The Expression And Their Clinical Significance Of Toll-Like Receptors In Term Human Placenta Tissues Of HBsAg Carriers

Objective:The aim of this paper is to explore the expression and clinical significance of Hepatitis B surface antigen (HBsAg),Toll-Like Receptor 3 (TLR3),Toll-Like Receptor 4(TLR4),Toll-Like Receptor 7(TLR7) in term human placenta tissues of HBsAg carriers and HBsAg sero-negative pregnant women.And to evaluate the relationship between TLR3,TLR4,TLR7 in human placenta tissues of HBsAg sero-negative pregnant women and HBsAg carriers pregnant women before injection of hepatitis B immunoglobulin(HBIG) and hepatitis B virus serologic markers(HBV-Ms) in peripheral blood from fetuses of HBsAg carrier mothers.Methods:From June 2010 to December 2010,53 cases of HBsAg carriers pregnant women and 10 cases of HBsAg sero-negative pregnant women were collected in the First Hospital Affiliated to Jinan University.The main virus and microorgnization infection of those pregnant women were eliminated. Blood samples (2.5mL) were taken from the peripheral vein from newborns within 12h after birth, before Passive and/or active immunization,and subjected to HBV-Markers detection by ELISA,HBV-DNA qualitative analysis by fluorescence quantitative PCR. The sera were divided and stored at -20℃.At the same time,the placental tissues were also collected. Following studies were preformed:First,the expression of HBsAg,TLR3,TLR4 and TLR7 in term human placenta tissues from HBsAg sero-negative and HBsAg carriers pregnant women were detected by immunohistochemistry;Second,Neonatal serum tested the expression of HBsAg, anti-HBs,HBeAg,anti-HBe,anti-HBc and HB-PreSl were detected by ELISA;Third, the natal sera tested HBV DNA was detected by real-time PCR.Result:1. Pregnant women age,childbirth gestational age,delivery mode and the neonatal weight, gender of HBsAg sero-negative and HBsAg carriers pregnant women were tested,there is no statistical differences in two groups (P>0.05).In HBsAg carriers pregnant women,the expression of HBeAg positive in pregnant women account for 37.7%(20/53);HB-PreS1 positive accounted for 30.2%(16/53);HBV-DNA positive (DNA<103copies/mL for negative group;>103copies/mL positive group) HBV-DNA tested positive for 30.2%(16/53),HBIG intervention in third trimester of application for 13.2%(7/53).The newborns from HBsAg carriers pregnant women mainly expressed HBsAb,HBeAb,HBcAb.The detection rate of HBV-Ms of was 52.8%(28/53), 45.3%(24/53),77.4%(41/53),respectively.Six neoborns HBeAg positive of HBV-Ms accounted for 11.3%(6/53).The loading of HBV-DNA in all neoborns were less than 500copies/mL2. All of placenta tissues from HBsAg sero-negative and HBsAg carriers pregnant women expressed TLR3,TLR4,TLR7,besides one case of HBsAg carriers pregnant women lacking of TLR7.2 cases of HBsAg sero-negative pregnant women and 4 cases of HBsAg carriers pregnant women is deficient of the expression of TLR3 in placenta.The positive signals of TLR3,TLR4, TLR7could locate in the cytoplasm of decidual cell, trophoblastic cells,villous meseachymal cells and villous capillary endothelial cells. The positive signals of HBsAg was mostly located in the villous meseachymal cells and villous capillary endothelial cells. Only 3 cases of placenta showed HBsAg positively in 53 cases. The expression of TLR3,TLR7 in HBsAg-negative placental tissues were significantly higher than the normal pregnant placental tissues(P<0.017). There is no difference in the expression of TLR4(P>0.05).3. The positive signals of TLR3,TLR4,TLR7 in the HBsAg carriers pregnant women placental tissues were stronger than in the HBsAg-negative pregnant women placentas.(t=5.938,P=0.000; t=5.647,P=0.000); There is no difference in the expression of TLR4(t=0.496,P=0.522). The expression and distribution of TLR7 stronger and more widely than TLR3 in the HBsAg carriers pregnant women placental tissues(P<0.05).4. Compared with HBsAg-negative pregnant women,the expression of TLR3,TLR7 was significantly different in human placenta of HBsAg carriers pregnant women which the positive expression of HBeAg,HB-PreSl antigen,and the loading of HBV-DNA more than 103copies/mL(P<0.05). There is no different in the expression of TLR4.5. Compared with HBsAg-negative pregnant women,the expression of TLR3,TLR7 was significantly different in human placenta of HBsAg carriers pregnant women which the administration of HBIG in the third trimester pregnancy (P<0.017). There is no different in the expression of TLR4(P>0.05).6. The expression of TLR3,TLR7 in HBsAg carriers pregnant women, which the newborns positively expressed HBsAb, HBeAg, HBeAb, HBcAb, was different significantly than HBsAg negative group(P<0.05);and there was not statistically different in the expression of TLR4(P>0.05). The expression of TLR3 in HBsAb-positive newborns born of HBsAg carriers pregnant women was significantly higher than HBsAb-negative newborns of HBsAg carriers pregnant women(P<0.05). Conclusion:1. The expression of TLR3,TLR7 in HBsAg carriers pregnant women is significantly higher than the normal non-HBsAg carriers pregnant women,This indicate that the expression of TLR3,TLR7 in placenta perhaps can play an important role in immune regulation,which prevent the vertical transmission of HBV;2. There is no difference between HBsAg carriers pregnant women and normal pregnant women in the expression of TLR4;3. Higher expression of TLR3 in HBsAg carriers pregnant women is closely linked with the expression of HBsAb in newborns which indicated that TLR3 might involve in the neonatal immune regulation of preventing HBV infection.