| Obje ctive: To investigate the serum cytokines profile characteristics in chronicHBV infected patients with different clinical phenotypes and the diagnosis value ofCCL20serum level in HBV-related fibrosis, cirrhosis and hepatocellular carcinoma.Methods: According to the concentration of HBsAg, HBV DNA, HBV serummarkers and ALT levels, we divided90cases of chronic HBV infected patients into twoparts: persistently normal ALT Levels and chronic hepatitis B. Persistently normal ALTlevels part included immune tolerance, inactive HBsAg carrier, HBsAg clearance threephenotypes, healthy blood donor as control; According to the HBeAg status chronichepatitis B was divided into HBeAg positive and HBeAg negative two phe notypes. Bycustomized protein chip, expression levels of30serum cytokines including IFN-gamma,IL-1β, IL-2, IL-4, IL-6, IL-7, IL-9, IL-10, IL-12p40, IL-12p70, IL-15, IL-17A, IL-17C,IL-21, IL-22, IL-23p19, IL28A, IL-29, CCL5, CCL16, CCL20, CCL22, CXCL9,CXCL10, CXCL11, CD30, GITR, IL-6R, gp130and TGF-β1were measured; proteininteraction analysis tool STRING was used for analyzing interaction between thesecytokines. On this basis, we selected CCL20in further analys is. Patients including85cases of chronic HBV infection and45cases of HBV related HCC patients were enrolled,ELISA to measure serum levels of CCL20, Spearman’s rank test to calculate coefficientsof correlation between CCL20serum levels and indexes reflecting liver lesion severity(serum liver fibrosis laboratory and clinical parameters, liver stiffness measurement byFibroscan, pathologic fibrosis stage by liver biopsy, tumor size).The receiver operatingcharacteristic(ROC) curve analysis was used to calculate the optimal cutoff value ofCCL20in diagnosing HBV-related cirrhosis and hepatocellular carcinoma. All statisticalanalyses were performed using PASW18.0. Results:1.Between chronic HBV infected patients with persistently normal ALT and healthycontrols, significant differences of expression levels were observed in ga mma-chaincytokines(IL-2, IL-4, IL-7, IL-9, IL-15and IL-21),IL-12family members(IL-12p40, IL-12p70and IL-23p19) and IFN-λ(IL-28Aand IL-29). The lowest level was in healthycontrols. Among three HBV infected groups, the significant characteristic of inactiveHBsAg carrier group differ from other two groups were upregulation of IL-2, IL-4, IL-12p70, IL-15, IL-21, IL-23p19, and IL-29. All cytokines with significant differenceswere involved JAK-STAT signaling pathway.2. Significant positive correlation between ALT and cytokines, such as CXCL9,CXCL10, CXCL11, CCL20, IL-10, IL-4, IL-17C, IL-1β, IL-12p70, IL-17A, wasobserved in both HBeAg positive and negative patients with chronic hepatitis B, in whichthree CXC chemokines CXCL9, CXCL10, CXCL11had higher correlation coefficient.The phenomenon of significant positive correlation between gp130, IFN-γ, IL-23p19,IL-12p40, IL-22, IL-29, IL-6, IL-9, GITR and ALT was only discovered in HBeAgpositive chronic hepatitis B; significant correlation between IL-7, IL-21, TGF-beta1andALT was restricted to HBeAg negative chronic hepatitis B. The ALT>5xULN groupscomparing with inactive HBsAg carrier group, the numbers of different expressioncytokines were11and17,of which11cytokines in both phenotypes of CHB group, theother6cytokines were only in HBeAg negative chronic hepatitis B group with lowerexpression of IL-29, IL-6, IL-9, IL-6R, IFN-γ, TGF-β1.3.In HBeAg positive chronic hepatitis B patients the correlation coeffic ients ofCCL20serum levels and liver fibrosis indexes(HA, LN, PCⅢ, CⅣ) were0.444(P=0.008),0.396(P=0.019),0.424(P=0.011) and0.415(P=0.013) respective ly, while inHBeAg negative chronic hepatitis B patients these results were0.731(P=9.930x10-6),0.634(P=2.932x10-4),0.361(P=0.218) and0.218(P=0.134) respectively, and thecorrelation coefficient related to liver stiffness measurement was0.507(P=7.282x10-7).4. The area under ROCs (AUROC) of CCL20in diagnosis for HBV-related cirrhosisand poor prognosis HCC were0.848(P=7.055×10-5) and0.718(P=0.03) respectively,while the optimal cutoff value were77.4pg/mL (70%sensitivity and96%specific ity) and66.9pg/mL (72.7%sensitivity and64.3%specificity) respective ly. Conclusions:1. The upregulation of JAK-STAT cytokines is an important immune feature forchronic HBV infection. High expression of IL-2, IL-15, and IL-21, IL-12p70, IL-23p19and IL-29are beneficial for HBV immune clearance.2. CXCL9, CXCL10, CXCL11and CCL20can serve as immune indexes evaluatingdegree of inflammation in chronic HBV infected patients. The upregulation defect of IL-29, IL-6, IL-9, IL-6R, IFN-γ and TGF-β1may correlate with immune pathogenesis inHBeAg negative chronic hepatitis B.3. The CCL20plays a dual character of proinflammation and fibrosis developmentin chronic hepatitis B patients. It is suitable for diagnosis and prognosis immune indexfor HBV-related cirrhosis and hepatocellular carcinoma. |
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