The Expression Of Th17and Treg Cell In Peripheral Blood From Patient With Chronic Severe Hepatitis B

ObjectiveTo explore the impact of Th17cells, Treg cells and their cytokine on the chronicserious hepatitis and chronic severe hepatitis patients.MethodsTwenty six patients with the chronic serious hepatitis B and20patients with chronicsevere hepatitis B from our hospital from January2010to June2012were chosen asresearch subjects; the peripheral blood was collected and the Th17/Treg, Th17/CD4+T andTreg/CD4+T cells from two group were detected using Flow cytometry; the expression ofRORγt and FOXP3mRNA were demonstrated using RT-PCR,and IL-17and TGF-β inculture supernatant of PBMC were tested using ELASE assay; also the impact of Th17andTreg cells on the pathogenesis of chronic serious hepatitis and chronic severe hepatitis wasanalyzed.Results1. Th17/Treg, Th17/CD4+T and Treg/CD4+T in patients with chronic serious hepatitiswere5.18,4.29and5.60, while those in chronic severe hepatitis B patients were2.01,1.08and6.92,there was a significantly statistic difference between the two groups(P <0.05).2. The concentration of RORγt and FOXP3were0.528,0.516μg/μl in the chronicserious hepatitis group and0.504,0.522μg/μl in the chronic severe hepatitis respectively,showing no siginificant difference(P>0.05); The purity of RORγt and FOXP3were1.842,1.881and1.828,1.834respectively, falling in the range of1.8to2.0, demonstrating thetotal RNA concentration and purity of each sample Meeting experimental requirements;the electrophoresis strip presented three clear bands under UV transilluminator, indicatingthe RNA was integrity. The expression of RORγt and FOXP3mRNA in PBMC were1.08and0.70in chronic serious hepatitis B patients0.65and1.18in chronic severe hepatitis Bpatients respectively with significantly statistic difference (P<0.05). 3. The content of IL-17and TGF-β in supernatant of cultured PBMC in patients withchronic serious hepatitis B were41.60pg/ml and256.19pg/ml, and those in chronic severehepatitis B were32.48pg/ml and299.37pg/ml, with significantly statistic differencebetween two groups (P<0.01).ConclusionTh17cells and Treg cells in the peripheral blood of patients with chronic severehepatitis B are activated functionally：Th17/Treg’ imbalance, Th17/CD4+T increasing,Treg/CD4+T decreasing, the RORγt and FOXP3mRNA expression up regulating and theIL-17and TGF-β increasing. All above have an important impact on the liver injury ofpatients with chronic severe hepatitis B.