| Object①Analyze the relationship between the concentration of FK506 and the level of T-lymPhoey subsets in the patients who accepted liver transplantation as to hepatitis B virus-related terminal diseases.②Study the correlation between peripheral blood mononuclear cell HBV DNA and the level of T-lymphocyte subsets. Investigate the relationship among the three factor to offer the theoretical viewpoint of how to individually use FK506 regime to prophylaxis HBV recurrence after liver transplantation.Method We used fluorescein-labelled monoclonal antibodies and flow- cytometer to determine the T-cell subsets. There were three groups: health volunteers (health-control group, 20 people),patients with chronicity type B hepatitis(CHB) (CHB -control group, 20 people),allo-liver recipients receiving FK506-treatment(FK506-treatment group, 23 people). We collected samples from patients in FK506-treatment group at the ends of 4th week, 8th week, 12th week, 16th week after transplantation.And accordingly called them FK506-treatment 4 week group, FK506-treatment 8 week group,FK506-treatment 12 week group, FK506-treatment 16 week group,and divided into two subgroups in every group calling FK506-high concentration group and FK506-low concentration group respectively.We used SPSS 13.0 to make statistical description and analysis. As for normal distribution ,We used t-test,one-way ANOVA test, Tamhane's T2 test to give comparison between groups. As for nonnormal distribution, we use median and inter-quartile range to describe the data and used Kruskal Walks and Nemenyi nonparametric rank test to do comparison among groups. When p<0.05, the difference was significant.We use multiple linear regression to analyze the correlation of HBV DNA in PBMC and T-lymphocyte subsets after liver transplantation.Result①In FK506-treatment group ,the percentages of CD3+CD4+, CD3+CD8+ and CD8+CD28+ T cells and CD8+CD28+/ CD8+CD28- ratio were much lower than those in health-control group and CHB -control group; inversely ,the percentages of CD8+CD152+ was higher (p<0.05).The percentages of CD8+CD28- in FK506-treatment group was higher than tthat in health-control group(p<0.05) but there were no significant difference between FK506-treatment group and CHB -control group (p>0.05). The FK506 concentration after liver transplantation gradually descended with the time passing (There were significant difference between 4th week and 12th week, 4th week and 16th week, 8th week and 16th week, 12th week and 16th week).The percentages of CD8+CD28+ and CD8+CD28+/ CD8+CD28- ratio gradually increased with the FK506 concentration descent.On the contrary,the percentages of CD8+CD28-,CD8+CD152+ declined by degrees. The percentages of CD8+CD28+ and CD8+CD28+/ CD8+CD28- ratio in treatment 4 week group were significantly lower than those in treatment 12 week group and 12 week group (p<0.05). The very reverse, the percentages of CD8+CD28-, CD8+CD152+ were higher (p<0.05).The percentage of CD8+CD152+ in treatment 8 week group was significantly higher than that in treatment 8 week group and 16 week group.In the treatment 4,8,12week groups, the percentages of CD3+CD4+,CD8+CD28+ and CD8+CD28+/ CD8+CD28- ratio of FK506-high concentration group(≥9ng/ml ) were significantly lower than those of FK506-low concentration group﹙ < 9ng/ml ), and then the percentage of CD8+CD152+ was higher (p<0.05). In the treatment 4,8week groups, the percentage of CD8+CD28-of FK506-high concentration group was significantly higher than that of FK506-low concentration group (p<0.05).②In the treatment 4,8,12week groups,HBV DNA in PBMC of FK506-high concentration group was significantly higher than that of FK506-low concentration group (p<0.05).③With multiple linear regression to analyze the correlation of HBV DNA in PBMC and T-lymphocyte subsets after liver transplantation, we found that HBV DNA intimately correlate with CD8+CD28+, CD8+CD28-, CD8+CD152+,R2= 0.795.According to the result of regression equation,we suggested that HBV DNA had significant postive correlation with CD8+CD28-, CD8+CD152+ and had significant inverse correlation with CD8+CD28+. Conclusion①The effect which the variance of FK506 concentration functioned on CD8+CD28+, CD8+CD28-, and CD8+CD28+/ CD8+CD28- ratio was more sensitive than that on CD3+CD4+,CD3+CD8+.②FK506 concentration mought affect HBV DNA in PBMC .We presumed that the mechanism of HBV recurrence correlated with FK506 concentration after liver transplantation.③FK506 could inhibit the function of T-lymphocytes immune system to eliminate HBV in PBMC by directly suppressing the level of CD3+CD4+,CD8+CD28+ T-lymphocyte subsets and up-regulating the expression of CD8+CD28-, CD8+CD152+ T-lymphocyte subsets to interfere the activation of costimulatory signal molecule.
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