The Relationship Between Serum CD4~+T Lymphocyte Associated Cytokines And The Efficacy Of Antiviral Therapy In Hbeag Positive Chronic Hepatitis B

Objective: Chronic hepatitis B (CHB) is a worldwide epidemic disease，with a high incidence it may evolve to cirrhosis, liver failure andhepatocellular carcinoma (HCC), which are major threats to human health.Knowledge of natural history of CHB indicates that continuous replication ofhepatitis B virus (HBV) is an independent risk factor of progression of CHB.Thus, antiviral treatment is the key to the treatment of CHB. Currently,interferon (Pegylated or conventional interferon alpha, PEG-IFN or IFNα) andnucleos(t)ide analogues (lamivudine, adefovir, telbivudine, entecavir, etc) aretwo major types of drugs that can be used in the treatment of CHB, they cannot only effectively inhibite HBV replication, but also can alleviate the illnessand prevent progression of the disease to cirrhosis, liver failure, HCC and soon. However, none of the drugs above can eliminate the HBV effectivelybecause of the complex interaction between HBV, host immune status andliver condition, therefore, it is difficult to achieve the goal of clinical cure.Host immune response plays a pivotal role in the recognition and eliminationof HBV, which may also be an important factor affecting antivirol efficacy andclinical outcomes. Meanwhile, the effector CD4+T cell-mediated specificimmune response which is an important part of immune system are alsoinvolved. Previous studies have indicated that HBV-specific CD4+T celldisfunction is one of the causes of chronic HBV infection, the resultingimmune dysfunction may also associated with treatment response to antiviraltherapy in CHB. However, these studies have focused more on detecting theCD4+T cell directly, although reflected the frequency of the cell moreaccurately, but the detection method was more complicated and costly. Thedifferentiation, function and regulation of CD4+T cell subsets are achieved through a cytokine network, a variety of cytokines are included. Therefore,detect the CD4+T lymphocyte associated cytokines can not only reflect itsfunctional status, the method is more simple and low cost, which is conductiveto clinical applications. In the current study, we dynamically observed changesof CD4+T lymphocyte associated cytokines levels during different antiviralregimens in patients with CHB, in order to map the relationship between CD4+T lymphocyte associated cytokines and the efficacy of antiviral therapy inHBeAg positive CHB more precisely and provide a basis for guiding clinicaltreatment.Methods:111HBeAg positive CHB patients were recruited from theThird Hospital of Hebei Medical University, the Fifth Hospital ofShijiazhuang city and the Sixth Hospital of Handan city between February2012and January2014. Among them,60cases were treated with entecavir0.5mg/d orally for48weeks, the other51cases were terated with Pegylated IFNα2a180μg or135μg weekly or Pegylated IFNα2b1.5μg/kg (weight) weeklyfor48weeks. To assess the antivirol efficacy, we defined three responsegroups according to HBVDNA and HBeAg by treatment week48,(i.ecomplete response was defined as HBVDNA undetectable and HBeAg loss;partial response was defined as a decrease in HBVDNA of≥2lgcopies/ml,HBeAg positive; non-response was defined as a decrease in HBVDNA of＜2lgcopies/ml, HBeAg positive). Serum samples were collected at baseline(0W) and12W，24W and48W after treatment. Liver and renal functionparameters, serum virology and HBV DNA loads were determined at the sametime. Serum CD4+T lymphocyte associated cytokines concentrations weredetermined by enzyme-linked immunosorbent assays (ELISA). Compare theirlevels before and after treatment in different response group，and dynamicallyobserve their changes during therapy. Comparison among groups andcorrelation analysis were performed by SPSS13.0.Results:1Baseline demographic and clinical characteristics of patients and theresponse to