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Objective To study the presence of CD4+CD25+regulatory T cells and hepatitis B virus(HBV)specific cytotoxie T lymphocyte(CTL)in peripheral blood and liver tissues of patients with chronic hepititis B(CHB)and its clincial significance.Methods One hundred and fifty-seven HBV-infected patients,including 20 cases of acute hepatitis B,115 cases of chronic hepatitis B,and 22cases of HBV-related liver cirrhosis,and 20 healthy controls were enrolled in this study.Peripheral blood was collected and liver tissues were obtained from some of the enrolled subjects.The CD4+CD25+regulatory T cells and HBV specific CTL were analyzed using flow cytometry and cytokine flow cytometry(CFC).The comparison between groups was done by t test.Results The percentages of CD4+CD25+ regulatory T cells in the peripheral blood of patients with acute hepatitis B and CHB of mild,moderate and severe degree were(2.87±0.94)%,(3.53±1.56)%,(4.59±2.98)%and(3.65±1.73)%,respectively,which were higher than that of controls(2.36±0.60)%(t=2.04,5.97,3.30 and 3.17,respectively,P<0.01).The percentages of HBV specific CTL in the peripheral blood of patients with mild,moderate and severe degree of CHB and HBV-related liver cirrhosis were (0.189±0.152)%,(0.103±0.110)%,(0.118±0.120)%and(0.098±0.101)%,respectively,which were significantly lower than that of acute hepatitis patients [(0.815±0.360)%](t=10.09,11.87,9.17 and 8.96,respectively,P<0.01).CD4+CD25+ regulatory T cells and HBV specific CTL in liver tissues were both higher than those in the peripheral blood.Conclusion CD4+CD25+regulatory T cells may play an important role in anti-HBV immune response through inhibiting CD8+T eell function.
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Objective To establish a time-resolved immunofluorometric assay (TRFIA) to detect seurm CⅣ(collagenⅣ). Methods The antibodies to CⅣwere coated on mircoplate and the europium-labeled monoclonal antibody of CⅣ. The luminescent enhancement system was used as enhancement solution which contained mainly 2-naphthoy trifluoroacetone. we established A sandwich time-resolved fluoroimmunoassay (TRFIA) was established to measure the seurm CⅣin 127 patients with hepatitis and 30 normal controls. Results The sensitivity of assay was 12. 8?g/L. The coefficient of variation for inner-batch and inter-batch were 4. 54% and 8. 06%,respectively. The recovery was 98. 6%. The serum level of CⅣwas 46. 06?22. 21?g/L in normal control,47. 25?22. 58?g/L in acute hepatitis, 129.01?53.68?g/L in mild chronic hepatitis,277. 90?92.36?g/L in moderate chronic hepatitis,413.90?162.24?g/L in serious chronic hepatitis,568. 60?210.40?g/L in liver cirrhosis. As compared to normal control,higher concentrations of CIV (P