Effect of IL-18 on peripheral blood mononuclear cells of chronic hepatitis B and hepatitis B virus DNA released by HepG2.2.15 cell lines

BACKGROUND: Interleukin-18 (IL-18), a pro-inflamma- tory cytokine that induces interferon-γ (IFN-γ) production in T cells and natural killer cells, plays a critical role in the T-lymphocyte helper type 1 ( Th1) response. This study was designed to explore the effect of IL-18 on peripheral blood mononuclear cells ( PBMCs) derived from chronic hepatitis B (CHB) and on hepatitis B virus (HBV) DNA released by HepG2.2.15 cell lines, which were transfected with hepatitis B virus gene in vitro. METHODS: PBMCs isolated from 25 healthy people and 25 patients with CHB were stimulated with HBcAg and IL-18 of various concentrations for 72 hours. The levels of IFN-γ in the supernatants of cultured PBMCs were determined by ELISA. After the stimulation of IL-18 of various concentra- tions, PBMCs derived from one patient were co-cultured for 96 hours with HepG2. 2. 15 cells which had been cul- tured for 24 hours, and then the supernatants were collected by centrifugation and used for HBV DNA quantitative as- say. RESULTS: When PBMCs were stimulated by HBcAg and IL-18 at various concentrations, the levels of IFN-γ in the supernatants of CHB groups were much higher than those in normal control groups, at 0.2 ng/ml: t =11.70, P< 0.01; at 1.0 ng/ml: t =16.19, P<0.01; and at5.0 ng/ml: t =20.12, P <0.01. In the CHB groups, the levels of IFN-γ in the supernatants of PBMCs stimulated by HBcAg alone were lower than both those stimulated by HBcAg and EL-18 at various concentrations and those stimulated by HBcAg and EL-18 (5.0 ng/ml) together with EL-12 (mild: t = 2.20, P<0.05; moderate; t=2.97, P<0.05; severe; t = 0.66, P >0.05). The content of HBV DNA in the superna- tant of co-cultivation of HepG2. 2. 15 cells and PBMCs without stimulated materials was higher than that stimula-ted by HBcAg and EL-18 at various concentrations of HBc- Ag and IL-18 together with IL-12/IFN-α1lb. CONCLUSION: DL-18 can induce IFN-γ secretion and pro- bably play a key role in the modulation of both innate and adaptive immunity. It has implications in improving im- munoregulatory effect and increasing the ability of immune cells to kill cells infected by virus.

High levels of serum interleukin-6 increase mortality of hepatitis B virus-associated acute-on-chronic liver failure

BACKGROUND Patients with hepatitis B virus-associated acute-on-chronic liver failure(HBVACLF)present a complex and poor prognosis.Systemic inflammation plays an important role in its pathogenesis,and interleukin-6(IL-6)as a pro-inflammatory cytokine is related with severe liver impairment and also plays a role in promoting liver regeneration.Whether serum IL-6 influences HBV-ACLF prognosis has not been studied.AIM To determine the impact of serum IL-6 on outcome of patients with HBV-ACLF.METHODS We performed a retrospective study of 412 HBV-ACLF patients.The findings were analyzed with regard to mortality and the serum IL-6 level at baseline,as well as dynamic changes of serum IL-6 within 4 wk.RESULTS The serum IL-6 level was associated with mortality.Within 4 wk,deceased patients had significantly higher levels of IL-6 at baseline than surviving patients[17.9(7.3-57.6)vs 10.4(4.7-22.3),P=0.011].Patients with high IL-6 levels(>11.8 pg/mL)had a higher mortality within 4 wk than those with low IL-6 levels(≤11.8 pg/mL)(24.2%vs 13.2%,P=0.004).The odds ratios calculated using univariate and multivariate logistic regression were 2.10(95%confidence interval[CI]:1.26-3.51,P=0.005)and 2.11(95%CI:1.15-3.90,P=0.017),respectively.The mortality between weeks 5 and 8 in patients with high IL-6 levels at 4 wk was 15.0%,which was significantly higher than the 6.6%mortality rate in patients with low IL-6 levels at 4 wk(hazard ratio=2.39,95%CI:1.05-5.41,P=0.037).The mortality was 5.0%in patients with high IL-6 levels at baseline and low IL-6 levels at 4 wk,7.5%in patients with low IL-6 levels both at baseline and at 4 wk,11.5%in patients with low IL-6 levels at baseline and high IL-6 levels at 4 wk,and 16.7%in patients with high IL-6 levels both at baseline and at 4 wk.The increasing trend of the mortality rate with the dynamic changes of IL-6 was significant(P for trend=0.023).CONCLUSION A high level of serum IL-6 is an independent risk factor for mortality in patients with HBV-ACLF.Furthermore,a sustained high level or dynamic elevated level of serum IL-6 indicates a higher mortality.

