The Quantititive Determination And Its Significance Of Specific CTL And Related Cytokine Of HBV Infector

[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 HBVDNA levels .[Methods] HBcAg specific CD8+ cells in whole blood samples of chronic hepatitis B patients were stained with soluble HLA-A2—HBV core 18-27 peptide tetrameric complex 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] ①The 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 , 0. 04% (0. 01-0. 35% )in 14 HLA-A2+AsC, 0. 65% (0. 01-1. 17% )in 5 HLA-A2+AHB patients and 0.02% (0.00-0.05%) in 12 controls. In the mean time, comparing between groups indicates that HBV sCTL level of HLA-A2+ AHB patients was significantly higher than those of HLA-A2+CHB patients (P <0.01) as well as HBV sCTL level of HLA-A2+CHB patients, AsC and AHB patients was significantly higher than those of controls (P<0.05) . ② No any specific correlation was found between the percentages of HBcAg specific CD8+ cells and the serum HBVDNA copies and ALT levels throughspearman' s rank relation analysis and protracting scatterplots. But 34 HLA-A2±CHB patients were further divided into several groups according to the serum HBVDNA level and the frequences of blood Tc18-27 peptide specific CTL of which were examined respectively. The results showed that only the frequences of Tcl8-27 peptide specific CTL of the HBVDNA(?) 1000cps/ml group(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 (P< 0.05) . In the mean time, the frequences of blood Tcl8-27 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 (P<0.05) ③The serum IFNγ and IL-12 level of 34 HLA-A2±CHB patients (73. 14 ± 8.86, 42.01 ± 38.38) and of 14 HLA-A2±chronic asymptomatic HBV carrier(AsC) (75.27 ± 7.25, 43.14 ± 45.32) had significant differences comparing with those of controls (59. 79 ± 7. 02, 14. 55 ± 7. 18) , while had no significance with those of HLA-A2±AHB patients (66.36 ± 5.92, 22.54±13.20) . ④34 HLA-A2±CHB patients were further divided into three groups according to the severity of disease and the serum IFNr 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, 76.89 ± 7.55 , 43.27 ± 42.78) and severe group(SG, 70. 49±4. 21, 55. 04±37. 73) were significantly higher than those of the light group(LG, 61.62 ± 4. 52, 26.51 ± 11.68) and controls (59.79 ± 7. 02, 14. 55 ± 7. 18) (P< 0. 01 或 P<0. 05) , while there was no significant difference between the level of MG and SG (P>0.05) . ⑤Positive 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 或 P<0.05).[Conclusion] ①HBcAg specific CTL is a protective element of CHBpatients and its level to certain degree reflects the high or low of specific cell immune capacity. ②The level of HBcAg specific CTL has not any direct correlations to introhepatic HBVDNA replication and hepatocyte injury. But moderate stimulus of virus antigen is the necessary condition of effective anti-virus cell immunity. ③The conclusions that the serum IL-12 level of HBV infectors is significantly higher than those of controls and CHB higher than those of AHB and that the serum level of IFN y and IL-12 has positive correlation with hepatocyte injury suggest that the large amount of IL-12 induce Th1 type immune reaction and lead to hepatocyte injury and the virus replication reduction, which is one of the reasons of active liver injury in chronic HBV infection patients.