| ObjectivesBy detecting the peripheral blood T lymphocyte surface costimulatory molecules and T lymphocyte subsets in HBV-infected population,the relationship between HBV viral load and HBV infection and its role in the development of HBV infection were explored.MethodsAccording to the exclusion criteria of this study,120 newly diagnosed HBV patients admitted to our hospitals(The First Affiliated Hospital,School of Medicine,Zhejiang University;Zhejiang Province Youth Hospital)infection department from October 2016 to January 2018 were included in the study.Among them,60 patients with chronic viral hepatitis B(CHB),30 patients with hepatitis B cirrhosis(LC)and 30 patients with acute-on-chronic liver failure(ACLF),CHB group was divided into HBeAg positive group(30 cases)and HBeAg-negative group(30 cases).In the same period,30 healthy volunteers were used as control group to detect the general biochemical indexes,and Elisa was used to detect the serum markers of hepatitis B,and to detect the serum markers of hepatitis B by enzyme-linked immunosorbent assay(Elisa).Flow cytometry was used to detect CD4+and CD8+T cell surface programmed death factor(PD-1),cytotoxic T lymphocyte associated antigen-4(CTLA-4)T cell immunoglobulin mucin molecule-3(Tim3)T cell(CD3+),helper T cell(CD4+)and helper T cell(CD4+).Inhibitory T cell(CD8+)percentage was detected by polymerase chain reaction(PCR).SPSS17.0 statistical processing software was used to analyze the original data.For the measurement data,if it was found to be normal distribution by normal test,the results were expressed in the form of mean±standard deviation(x±s).When the measurement data were compared,the analysis of variance of the completely randomized design data was used,and the HSD method was used to compare the data between groups.If the normal distribution was not satisfied by normal test,the results were expressed in the form of(median,lower quartile to upper quartile)(M,P25-P75),and the two sets of econometric data were compared by rank sum test;the classification variables were expressed in the number of examples(percentage)(N;%)means that if the total sample size was greater than 40 and the minimum theoretical frequency was greater than 5,the chi-square test(?~2 test)was used.If the total sample size was greater than 40 and the minimum theoretical frequency was between 1 and 5,the correction formula of the chi-square test was used.If the total sample size was less than 40,or the minimum theoretical frequency was less than 1,Fisher exact probability method was used.The correlation between the two factors was analyzed by Pearson linear correlation analysis.If the factors did not conform to the normal distribution,the data were transformed to make them conform to the normal distribution,and then the Pearson linear correlation analysis was carried out.There was statistical significance when the level of significant test was 0.05(P<0.05).Results1.The expression of CD4+ CTLA-4 and CD8+CTLA-4 in HBV infected patients waslower than that in control group(P<0.05).The expression of CD4+CTLA-4,CD8+PD1,CD4+Tim3 and CD8+Tim3 in CHB group was significantly higher than thatin LC group and ACLF group(F=56.359,28.364,50.323,56.714,82.774,respectively,P<0.05.Pairwise comparison HSD-q test,P<0.05),the expression of CD4+CTLA-4,CD8+CTLA-4,CD4+Tim3 inACLF group was the lowest,the expression of CD4+PD1 was the highest.The expression of CD4+CTLA-4,CD8+CTLA-4 in HBeAg positive group was lower than that in HBeAg negative group and CD4+PD 1,CD8+PD 1,CD4+Tim3 and CD8+Tim3 expression was higher than that in HBeAg negative group(t=18.536,15.759,20.009,11.228,31.529,23.659,P<0.05,respectively).2.The expression of CD3+CD4+and CD4+/CD8+in HBV infected patients was lower than that in the control group,and the expression of CD 8+was higher than that in the control group.The expression of CD3+CD4+and CD4+/CD8+in CHB group was higher than that in LC group and ACLF group(P<0 05),the expression of CD8+in CHB group was lower than that in LC group and ACLF group(F=22.591,13.665,44.664,32.113,respectively,P<0.05),the expression of CD3+CD4+and CD4+ CD8+in ACLF group was the lowest,and the expression of CD8+in ACLF group was the highest.The expression of CD3+CD4+and CD4+/CD8+in HBeAg positive group was lower than that in HBeAg negative group,and the expression of CD 8+in HBeAg positive group was higher than that in HBeAg negative group.The difference was statistically significant(t=25.227,20.674,19.842,24.516,respectively,P<0.05).3.The HBV-DNA level in CHB group(4.58±0.09)(lg copy/mL)was significantly higher than that in LC group(3.64 ±0.41)(lg copy/mL)and ACLF group(3.48±0.35)(lg copy/r/mL),(F=5.147,P<0.05).The level of HBV-DNA was the lowest in the group of ACLF.The HB V-DNA level of HBeAg positive group(4.99±0.09)(lg copy/mL)was significantly higher than that of HBeAg negative group(4.12±0.08)(lg copy/mL)(F=39.573,P<0.05).4.There were significant positive correlations between HB V-DNA and CD8+PD1,CD8+Tim3 in patients with HBV(r=0.62,0.77,P<0.05),but no correlation with CD4+CTLA-4,CD8+CTLA-4,CD4+Tim3,CD4+PD1,CD3+CD4+and CD8+(r=0.12,0.07,0.09,0.16,0.19,0.11,0.13,P>0.05).ConclusionsThe proportion of costimulatory molecules and T lymphocyte subsets on the surface of peripheral blood T lymphocytes were associated with the development of the disease.The expression of CTLA-4 PD-1 Tim-3 and the proportion of T lymphocyte subsets could be used as an index to evaluate the immune function,and they are closely related to the progress of the disease. |
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