Interleukin-10 promoter polymorphisms in patients with hepatitis B virus infection or hepatocellular carcinoma in Chinese Han ethnic population

BACKGROUND: Since single nucleotide polymorphisms (SNPs) can serve as gene markers, polymorphism profiles may help scientists to identify the full collection of genes that contribute to the development of complex diseases such as cancer. The distribution of interleukin-10 (IL-10) promoter polymorphisms in Chinese Han ethnic patients with hepatitis B virus (HBV) infection and hepatocellular carcinoma (HCC) was investigated in this study. METHODS: The polymorphisms of IL-10 promoter region were detected by pulymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and sequencing. Sixty-six health controls, 42 patients with HBV infection, 30 HCC patients, and cell line SMMC-7721 were examined this way. RESULTS: Polyrnorphisms of T/C or T/N on-872 site occurred frequently in Han ethnic population. Pulyrnorphisms were detected in HBV and HCC patients and cell line SMMC-7721. The hotspot among the pulymorphisms was inserting base A between-1058 and-1057. CONCLUSION: Polymorphisms of IL-10 promoter in HBV and HCC patients may be associated with HBV infection and HCC development.

Relationship between interleukin-6 polymorphism and susceptibility to chronic hepatitis B virus infection

AIM:To identify the relationship between tag single nucleotide polymorphisms(tag SNPs) of interleukin-6(IL-6) gene and susceptibility to chronic hepatitis B virus(HBV) infection in a Han Chinese population.METHODS:We performed a case-control study of501 Chinese patients with chronic HBV infection and301 self-limiting HBV-infected individuals as controls.Genomic DNA was isolated from the whole blood of all subjects using phenol/chloroform with MaXtract highdensity tubes. Tag SNPs were identified using genotype data from the panel(Han Chinese in Beijing) of the phase II HapMap Project. Four tag SNPs in IL-6(rs17147230A/T,rs2066992G/T,rs2069837A/G and rs2069852A/G) were genotyped by the Multiplex Snapshot technique. The genotype and allele frequencies were calculated and analyzed.RESULTS:Five haplotypes were involved in the analysis,with frequencies higher than 0.03. One of the haplotypes,TTAA,was significantly different between the two groups. Overall haplotype P values were:ATAA,P = 0.605,OR(95%CI) = 1.056(0.860-1.297); TGAG,P = 0.385,OR(95%CI) = 1.179(0.813-1.709); TGGG,P = 0.549,OR(95%CI) = 1.087(0.827-1.429); TTAA,P = 0.004,OR(95%CI) = 0.655(0.491-0.873); TTAG,P = 0.266,OR(95%CI) = 1.272(0.832-1.944). However,the four SNPs showed no significant genotype/allele associations with susceptibility to chronic HBV infection. Overall allele P values were:rs17147230,P = 0.696,OR(95%CI) = 1.041(0.850-1.276); rs2066992,P = 0.460,OR(95%CI)= 1.090(0.868-1.369); rs2069837,P = 0.898,OR(95%CI) = 0.983(0.759-1.274); rs2069852,P = 0.165,OR(95%CI) = 0.859(0.693-1.064). Overall genotype P values were:rs17147230,P = 0.625; rs2066992,P= 0.500; rs2069837,P = 0.853; and rs2069852,P =0.380.CONCLUSION:The four tag SNPs of IL-6 gene may be associated with susceptibility to chronic HBV infection in the Han Chinese population.

