Study On The Association Between HLA Class Ⅱ, Ⅲ Genes And Viral Hepatitis B

To investigate the association between HLA class II, III genes and viral hepatitis B, We performed the following investigations:1. HLA-DR and HLA-DQ were determined in specimens of 36 cases of chronic hepatitis B and 20 cases of normal liver tissues by immunohistochemistry and Western-blot, and HLA-DR/HBcAg and HLA-DQ/HBcAg were determined by immunohistochemical double-labelling staining to analyse the expression and significance of HLA-DR and HLA-DQ in chronic hepatitis B.2. HSP27, HSP70 and HSP90 a were determined in specimens of 36 cases of chronic hepatitis B and 20 cases of normal liver tissues by immunohistochemistry. HSP70/HBcAg and HSP90 a /HBcAg were determined by immunohistochemical double-labelling staining and HSP70 mRNA was determined by in situ hybridization technique to analyse the expression and significance of HSP27, HSP70 and HSP90 a in chronic hepatitis B.3. HLA-DRB1, HLA -DQA1 and HLA-DQB1 alleles in 54 patients with chronic hepatitis B, 30 patients with acute hepatitis B, 32 patients with chronic severe hepatitis B and 106 normal control subjects were determined by the polymerase chain reaction/sequence specific primer (PCR/SSP) to analyse the association between the polymorphism of HLA-DRB1, HLA-DQA1, HLA-DQB1 alleles and viral hepatitis B.4. Perpheral blood lymphocyte response to recombinant hepatitis B core antigen (rHBcAg) was determined by MTT in 54 patients with chronic hepatitis B and 30 patients with acute hepatitis B to analyse the associationinbetween the lymphocyte proliferation index and the polymorphism of HLA-DRB1, HLA-DQA1 and HLA-DQB1 alleles.5. Immortalized lymphoblastoid cell lines (LCL) were created by EBV infecting peripheral blood lymphocytes in 54 patients with chronic hepatitis B, 30 patients with acute hepatitis B and 32 patients with chronic severe hepatitis B in vitro, and TNF production in LCL was determined by crystal violet staining to analyse the association between the TNF producton in LCL and clinical phenotype of viral hepatitis B and the polymorphism of HLA-DRB1, HLA-DQA1 and HLA-DQB1 alleles .The main results and conclusions were as follows:1. HLA-DR and HLA-DQ were not detected in hepatocytes of normal liver tissues. The positive rates of HLA-DR and HLA-DQ were 22.22% and 19.44% in chronic hepatitis B, respectively. The positive rates of HLA-DR and HLA-DQ in moderate and severe hepatitis ( 37.5%, 37.5% ) were higher than in mild hepatitis ( 10%, 5% ), with significant correlation between them (x 12=3.89, x 22=5.99, PO.05). These findings suggest that overexpression of HLA-DR and HLA-DQ are closely associated with HBV activity and hepatocyte necrosis, and play an important role in pathogenesis of chronic hepatitis B. HLA-DR (HLA-DQ) and HBcAg simultaneously positive hepatocytes obviously denaturalized and necrosed. These findings suggest that both HLA-DR (HLA-DQ) and HBcAg may play a cooperative role in immune-induced damage of chronic hepatitis B.2. The positive rates of HSP27 were 25% and 20% in chronic hepatitis B and normal liver tissues, respectively. There was no significant correlation between them (x 2=0.18, P>0.05). The positive rates of HSP70 and HSP90 a in chronic hepatitis (44.44%, 38.89%) were higher than in normal liver tissues (15%, 15%), with significant correlation between them ( x ,2=4.97, x 22=4.97, P<0.05). The positive rates of HSP70 and HSP90 a in moderate and severehepatitis (68.75%, 62.5%) were higher than in mild hepatitis (25%, 20%), with significant correlation between them ( x 12=6.89, x 22=6.76, P<0.05). HSP70 and HSP90 a mainly expressed in HBcAg negative hepatocytes. These findings suggest that increation of HSP70 and HSP90 a expression may be induced by HBV infection in hepatocytes, but HSP27 not. HSP70 and HSP90 a may be a marker of hepatocyte damage in chronic hepatitis B, and may interfere with production or assemblation of HBV protein.3. Fourteen HLA-DRB1 alleles, ten HLA-DQA1 alleles and thirteen HLA-DQB1 alleles...