Interaction Between Vesicular Stomatitis Virus And Monocyte And MoDC

Vesicular stomatitis virus belong to the Rhabdoviridae family blisters virus genus,and causes substantial economic losses to the pig industry worldwide.Effective control of the Vesicular stomatitis remains a challenge due to the complicated host-pathogen interactions which lead to delayed immunity,and no suitible vaccine has been available to provent it.Complete virus particles comprise five main structural protein,The matrix protein M has been proved as a major pathogenic determinant of VSV in rodents,but its role in pathogenesis of natural host was still unknown.In order to explore the relationship between the M protein and immune cells,Porcine monocytes were isolated from peripheral blood using MACS technology,than monocytes were treated with GM-CS and IL-4 to obtain the Monocyte-derived dendritic cell.Three recombinant VSV was used to infect the pig monocyte and monocyte derived dendritic cell,include the wild-type VSV-GFP,the VSV with M51 deletion(VSV_ΔM51-GFP)and the VSV with triple mutations with M51 deletion,V221 F,S226R(VSV-MT-GFP).plaque experiment was set up to make growth curve,confocal laser scanning microscope and Flow cytometry are used to assess the cell infection status.The result show that three viruses can be infected monocyte,but the replication of recombinan VSV in Monocyte was generally low,the monocyte infected rate is only8.5% at MOI of 1,while MoDC can be infected by VSV effectively,the concentration of VSV in MoDC supernatant can reach maximum after cells infected 12 h,and then decreased slowly.and they can effectively activate the expression of TNF-α,IL-8,IFN-αin MoDCs.Compared to wild type VSV,the pathogenicity of VSV_ΔM51 and VSV-MTare decreased sequentially.Based on the in vivo and in vitro studies,it was proved that M protein is a virulent factor for VSV in pig and can be an ideal target to develop effective vaccine vectors for swine.Furthermore,in order to establish the method of detection swine IFN-β.BALB/cmouse were immunized by IFN-β by prokaryotic expression.After cell fusion and limited dilution,we obtain three cell lines that could against the IFN-β,the ascites can be verified through the Western Bolting that three mABs reacted specifically with IFN-β,which lay the foundation for assay IFN-β subsequently.