| BackgroundThe development of hepatitis B virus transgenic mice advance the study of pathogenesis of chronic liver disease.,and many economical model was developed in the world. C57-TgN (HBVadr 2.0) SMMU was established in 1993 by Department of Cell and Biology in our school. Previous study showed that hepatitis B virus gene was integrated into C57-TgN (adr 2.0) SMMU transgenic mice,and express HBsAg, HBcAg, X protein in liver tissue. HBV DNA were replicated in serum of transgenic mice, HBsAg and HBeAg can be measured in serum but anti- HBsAg and anti- HBeAg wer not found. Partical like HBV can be found in liver tissue and serum It is different from previous study that pathological change was found in the liver of C57-TgN (HBVadr 2.0) SMMU transgenic mice.ObjectiveTo make histological and immunopathological observation on the liver of C57-TgN (HBV adr 2.0) SMMU,and try to provid data for the study on application to HBV transgenic mice.Methods1. The HBV transgenic mice of C57-TgN line in SPF (Specific Pathogen Free) grade(168 case) and corresponding control mice C57BL/6 (15case) were chosen for the study. The liver was taken for histopathologic examination under light microscopy after Hematoxylin-Eosin stain. All of the mice were classified to different groups according to histopathologic examination,and the correlation of liver injury to age, sex were studied.2. Argyrophilic fiber stain was performed to observe intrahepatic fibrosis.3. HBsAg, HBeAg in the serum of HBV transgenic mice was performed by ELISA, and the correlation of liver injury to the level of HBsAg, HBeAg in the serum were studied;4. Immunohistochemical location of HBsAg, HBeAg, X protein in liver tissue was studied to show whether HBsAg, HBeAg, X protein were correlation to liver injury;5. Measuring of subsets of T lymphocyte: CD3, CD4 CD8 in the blood of TGM was performed by flow cytometry.6. To study whether HBV specific intrahepatic celluar immune response causes the liver injury, 20 cases with intrahepatic mononuclear cell infiltrates were chosen to study the immunohistochemical location of subsets of T lymphocyte: CD3 CD4 CD8andCD57in liver tissue and human hepatitis B.Resultl.The liver was taken for histopathologic examination under light microscopy after Hematoxylin-Eosin stain. Comparing with control mice .pathological lesion similar to human chronic hepatitis B was found in 37.5% ,while 32.7% of HBV transgenic mice had mild nonspecific abnormalities. The frequency of liver lesion similar to hepatitis B was significantly correlative with age, pathological lesion of liver occur in transgenic mice older than 6 months.2. Argyrophilic fiber stain show that few fibrosis was found in most cases.3. The expression of HBsAg HBcAg and X protein was found in 77.4% 56.5% 34.8% of transgenic mice, There are minor differences in the expression of HBsAg HBcAg X protein between the liver lesion and the normal, even between the injured and the normal tissue in the same liver,which imply that the pathological change in the liver was not caused by the direct effect of HBsAg HBcAg and X protein on hepatocyte.4. HBsAg was expressed in the serum of 55. 2% of TGM, 25.0% expressed HBeAg in the blood, The OD value of HBsAg and HBeAg in the serum of HBV transgenic mice that have liver injury is lower than the normal control.5. Measuring of subsets of T lymphocyte by flow cytometry show that the expression of CD3 CD4 CD8in the blood of TGM was weaker than the normal mice.6. Most of the intrahepatic mononuclear cell infiltratesare are CD3+CD4+T lymphocyte, but CD8+ CD57+cells were not found.In human hepatitis B.all the marker above were expressed strongly.Conclusion1 .Pathological lesion similar to human chronic hepatitis B was found in C57-TgN (adr 2.0) SMMU transgenic mice.2.Histopathological changes similar to human chronic hepatitis B is correlative with age, but not with the expression and location of HBsAg HBcAg and X protein.3.The level of HBsAg HBeAg in the serum of HBV transgenic mice that have liv... |
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