Serum and ascites levels of macrophage migration inhibitory factor, TNF-α and IL-6 in patients with chronic virus hepatitis B and hepatitis cirrhosis

Objective: To study the potential role of macrophage migration inhibitory factor (MIF), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the development of chronic virus hepatitis B (CH) and hepatitis cirrhosis (HC). Methods: The serum concentrations of MIF, TNF-α and IL-6 in 18 patients with chronic virus hepatitis B and in 14 patients with hepatitis cirrhosis without as- citic fluid, and the serum and ascites cytokine con- centrations in 22 HC patients with ascitic fluid were detected by enzyme linked immunity sorbed assay. Results: The cytokine concentrations of the patients were significantly higher than those of the controls. The serum levels of MIF, TNF-α and IL-6 of the 22 patients with ascitic fluid were higer than those of 14 HC patients without ascites. In the 18 patients with CH, the serum cytokine concentrations were the low- est. The serum cytokine concentrations of the 22 HC patients with ascites were significantly higher than those of the 14 HC patients without ascites (P< 0. 01). Their serum cytokine concentrations were sig- nificantly higher than those in the 18 patients with CH (P<0. 01). The concentration of IL-6 in ascites was the highest among all the groups. The serum le- vels of MIF, TNF-α and IL-6 are correlated with al- anine aminotransferase (ALT) in the patients with CH, but not in those with HC with or without asci- tes. Conclusions: These results indicated that MIF, TNF- α and IL-6 may participate in the pathological process of CH and cirrhosis, that IL-6 seems to play an important role in ascites formation, and that se- rum levels of MIF, TNF-α and IL-6 appear to reflect the severity of tissue injury in HBV disease.

Hepatic microenvironment underlies fibrosis in chronic hepatitis B patients

BACKGROUND Chronic hepatitis B virus(HBV)infection is a leading cause of liver morbidity and mortality worldwide.Liver fibrosis resulting from viral infection-associated inflammation and direct liver damage plays an important role in disease management and prognostication.The mechanisms underlying the contribution of the liver microenvironment to fibrosis in HBV patients are not fully understood.There is an absence of effective clinical treatments for liver fibrosis progression;thus,establishing a suitable in vitro microenvironment in order to design novel therapeutics and identify molecular biomarkers to stratify patients is urgently required.AIM To examine a subset of pre-selected microenvironment factors of chronic HBV patients that may underlie fibrosis,with a focus on fibroblast activation.METHODS We examined the gene expression of key microenvironment factors in liver samples from patients with more advanced fibrosis compared with those with less severe fibrosis.We also used the human stellate cell line LX-2 in the in vitro study.Using different recombinant cytokines and growth factors or their combination,we studied how these factors interacted with LX-2 cells and pinpointed the crosstalk between the aforementioned factors and screened the most important factors.RESULTS Of the secreted factors examined,transforming growth factor(TGF)-β1,interleukin(IL)-1βand tumor necrosis factor(TNF)-αwere increased in patients with advanced fibrosis.We found that besides TGF-β1,IL-1βcan also induce a profibrotic cascade by stimulating the expression of connective tissue growth factor and platelet-derived growth factor(PDGF)in LX-2 cells.Furthermore,the proinflammatory response can be elicited in LX-2 cells following treatment with IL-1βand TNF-α,suggesting that stellate cells can respond to proinflammatory stimuli.By combining IL-1βand TGF-β1,we observed not only fibroblast activation as shown byαlpha-smooth muscle actin and PDGF induction,but also the inflammatory response as shown by increased expression of IL-1β.CONCLUSION Collectively,our data from HBV patients and in vitro studies demonstrate that the hepatic microenvironment plays an important role in mediating the crosstalk between profibrotic and proinflammatory responses and modulating fibrosis in chronic HBV patients.For the establishment of a suitable in vitro microenvironment for HBV-induced liver fibrosis,not only TGF-β1 but also IL-1βshould be considered as a necessary environmental factor.

