Preliminary Study On HBV Intrauterine Infection Induced By Hofbauer Cells

Objective To explore the molecular mechanism of HBV intrauterine infection via Hofbauer cells of human villus through pathway of hAV.To isolate and purificate highly pure and active Hofbauer cells in human villus.To explore the effect of anti-hAV against the infection of HBV in Hofbauer cells in vitro.Methods HBVM in serum and supernatant were detected by ELISA. HBV-DNA in serum,peripheral blood mononuclear cells(PBMC),cord blood mononuclear cells(CBMC),Hofbauer cells,tissues,supematant and cleaning solution were detected by PCR.The expressions of CD163,HBsAg,hAV in placenta and HBsAg in fetal liver were detected by the method of streptavidin-peroxidase(SP) immunohistochemistry.Hofbauer cells were isolated and purificated by mechanical method,enzyme tmion digestion law,Ficoll separation and adherent culture,and identificated by the method of streptavidin-peroxidase(SP) immunocytochemical.Results The HBV intrauterine infection rates of mid and late trimester of pregnancy were 8.33%(2/24) and 24.71%(21/85).The HBV intrauterine infection rates of placenta with HBV infection group and without HBV infection group were 44.68%(21/47) and 3.23%(2/62),which was significantly differencetχ~2=27.60, (P＜0.05).Human AnnexinⅤcould be detected in placentas with HBV infection group and normal placentas,and which was more in former than that in later.HBsAg and human AnnexinⅤcould be detected in Hofbauer cells.The level of Hofbauer cells in mid and late trimester of pregnant placenta with HBV intrauterine infection were 24.5±2.5 and 39.5±2.7,and those of without HBV intrauterine infection were 13.2±3.2 and 12.3±2.8,which were significantly differencet(P＜0.05).The intrauterine infection rate of neonate with Hofbauer cells HBsAg(+) was 61.29%(19/31),which was more higher than that of HBsAg(-)(χ~2=35.12,P＜0.05).The purity and activity of Hofbauer cells were more than percent of ninty by the union separation.Hofbauer cells which were be infected by HBV were be washed.The first time weakened in turn to the 11th cleaning solution intensity.The 12th cleaning solution HBV-DNA was negative.Expression of HBsAg and HBV-DNA in supematant were positive after raised 36h.Expression of HBV-DNA in Hofbauer cells was negative after raised 24h by HBV positive serum with anti-hAV.Expression of HBsAg and HBV-DNA in supernatant were negative after raised 72h.Conclusion HBV can infect pregnant women and their fetal in the early,mid and late trimester of pregnancy.HBVcan infect placenta and Hofbauer cells by the way of hAV,which plays a key role on infection of HBV intrauterine infection.The high purity and activity of Hofbauer cells could be obtained by the union separation. Hofbauer cells could be infected by HBV positive serum whitout other assistant and some viral particle of HBV could be released,hAV,a receptor on surface of Hofbauer cells,plays a key role on infection of HBV.Anti-hAV may effectively prevent HBV invading into Hofbauer cells.