3-MA action 4H can regulate the activity of autophagic target protein m TOR,and then enhance and reduce the level of autophagy caused by PRRSV,respectively,but it is necessary to further study whether the infection and proliferation of PRRSV can be completely suppressed by this method.