Dysfunction of peripheral blood dendritic cells from patients with chronic hepatitis B virus infection

AIM To identify the property of dendritic cella (DCs) of peripheral blood monocytes (PBMC) in patlents with chronic HBV infection.METHODS Twenty patients with persistent HBV infectlon were included in this study, 10 healthy subjects being used as a control group. The peripheral blood mononuclear cells (PBMC) of T cell-depleted populations were incubated and induced into mature dendritic cells in the RPMI-1640 medium in the presence of cytokines GMCSF, IL-4, FLt-3, TNF-α and 100 mL@ L-1 of fetal calf serum for a total of 10 - 12 days. The expressions of surface markers on DCs were evaluated using flow cytometric analysis. ELISA method was used to determine the cytokine levels of interleukin-12 (IL-12) and IL-10 in the supernatant produced by DCs. For detection of the stimulatory capacity of DCs to T cell proliferation,mytomycin C-treated DC were incubated with allogenic T cells.RESULTS A typical morphology of mature DCs from healthy subjects and HBV-infected patients was induced in in vitro incubation, but the proliferation ability and cellular number of DCs from HBV-infected patients significantly decreased compared with healthy individuals. In particular, the expression levels of HLADR, CD80 (B7-1) and CD86 (B7-2) on DC surface from patients were also lower than that from healthy individuals (0.46 vs 0.92 for HLA-DR, 0.44 vs 0.88 for CD80 and 0.44 vs 0. 84 for CD86, P＜ 0.05). The stimulatory capacity and production of IL-12 of DCs from patients in allogenic mixed lymphocyte reaction (AMLR) significantly decreased, but the production level of nitric oxide (NO) by DCa simultaneously increased compared with healthy subjects (86± 15 vs 170±22 μmoI@L 1, P＜0.05).CONCLUSION The patients with chronic HBV infection have the defective function and immature phenotype of dendritic cells, which may be associated with the inability of efficient presentation of HBV antigens to host immune system for the clearance of HBV.

Risk of hepatitis B virus reactivation in patients with autoimmune diseases undergoing non-tumor necrosis factor-targeted biologics

Hepatitis B virus reactivation(HBVr)can occur in patients treated with immunosuppressive medications.Risk stratification for HBVr based on hepatitis B virus(HBV)serology and viral load is an important strategy to determine appropriate HBV monitoring and antiviral prophylaxis use.Recent advances in the understanding of pathophysiology of autoimmune diseases have led the development of cytokine-targeted therapies.Tumor necrosis factor(TNF)-αinhibitors have been widely used for patients with inflammatory bowel disease,psoriasis,and rheumatic diseases.Further,the clinical benefits of interleukin(IL)-12/23,IL-17,or Janus kinases inhibitors have been demonstrated in these patients.It is well known that TNF-αinhibitor use can lead to HBVr,however,the risk of HBVr in patients undergoing non-TNF-targeted biologics have not been fully understood.In this review,we discuss the risk of HBVr in patients treated with non-TNF-targeted biologics,and immunological mechanisms of these medications causing HBVr.

Controversial issues regarding the roles of IL-10 and IFN-γ in active/inactive chronic hepatitis B

According to the important roles played by cytokines in induction of appropriate immune responses against hepatitis B virus(HBV),Dimitropoulou et al have examined the important cytokines in their patients.They showed that the serum levels of interleukin 10(IL-10)and interferon-γ(IFN-γ)were decreased in patients with HBeAg-negative chronic active hepatitis B compared with the inactive hepatitis B virus carriers(Dimitropoulou et al 2013).The controversy can be considered regarding the decreased serum levels of IFN-γin the HBeAg-negative chronic active hepatitis B patients.They concluded that subsequent to decreased expression of IFN-γ,the process of HBV proliferation led to liver diseases.Previous studies stated that HBV is not directly cytopathic for the infected hepatocytes and immune responses are the main reason for destruction of hepatocytes(Chisari et al,2010).Scientists believe that immune responses against HBV are stronger in active forms of chronic HBV infected patients than inactive forms(Zhang et al,2012).Therefore,the findings from Dimitropoulou et al may deserve further attention and discussion.Additionally,downregulation of IL-10 inchronically active hepatitis B infected patients has also confirmed our claim.IL-10 is an anti-inflammatory cytokine and its expression is increased in inactive forms in order to downregulate immune responses(Arababadi et al,2012).Thus,based on the results from Dimitropoulou et al,it can be concluded that increased immune responses in chronically active hepatitis B infected patients are related to declined expression of IL-10 and interestingly IFN-γis not involved in induction of immune responses in these patients.

