Hla Alleles And Mhc-peptide-specific Ctl In Viral Hepatitis B Mechanism

PART ONERELATIONSHIP BETWEEN THE ALLELES OF HLA-DRB1 AND EVERY KIND OF PROGNOSIS OF CHINESE PATIENTS OF HBV INFECTION[Aims] To investigate the association between the alleles of HLA-DRB1 and the patients including chronic hepatitis B , carriers with hepatitis B virus and subjects who had spontaneously recovered from HBV infection in Shandong area. So we can acquire the knowledge of the genetic background of CHB patients whith every kind of prognosis .[Methods] The antigens and antibodies of HBV were detected by ELISA. The quantity of HBV-DNA was measured by Roche LightCycler fluorescent PCR detector. Liver founctions (including serum aminotransferase, ALB and GLB) was tested by the automatic biochemical analyzer. HLA-DRB1 alleles were detected by the polymerase chain reaction/sequence specific primer (PCR/SSP) technique.[Results] ① According to the level of serum antigens and antibodies of HBV and HBV DNA ,We subdivided the patients into two groups , high HBV replication group and low replication group. Then we compared the frequencies of HLA-DRB1 alleles fouded that the frequency of HLA-DRB1*0701 in high replication group was much higher than that in low replication group (45% vs 12.5%, P<0.05). The frequencies of the rest HLA-DRB1 alleles in the two groups had no significantly difference (P>0.05).② The four alleles, HLA-DRB1*0701, HLA-DRB1 * 1001, HLA-DRB1 * 1201 and HLA-DRB1*1501 ,are significantly higher in the group of chronic hepatitis B than those in the normal control subjects (23.3% vs 4% , 16.7% vs 2%, 60%vs4%, 56.7%vs12%;P<0.05 s PO.05 n P<0.01 n PO.01), Subdividing the group of chronic hepatitis B into two groups including hepatic cirrhosis and primary hepatic carcinoma (PHC) caused by HBV, we found that the frequencies of HLA-DRB 1 * 1001 and HLA-DRB 1 * 1201 in the patients with hepatic cirrhosis are far higher than those in the normal control subjects (30% vs 2% -. 50% vs 4% , P=0.01 , P=0.04). In the group of primary hepatic carcinoma (PHC), the frequency of allele of HLA-DRB 1 * 1501 is higher than that in the normal control subjects (60% vs 12% , P=0.002).(3) Three alleles, HLA-DRBl*0701 > HLA-DRB1*12OK HLA-DRB1* 1501, are significantly higher in the group of asymptomatic HBV carriers (23.3% vs 4% n 60%vs2% > 56.7%vs 12%, PO.05 > PO.01 , P<0.01).? In the group of subjects who had spontaneously recovered from HBV infection, there is no difference about the allele compared with normal control subjects.[Conclusion] The four alleles, HLA-DRB1 *0701 > HLA-DRB1 * 1001 , HLA-DRB 1*1201 and HLA-DRBl*1501, are the susceptive genes in patients with chronic hepatitis B and the allele of HLA-DRB 1*0701 have relationship with high replication of HBV. The another two alleles of HLA-DRBl*1001 and HLA-DRB1*12O1 are the susceptive genes in the patients with hepatic cirrhosis caused by HBV. The allele of HLA-DRB 1 * 1501 is all so the susceptive gene in patients with primary hepatic carcinoma. The three alleles of HLA-DRBl*0701, HLA-DRB1*12O1, HLA-DRBl*1501 are the susceptive genes in the asymptonstic HBV carriers. In this study we did not find protective or resistant gene to HBV infection.PART TWOTHE SIGNIFICANCE OF HLA-A GENOTYPES AND T CELL SUBCLASSIFICATION OF PATIENTS WITH HBV INFECTION AND THE RELATION BETWEEN THEM[Aims] To study the frequency of different genotypes of HLA-A gene of the patients with HBV infection and their relations between HLA-A n T cell subclassification and liver diseases.[Methods] HLA-A genotype was measured in 30 cases of HBV chronic infection and 50 healthy subjects with polymerase chain reaction-sequence specific primer(PCR-SSP).All the patients and healthy subjects were residents of Jinan and belonged to Han Nationality.And they were analyzed with T cell subclassification.The relationship of different genotypes of HLA-A with the clinical types of HBV infection was analyzed.[RESULTS] The frequency of HLA-A*0201 genotype is 52% in the healthy residents of Jinan with. The frequency of HLA-A*0201 in patients with chronic hepatitis B is 23.3%, but in healthy subjects the frequency is 52% . The frequency of HLA-A*3 is 33.3% in patients with chronic hepatitis B compared to 20.8% in healthy subjects to (P<0.05). Patients with HLA-A*0301 + can be found CD4/CD8 decrease from their PBM .The relative risk coefficient was 0.28 for HLA-A*0201 and 2 for HLA-A*0301. The HLA-A* 1101 was usually found in asymptomatic HBV chronic carrier. Patients with HLA-A* 1101+ can be found CD4/CD8 decrease from their PBM .HLA-A*0201 was increased in patients that recoverdfrom acute HBV infection. Patients with HLA-A*0201 + can be found an CD4/CD8 increase from their PBM .