Upregulation of Toll-like Receptor 4 on T Cells in PBMCs Is Associated with Disease Aggravation of HBV-related Acute-on-chronic Liver Failure

Summary： Immune-mediated inflammatory injury is an important feature of the disease aggravation of hepatitis B virus-related acute-on-chronic liver failure （ACLF）. Toll-like receptors （TLRs） have been shown previously to play a pivotal role in the activation of innate immunity. The purpose of this study was.to characterize the TLR4 expression in peripheral blood mononuclear cells （PBMCs） of ACLF pa- tients and its possible role in the disease aggravation. Twelve healthy subjects, 15 chronic HBV-infected （CHB） patients and 15 ACLF patients were enrolled in this study. The TLR4 expression in PBMCs and T cells of all subjects was examined by real-time PCR and flow cytometry. The correlation of TLR4 ex- pression on T cells with the markers of disease aggravation was evaluated in ACLF patients. The ability of TLR4 ligands stimulation to induce inflammatory cytokine production in ACLF patients was ana- lyzed by flow cytometry. The results showed that TLR4 mRNA level was upregulated in PBMCs of ACLF patients compared to that in the healthy subjects and the CHB patients. Specifically, the expres- sion of TLR4 on CD4＋ and CD8＋ T cells of PBMCs was significantly increased in ACLF patients. The TLR4 levels on CD4＋ and CD8＋T cells were positively correlated with serum total bilirubin （TBIL）, direct bilirubin （DBIL）, international normalized ratio （INR） levels and white blood cells （WBCs）, and negatively correlated with serum albumin （ALB） levels in the HBV-infected patients, indicating TLR4 pathway may play a role in the disease aggravation of ACLF. In vitro TLR4 ligand stimulation on PBMCs of ACLF patients induced a strong TNF-α production by CD4＋ T cells, which was also posi- tively correlated with the serum markers for liver injury severity. It was concluded that TLR4 expression is upregulated on T cells in PBMCs, which is associated with the aggravation of ACLF.

Study on the Cellular Molecular Mechanism of Intrauterine Transmission of Hepatitis B Virus

Objective: To study intrauterine transmission of HBV and its cellular molecular mechanism and influence on the fetus. Methods.. A total of 46 coses of pregnant women who suffered from HBV were divided into HBeAg ( + ) and HBeAg ( -) groups. HBV-DNA in serum and peripheral blood mononuclear cells (PBMC-) of 46 cases of pregnant women before delivery was detected by polymerase chain reaction (PCR). After placenta being delivery, HBV-DNA in serum and cord blood mononuclear cells (CBMC) was also detected by PCR. Results.. The total of positive rates of HBV-DNA in serum and PBMC of pregnant women with hepatitis B were 69. 57% (32/46) and 41. 30% (19/46). The positive rates of HBV-DNA in serum of cord blood and CBMC were 56. 52%(26/46) and 21. 74% (10/46) respectively. Among them, the positive rates of HBV-DNA in serum and PBMC of pregnant women with HBeAg ( + ) were 100. 00% (25/25) and 60. 00% (15/25) respectively. The positive rates of HBV-DNA in serum of cord blood and CBMC were 88. 00% (22/25) and 32. 00% (8/25) respectively. The positive rates of HBV-DNA in serum and PBMC of pregnant women with HBeAg (-) were 33. 33% (7/21) and 19.05% (4/21) respectively. The positive rates of HBV-DNA in serum of cord blood and CBMC were 19. 05%(4/21) and 9. 52% (2/21) respectively. The positive rates of HBV-DNA in serum of cord blood and CBMC of newborns were higher in the group of pregnant women with HBeAg ( + ) than those in the group of pregnant women with HBeAg ( -) (P<0. 01 and P<0. 05). There was no HBV-DNA in serum, PBMC and CBMC of normal pregnant women and normal neonates. Conclusion: The intrauterine transmission of HBV can be existent and its transmission way not only can be induced by serum but also can be induced by PBMC. The way of intrauterine transmission of HBV induced by PBMC was concealed. The dangerous possibility of intrauterine transmission is higher in the pregnant women with HBeAg (+) than that in the group of pregnant women with HBeAg ( -).