Role of Th1/Th2 cells activation in chronic hepatitis B virus infection

Objective: The percentages of Th0, Th1, Th2 cells among single CD4+ T cells isolated from the peripheral blood mononuclear cells (PBMC) of chronically HBV-infected individuals were investigated by determining the expression of interleukin-4 (IL-4) and interferon-γ (IFN-γ), and the role of Th1/Th2 cytokines in chronic HBV-infection was analyzed. Methods: PBMCs from chronically HBV-infected individuals were separated routinely, stimulated by PMA, ionomycin and monensin, and the production of IL-4 and IFN-γ by CD4+ T cells in PBMC was determined by fluorescent activated cell sorting (FACS) analysis. Results: The percentages of IFN-γ-producing T cells and IL-4-producing T cells among the CD4+ cells ranged from 2. 3% to 18. 6% and from 0. 9% to 9.2% respectively in non-infected individuals. The majority of CD4+ T cells from PBMCs in chronically HBV-infected individuals were Th0 cells. The percentage of Th1 cells increased significantly with the hepatic iflammation activity. The percentage of Th1 cells in advanced stage of chronic hepatitis B was higher than that in stable stage of chronic hepatitis B. The percentage of Th2 cells in CD4+ T cells from HBV-infected individuals did not differ significantly, but higher than that from controls Conclusion: The results indicate that Th1 phenotype cytokines were associated with hepatic inflammation activity of chronic hepatitis B and Th2 cells may be associated with the chronicity of HBV infection.

Distribution specificity of polarized populations of T helper cells in patients with chronic hepatitis B virus infection

Objective:To investigatetherolesof thepolarizedpopulationsof T helpercellsisolatedfromtheperipheral bloodmononuclearcells(PBMCs)of individualswithchronichepatitisB virus(HBV)infection.Methods:PBMCsfrom patientswithchronicHBVinfectionwereseparatedroutinely,stimulatedby PMA,ionomycinandmonensin,andthepro-ductionof IL-4,IFN-γandTGF-βby CD4 + T cellswas observedby flowcytometry(FACS).Results:Thepercentagesof theT cellsproducingIFN-γ,IL-4or TGF-βrangedfrom2.3%to18.6%,1.1%to8.7%and0.7%to7.1%respectivelya-mongCD4 + cellsfromnon-infectedindividuals.Themajorityof CD4 + T cellsin PBMCsfromindividualswithchronic HBVinfectionwereTh0cells.Theproportionof Th1cellsinpatientswithactivechronichepatitisB washigherthanthat in patientsat inactivestageof thedisease(P<0.05),indicatinga significantelevationof Th1cellswiththehepaticinflam-mationactivity.Thepercentageof Th2cellsinindividualswithHBVinfectionwashigherthanthatincontrols(P<0.05),butshowedno differencebetweendifferentpatients(P>0.05).Thepercentageof Th3cellswas higherin asymptomatic HBVcarriersthanthatinpatientswithchronichepatitisB andinhealthycontrols(P<0.05).Conclusions:Th1-typecy-tokinesarerelatedwithhepaticinflammationactivityof chronichepatitisB,andTh2cellsmaybe associatedwiththeper-sistenceof HBVinfection.Th3cellscooperatingwithTh2cellsarelikelyto functionas negativeimmunoregulator,and maybe responsiblefortheimmunetolerancestateof chronicHBVinfection.

Gene polymorphisms of interleukin-28, p21-activated protein kinases 4, and response to interferon-α based therapy in Chinese patients with chronic hepatitis B

Background Peg-lnterferon-a treatment is expensive and associated with considerable adverse effects, selection of patients with the highest probability of response is essential for clinical practice. The objective of this study was to assess the relationship between the gene polymorphisms of interleukin-28 （IL-28）, p21-activated protein kinase 4 （PAK4） and the response to interferon treatment in chronic hepatitis B patients. Methods Two hundred and forty interferon-naive treatment HBeAg seropositive chronic hepatitis B patients were enrolled in the present prospective nested case-control study. Peripheral blood samples were collected, including 92 with favorable response and 148 without response to the interferon treatment. Rs8099917, rs12980602, and rs9676717 SNP was genotyped using matrix assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF MS）. Results IL-28 genotype was not associated with response to interferon treatment （OR for GT/GG vs. TT, 0.881 （95% CI 0.388-2.002）; P=0.762; OR for CT/CC vs. TT, 0.902 （95% CI 0.458-1.778）; P=-0.766）. Rs9676717 in PAK4 genotype was independently associated with the response （OR for CT/CC vs. TT, 0.524 （95% CI 0.310-0.888）; P=0.016）. When adjusting for age, gender, smoking, drinking, levels of hepatitis B virus DNA, and alanine aminotransferase （ALT）, rs9676717 genotype TT appeared to be associated with a higher probability of response for interferon treatment （OR, 0.155 （95% CI 0.034-0.700）; P=0.015）. Conclusion Genotype TTfor rs9676717 in PAK4 gene and no drinking may be predictive of the interferon-a treatment success.