[CONCLUSION] HLA-A*0201 > HLA-A*0301 is related to hepatitis B in JiNan,HLA-A* 1101 is related to carrier with HBV .And the immune system and genetic factors participate in the pathogenesis of hepatitis B. Perhaps HLA-A*0201 ^ HLA-A*0301 -.HLA-A* 1101 is some of the powerful region of HLA-A. They participate in the pathogenesis ofhepatitis B by cytokine and CD4/ CD8.PART THREESTUDIES ON THE ROLE OF CIRCULATING HBcAg SOECIFIC CD8+ T CELL AND RELATED CYTOKINE IN THE PATHOGENESIS OF CHRONIC HEPATITIS B PATIENTS[Objective] To study the role of HBcAg specific CD8+T cell and related cytokines in pathogenesis of chronic hepatitis B(CHB) by analyzing the correlations between which and the serum ALT and HBV-DNA levels .[ Methods 1 HBcAg specific CD8+ cells in whole blood samples of chronic hepatitis B patients were stained with soluble HLA-A2 — HBV core 18-27 peptide tetramer complex and counted by flow cytometry. In the mean time, the serum HBVDNA copies and alamine transaminase(ALT) levels were examined and their relationship were analyzed by statistical strategy.[ Results ] ? The medians of percentages of HBcAg specific CD8+T cells of total CD8+ cells were 0.03 %(0.00 % ~ 0.42%) in 34 HLA-A2+CHB patients and had significant differences comparing with those of HLA-A2+AHB(0.01%,0.01%-1.17%), HLA-A2-CHB(0.01%,0.01%-0.05%)and HLA-A2+Controls(0.02%,0.00%-0.05%).? No any specific correlations were found between the percentages of HBcAg specific CD8+ cells and the serum HBVDNA copies and ALT levels through spearman's rank relation analysis and protracting scatterplots. But 34 caces HLA-A2+CHB patients were further divided into several groups according to the serum HBV-DNA level and the frequences of blood Tel 8-27 peptide specific CTL of which were examined respectively. The results showed that only the frequences of Tc 18-27 peptide specific CTL of the HB VrDNA =§ 1 OOOcps/mlgroup(0.055,0.05 ~ 0.06),the 105 ~ 1006cps/ml group (0.09,0.01 ~ 0.22) and 106 ~ 107cps/ml group(0.03,0.01 ~ 0.42) were significantly higher than those of the control group ( X 2=4.845 ^ 6.3^ 4.903, P < 0.05 ) . In the mean time,the the frequences of blood Tel 8-27 peptide specific CTL also were compared between HbeAg+ group and HbeAb+ of the 34 HLA-A2+CHB and there were no significant between them ( P > 0.05 ) ,but the frequence of HbeAb+group were significantly higher than the controls ( X 2=6.188,P < 0.05)?The serum IFN Y and IL-12 level of 34 HLA-A2+CHB patients ( 73.1400 ± 8.85813, 42.0076 ± 38.3818 ) and of 14 HLA-A2+liver sclerosis patients ( 71.2730 ± 7.24752 , 43.1443 + 45.3180 ) had significant differences comparing with those of controls (59.79408 ±7.01781, 14.5542+ 7.1773 ) , while had no significance with those of HLA-A2+AHB patients (66.36060 + 5.91535 , 22.5400 ± 13.1969 ) .@ 34 HLA-A2+CHB patients were further divided into three groups according to theseverity of disease and the serum IFN Y and IL-12 level of which were examined respectively. The results showed that the serum IFN Y and IL-12 level of the moderate group(MG,76.88595 ± 7.54990 , 43.2727 + 42.7835) and severe group(SG,70.48600 ± 4.21136 , 55.0400 ± 37.7310) were significantly higher than those of the light group(LG,61.61667 ± 4.52354 , 26.5083 ± 11.6788) and controls(59.79408 ± 7.01781 , 14.5542 ± 7.1773) ( P < 0.01 iP < 0.05 ) , while there was no significant differences between the level of MG and SG( P > 0.05 ) .(§) Positive correlations were found between the the serum IFN Y and IL-12 level and the serum ALT level of HLA-A2+CHB patients through spearman's rank relation analysis ( rs=0.307 , 0.412, 0.331, P < 0.01 ^P < 0.05 ) .[ Conclusion 3 (D HBcAg specific CTL is a protective element of CHB patients andreflects the level of specific cell immune capacity in certain degree.(2) The level of HBcAg specific CTL has not any direct correlations to hepatocyte injury. But moderate stimulus of virus antigen is the necessary condition of effective anti-virus cellimmunity.(3) The moderate expression of IL-12 can suppress HBV replication, but its over expression may lead to the spread and persistence of HBV contraction.The degeneration of IL -12 secretion capacity and Th2 type immune reaction mainly in body may be one of the mechanics of chronicHBV infection.? The serum level of IFN Y and IL-12 has positive correlation with hepatocyte injury. The large amount of IL-12 induce Thl type immune reaction and lead to hepatocyte injury and the virus replication recuction, which is one of the reasons of active liver injury in chronic HBV infection patients.