Research On The Relationship Of TLR2/4 On Peripheral Blood Mononuclear Cell In ASC And HBV Proliferation

Objective:To know more about the immune functional state of the ASC after the invading of HBV through the investigation on the relationship of HBV replication and TLR2/4 on Peripheral Blood Mononuclear Cell.Methods:Isolating and collecting PBMCs and sera from ASC.Group them by the status of HBeAg:positive or negative,then select healthy volunteers as controls.Immunocytochemistry was performed to examine the protein expression of TLR2 and TLR4 on the surface of PBMC.Then we detect the mRNA expression of TLR2 by RT-PCR.At the same time we detect the serum TNF-αlevel by ELISA.Results:A,The HBV DNA levels correlate significantly with the status of HBeAg,HBeAg-positive goes with high level of HBV copies(r=0.760, P=0.000).B,After Immunocytochemistry staining,we can see chocolate brown stain on the surface of PBMC membrane in the three group.The positive rate and protein expression of TLR2 on the surface of PBMC in HBeAg positive group are all lower than that in HBeAg negative group and control group show significant difference(P=0.004,P=0.003;P=0.000,P=0.000).The RT-PCR result of mRNA expression level of TLR2 in PBMC consist with the Immunocytochemistry result of protein expression level.The mRNA expression of TLR2 on the surface of PBMC in HBeAg positive group is lower than that in HBeAg negative group and control group show significant difference (P=0.000,P=0.000).But the protein expression of TLR4 on the surface of PBMC in the three groups have no significant difference(P＞0.05).C,The serum TNF-αlevel in HBeAg negative group is higher than that in HBeAg positive group and also the HBeAg positive group is higher than that in control group,they all have significant difference(P=0.000,P=0.000)Conclusion:A,The expression and activation of TLR2 on PBMC in ASC correlate significantly with the HBV proliferation:In HBeAg-positive ASC the replicaiton of HBV DNA is ongoing and with a higher level of HBV DNA copies,they associate with the downregulation of TLR2 on PBMC.On the contrary,the replication of HBV DNA is inactive and with a lower level of HBV DNA copies in HBeAg-negative ASC,they associate with a normal expression and activation of TLR2 on PBMC.B,There is no significant association of HBV replication and proliferation with the expression of TLR4 on PBMC in ASC.