白介素⁃15基因多态性与中西医结合治疗HBeAg阴性慢性乙型肝炎患者疗效的关联性分析

目的分析白介素-15(IL-15)基因多态性与中西医结合治疗乙型肝炎病毒e抗原(HBeAg)阴性慢性乙型肝炎(CHB)患者疗效的关联性。方法将312例HBeAg阴性CHB患者随机分为治疗组、对照组,每组156例,两组均给予西医抗病毒治疗,在此基础上,治疗组加用补肾健脾利湿颗粒,对照组加用补肾健脾利湿颗粒安慰剂,疗程为1年。比较两组的治疗应答率,观察血清IL-15水平的变化情况;挑选IL-15基因的标签单核苷酸多态性(SNPs),并进行基因分型,分析IL-15基因多态性与治疗应答的关联性。结果①最终完成试验者306例,其中治疗组155例、对照组151例。②两组治疗应答率比较,差异无统计学意义(P>0.05)。③治疗前后组内比较,治疗组血清HBsAg水平降低、IL-15水平升高(P<0.05),对照组血清HBsAg水平降低(P<0.05);组间治疗后比较,血清HBsAg、IL-15水平差异有统计学意义(P<0.05)。④治疗后,两组治疗应答患者血清IL-15水平升高、无应答患者血清IL-15水平降低(P<0.05),治疗组应答患者血清IL-15水平高于治疗组无应答患者(P<0.05)。⑤关联分析结果显示,有5个SNPs与治疗应答有显著性关联证据,分别为rs6819823、rs2857261、rs10519612、rs10519613、rs1057972(P<0.05)。⑥治疗前,治疗组应答患者rs6819823 TT基因型血清IL-15水平低于GG基因型、rs2857261 AA基因型血清IL-15水平低于GG基因型、rs1057972 AA基因型患者血清IL-15水平低于TT基因型(P<0.05);治疗后,携带rs6819823、rs2857261、rs10519612、rs10519613、rs1057972最小等位基因的纯合子和杂合子患者血清中IL-15水平上升(P<0.05)。结论IL-15基因遗传变异及IL-15表达水平变化与中西医结合治疗HBeAg阴性CHB的临床疗效存在关联。

Plasmacytoid Dendritic Cell Function and Cytokine Network Profiles in Patients with Acute or Chronic Hepatitis B Virus Infection

Background： Plasmacytoid dendritic cells （pDCs） and cytokines play an important role in occurrence and recovery of hepatitis B virus （HBV） infection. The aim of this study was to explore the frequency and function ofpDC and serum cytokine network profiles in patients with acute or chronic HBV infection. Methods： The healthy individuals （HI group）, hepatitis B envelope antigen （HBeAg）-positive chronic HBV patients in immune tolerance （IT） phase （IT group）, HBeAg-positive chronic HBV patients （CHB group）, and acute HBV patients （AHB group） were enrolled in this study. The frequency of cluster of differentiation antigen 86 （CD86） ＋ pDC and the counts of CD86 molecular expressed on surface ofpDC were tested by flow cytometer. The quantitative determinations ofcytokines, including Fins-like tyrosine kinase 3 ligand （FIt-3L）, interferon （1FN）-α2, IFN-γ, interleukin （IL）-17A, IL-6, IL-10, transforming growth factor （TGF）-β1 and TGF-β2, were performed using Luminex multiplex technology. Results： In this study, there were 13 patients in HI group, 30 in IT group, 50 in CHB group, and 32 in AHB group. Compared with HI group, HBV infected group （including all patients in IT, CHB and AHB groups） had significantly higher counts of CD86 molecular expressed on the surface ofpDC （4596.5 ± 896.5 vs. 7097.7 ± 3124.6; P 〈 0.001）. The counts of CD86 molecular expressed on the surface of pDC in CriB group （7739.2 ±4125.4） was significantly higher than that of IT group （6393.4 ± 1653.6, P=0.043）. Compared with IT group, the profile of cytokines of FIt-3L, IFN-γ, and IL-17A was decreased, IFN-α2 was significantly increased （P =0.012） in CH B group. The contents of IL-10, TGF-{31, and TGF-132 in AHB group were significantly increased compared with IT and CHB groups （all P 〈 0.05）. Conclusions： This study demonstrated that the function of pDC was unaffected in HBV infection. The enhanced function of pDC and IFN-α2 might involve triggering the immune response from IT to hepatitis active phase in H BV infection. Acute patients mainly presented as down-regulation of the immune response by enhanced IL-10 and TGF-β.

