The Functional Study On Immune Cells That Are Activated By CpG ODN In Chronic HBV Infections

Objective:To study the effects of CpG oligodeoxyribonucleotide (ODN) as adjuvant on the immune responses in PBMC,CD4~+ T cell,CD8~+T cell and DC with chronic hepatitis B virus. so as to further explore the immunopathogenesis mechanisms of chronic hepatitis B virus. These information provides a experimental basis of new immunotherapy for HBV.Methods:With the method of density gradient centrifugation, the randomly selected infections were divided into two groups(immune tolerance and immune clearance ) and the control.Isolating PBMC With the method of density gradient centrifugation,and partly purify CD4~+T lymphocyte,CD8~+T lymphocyte and dendritic cell through immunomagneic bead teachenique(IBT), detecting cell surface specific markers of all the lymphocyte were analyzed by FCM .The specific Th1 cell reaction and activity of CTL in immune tolerant phase and in the immune clearance phase that had stimulated by CpG ODN,HBsAg and the Mixture[CpG ODN~+HBsAg] were analyzed by enzyme linked immune spot(ELISOT) and flow cytometry (FCM),to Estimate the activated ability of infections'immune cell by CpG ODN.Results:In different immune state,the statistical analysis of the IFN-γspots of PBMC,CD4~+Tcell and the CD 107'expression of CD8~+Tcell were differently incubated by HBsAg and M[HBsAg~+CpG ODN] have no significant difference(P>0.05);But the result is adverse to the CD 107'expression of CD8~+Tcell in immune clearance group (p=0.008)and the control (p=0.007),and the rate of CD107a had higher than CD107b.In addition, the IFN-γspots of PBMC and pure CD4~+T cell are respectively stimulated by M have significant difference(P=0.000),and the IFN-γspots of PBMC outnumber to CD4~+T cell.Between different immune state,the statistical analysis of the IFN-r spots of PBMC,CD4~+Tcell and the CD 107'expression of CD8~+Tcell were differently incubated by M have no significant difference(P>0.05);the purified ratio of CD4~+T and CD8~+T cell from PBMC are more than 90%.The Purified DC separately include : plasmacytoid dendritic cells(30.28%),myeloid dendritic cell-1(66.50%) and myeloid dendritic cell-2(0.04%).Conclusions:CpG ODN can slightly increase the IFN-γspots of PBMC and Purified CD4~+T cell and the CD 107'expression of CD8~+Tcell,but it have no significant difference in HBsAg and M. the stimulating effect of M to PBMC outweigh CD4~+Tcell.However, CpG ODN can remarkably increase the CD 107'expression of CD8~+Tcell are stimulated by HBsAg, It is very practicable to research the function of DC which are depurated by IBT.