Analysis Of The Relationship Between HBsAg And Serological Markers Of Chronic Hepatitis B And Establishment Of HBsAg-deficient Hepatitis B Mouse Model

Objective:Hepatitis B virus(HBV)infection causes hepatitis B,while chronic hepatitis B(CHB)can develop to cirrhosis and liver cancer,which seriously endanger life and health of human being.Previous reports have shown that there is lack of effective antiviral immune in the chronic hepatitis B virus(HBV)infection.One of the important reasons is hepatitis B virus surface antigen(HBsAg),which was usually described as a negative regulator of antiviral immune responses.In clinical treatment;seroconversion of HBsAg is considered as an important indicator of functional cure of HBV infection,but it is hard to achieve.HBsAg always keeps positive in the whole natural history of patients with CHB.HBsAg not only regulates antiviral responses but also participates in life cycles such as viral packaging.Therefore,whether the level of HBsAg in the natural history of chronic hepatitis B is related to other viral proteins and DNA and various indexes of liver function,whether it directly affects the course of natural history of chronic hepatitis B,is an important scientific and clinical issue worthy of our study.This article intends to clarify these key scientific and clinical issues by analyzing the relationship between HBsAg and serological markers in chronic hepatitis B and establishing a hepatitis B mouse model of HBsAg deficiency.Methods:1.A total of 155 chronic hepatitis B patients were collected from different periods of natural history.Among them,93 were HBeAg positive,62 were HBeAg negative,22.58%were female,and 77.42%were male.The correlation analyze between HBsAg and virological indicators:HBeAg or HBeAb,HBcAb and HBV DNA and liver injury indicators:ALT,AST and GGT and other serological markers:TB,ALB,GLB,PT,INR and PLT.2.HBsAg reading frame was inserted into pAAV-HBV1.2 plasmid by point mutation technique based on the existing high-pressure hydrodynamic transfection method of pAAV-HBV1.2 plasmid.The mutant plasmid The HBsAg-deficient hepatitis B mouse model was constructed.And further compared the normal wild-type plasmid established hepatitis B model mouse to observe the mouse blood and liver in the virological indicators,as well as immune cells in the liver infiltration and so on.Results:1.Among HBeAg-positive male patients,HBsAg was positively correlated with HBeAg and HBV-DNA and negatively correlated with TB and PT.HBsAg was positively correlated with HBeAg,HBV-DNA and PLT in HBeAg-positive female patients.2.In HBeAg-negative male patients,HBsAg was significantly and positively correlated with HBV-DNA,AST and GGT;there was no correlation between female patients.3.Transfection of the mutant plasmid mouse,HBeAg can be detected in serum,but can not detect HBsAg,indicating the successful establishment of HBsAg-deficient hepatitis B mouse.4.The absence of HBsAg makes viral infections more readily cleared,and immune-cell infiltrates in the liver of HBsAg-deficient HBV transfected mouse are less than those of wild-type HBV mouse,whereas the hepatocytes are more susceptible to frosted glassy lesions.Conclusion:1.From the study of patients,sex disparity is not only presented on viral indicators,but also on liver injury and liver function indicators in CHB patients.Furthermore,our work provides detail information for scientific research,and may help establish a novel treatment strategy of hepatitis B,which based on the gender of patients.2.In the part of HBsAg-deficiant mouse,our research offer a novel method using for in vivo study the role of HBsAg,which seems to participate lifecycle of virus and help immunologic escape in chronic situation.