Impact of Interleukin-28B Polymorphism on Response to Treatment in Chinese Hepatitis C Patients

The candidate polymorphism rs12979860 of interleukin 28B（IL28B） gene was genotyped by means of polymerase chain reaction（PCR） amplification and direct PCR sequencing, and then the role of this single nucleotide polymorphism（SNP） in the response to the treatment of 172 Chinese patients with hepatitis C virus（HCV） chronic infection was analyzed. The results show that the frequencies of major homozygotes（CC） and heterozygotes（CT） of rs12979860 were 0.84 and 0.16, respectively, and the minor homozygotes（TT） wasn’t found in this cohort. A highly significant association was found between the CC genotype and sustained virological response（SVR） in HCV geno type 1 infected patients（P〈0.001） but not in HCV non-genotype 1 infected patients. In addition, a strong association was found between rs12979860 CC genotype and rapid virological response（RVR） in both HCV genotype 1 infected patients（P〈0.001） and non-genotype 1 infected patients（P〈0.001）. Taken together, these results indicate that IL28B polymorphism plays a key role in response to hepatitis C therapy in Chinese patients. Combine assessment of clinical predictors and the IL28B polymorphism may be beneficial to the individualized treatment decision in Asian chronic hepatitis C（CHC） patients.

DETECTION OF PLASMA SOLUBLE INTERLEUKIN－2 RECEPTOR IN PATIENTS WITH SEVERE AND CHRONIC ACTIVE HEPATITIS B

Plasma levels of soluble interleukin-2 receptor (sIL-2R) in patients with chronic active hepatitis B (CAHB) or severe hepatitis B (SHB) were measured quantitatively by 'sandwich' ELISA with monoclonal antibodies in order to explore the change of sIL-2R levels, its clinical significance,and its relation to liver damage. The results showed that the plasma sIL-2R levels in patients with CAHB and SHB were much higher than those in normal controls (P < 0. 01 ), and the level ofplasma sIL-2R in patients with SHB was greatly higher than that in patients with CAHB. These results suggest that there is close relation between plasma level of sIL-2R, the clinical types of hepatitis B,and the severity of liver damage. In addition, there is no significant difference in plasma levels of sIL-2R between acute severe hepatitis B (ASHB), subacute severe hepatitis B (SASHB), and chronic severe hepatitis B (CSHB). No relation was found between sIL-2R level and hepatitis B virusreplication activity.

Emerging Role of Interleukin 22 in Hepatitis B Virus Infection: a Double-edged Sword

Hepatitis B virus (HBV) infection remains a worldwide health problem,and is the major cause of hepatitis,liver cirrhosis,and hepatocellular carcinoma.The innate and adaptive immune responses of the HBV-infected host contribute greatly to the development and pathogenesis of chronic HBV infection,and often affect the efficacy of anti-HBV drugs.Interleukin (IL)-22 is a newly identified cytokine that is involved in the pathogenesis of liver disease,but its role in liver inflammation in patients with HBV infection remains controversial.In this report,we summarize the production and function of IL-22 in inflammatory environments,and review the current research into IL-22 biology in HBV infection.A better understanding of the intrahepatic micro-environments that directly influence the activity of IL-22 will be important for the development of new immunotherapeutic approaches that target IL-22-producing cells or IL-22 itself.

IL28B polymorphism genotyping as predictor of rapid virologic response during interferon plus ribavirin treatment in hepatitis C virus genotype 1 patients

AIM: To clarify the association of interleukin-28B(IL28B) single nucleotide polymorphisms(SNPs) with hepatitis C virus(HCV) viremia changes for assessment of interferon(IFN) response. METHODS: A cohort of 118 Caucasian treatment-nave HCV-G1 infected patients, treated with pegylated-IFN alpha 2a or 2b associated with ribavirin(53 responders, 65 non-responders) during the period 2010-2012, were genotyped for IL28 B SNPs rs12979860 C>T and rs8099917 T>G. Genotyping was performed by realtime allelic discrimination assay. Serum HCV RNA levels were assayed at 2, 4, 12, 24 and 48 wk during therapy.Correlation between IL28 B genotypes and serum HCV RNA kinetics was investigated. Multivariable logistic regression analysis was performed to identify predictors of null-response.RESULTS: Twenty-six out of 118 patients(22%) had no HCV RNA decline ≥ 1 log IU/mL at therapy week 4(null-responders). IL28 B genotype was rs8099917(G*)/rs1297860(**) in 21/26(80%) of null-responder patients. Using multivariate analysis, it was shown that the presence of the rs8099917 G allele was the best predictor of null-response(OR = 7.9, 95%CI: 1.99-31.18). The presence of at least one favorable genotype showed a positive predictive value of above 90% for HCV RNA reduction ≥ log at week 4. Analysis of the HCV RNA kinetics during 12 wk of therapy in patients with IL28 B rs12979860 CT heterozygosis(n = 73), according to their rs8099917 status, showed that the viremia reduction was significantly different in patients carrying the rs8099917 G allele compared to those with favorable homozygosis.CONCLUSION: Our findings emphasize the association of the IL28 B rs8099917 G allele with HCV. Genotyping for both IL28 B SNPs is useful in clinical practice for thorough patient risk stratification based on IFN responsiveness.

Interleukin-10 Is Expressed in HepG2.2.15 Cells and Regulated by STAT1 Pathway

This study investigated the expression profiles of IL-10 gene in three human hepatoma cell lines including Huh7, HepG2, and HepG2 transfected with a plasmid containing hepatitis B virus （HBV） named HepG2.2.15. RT-PCR analysis demonstrated that IL-10 message RNA was absent in HepG2 and Huh7 cells, whereas it was present in HepG2.2.15 cells, which was consistent with ELISA result. Furthermore, except for lamivudine other antiviral treatments did not significantly decrease the HBV DNA level in HepG2.2.15 cells, while they had different effects on the expression of IL-10 protein, although stimulation by LPS had no significant effect. In addition, except for poly（I：C）, the other treatments decreased the expression of IL-10 protein to different degrees, but had no sig-nificant effects on the expression of NF-κB and MyD88. Meanwhile, all treatments we used had effect on the expression of STAT1. In conclusion, IL-10 was expressed in HepG2.2.15 cells and STAT1 pathway might be involved in the regulation of IL-10 expression in HepG2.2.15 cells, but it was not the sole pathway, the exact mechanism warrants further study.

Is the use of IL28B genotype justified in the era of interferon-free treatments for hepatitis C?

In 2009, several groups reported that interleukin-28B(IL28B) genotypes are associated with the response to peginterferon plus ribavirin therapy for chronic hepatitis C virus(HCV) infection in a genome-wide association study, although the mechanism of this association is not yet well understood. However, in recent years, tremendous progress has been made in the treatment of HCV infection. In Japan, some patients infected with HCV have the IL28 B major genotype, which may indicate a favorable response to interferon-including regimens; however, certain patients within this group are also interferon-intolerant or ineligible. In Japan, interferonfree 24-wk regimens of asunaprevir and daclatasvir are now available for HCV genotype 1b-infected patients who are interferon-intolerant or ineligible or previous treatment null-responders. The treatment response to interferon-free regimens appears better, regardless of IL28 B genotype. Maybe other interferon-free regimens will widely be available soon. In conclusion, although some HCV-infected individuals have IL28 B favorable alleles, importance of IL28 B will be reduced with availability of oral interferon free regimen.