different treatment of all patients. Among the51patients treated with PEG-IFNα,18(35.29%) achieved acomplete response (CR), and the baseline HBVDNA concentration of patientswho achieved a CR was significantly lower than that of non-responders(P=0.036).13(21.67%) of60patients treated with ETV were achieved a CRand their baseline ALT levels were singnificantly higher than that ofnon-responders (P=0.025).2Serum CD4+T cell subsets associated cytokines changes duringPEG-IFNα treatment and their effect on antiviral efficacy.Among our patients treated with PEG-IFNα，IP10，IL13，IL18and IL21showed significant change in different response groups，no other cytokineswere associated with the antiviral outcome.Baseline serum IP10concentration was significantly higher in CR groupthan in PR group (P=0.048) and NR group (P＜0.001). There was a decreaseof serum IP10concentration after12weeks treatment in all response groups,but the decreasing level was significantly higher in CR group than in PR groupand NR group (P＜0.05).There was no significant difference of serum IL13concentration in allresponse groups (P＞0.05) before treatment, while in patients who achieved aCR or PR showed a significant decrease in IL13than non-responders by week12(P＜0.05).There was no significant difference of serum IL18concentration in anytwo response groups in all the observation points, while we observed asignificant increase in patients who achieved a CR or PR by week24, whichwas not shown in NR group.There was no significant difference of baseline serum IL21concentrationin any two response groups (P＞0.05), while we observed a significantincrease in patients who achieved a CR or PR(P＜0.001) by week12, whichwas significantly higher in CR group than that in PR group(P=0.004) and NRgroup(P=0.001).Univariate logistic analysis showed that baseline HBVDNAconcentration＜6.45lgcopies/ml (P=0.011), ALT level＞179.5U/L (P=0.015), IP10concentration＞154.245pg/ml (P=0.002), a decrease in IP10＞59.59pg/ml (P＜0.001) by week12, a decrease in IL13＞6.11pg/ml (P=0.011)by week12, an increase in IL18＞26.345pg/ml (P=0.015) by week24and anincrease in IL21＞102.035pg/ml (P=0.001) by week12were associated with aCR. While multivariate analysis showed that only an increase in IL21＞102.035pg/ml (P=0.045) by week12was a predictive factor related to a CR(PPV81.25%, NPV85.71%, AUC0.823).3Serum CD4+T cell subsets associated cytokines changes during ETVtreatment and their effect on antiviral efficacy.Among our patients treated with ETV, IP10and IL21showed significantchange in different response groups, no other cytokines were associated withthe antiviral outcome.Baseline serum IP10concentration was significantly higher in CR groupthan in PR group (P=0.037) and NR group (P＜0.014). There was a decreaseof serum IP10concentration after12weeks treatment in CR and PR groups (P＜0.001), which was not shown in NR group. When the decreasing levelswere compared，we observed a significant difference in any two groups(P＜0.014).There was no significant difference of serum IL21concentration in anytwo response groups (P＞0.05) before treatment, while we observed asignificant increase in patients who achieved a CR or PR(P＜0.001) by week24, which was significantly higher in CR group than that in PR group and NRgroup(P＜0.05).Univariate logistic analysis showed that age of the patients＜38.5yearsold (P=0.015), baseline HBVDNA concentration＜6.99lgcopies/ml (P=0.081),ALT level＞163.5U/L (P=0.003), IP10concentration＞207.69pg/ml(P=0.041)，a decrease in IP10＞71.625pg/ml(P＜0.001) by week12and anincrease in IL21＞75.11pg/ml (P=0.003) by week24were associated with aCR. While multivariate analysis showed that a decrease in IP10＞71.625pg/ml(P=0.015, PPV55.56%, NPV92.86%, AUC0.809) by week12, an increase inIL21＞75.11pg/ml (P=0.035) by week24(PPV44.44%, NPV96.97%, AUC0.797), age of the patients＜38.5years old (P=0.047) and baselineHBVDNA concentration＜6.99lgcopies/ml (P=0.069) were predictive factorsrelated to a CR.4Linear correlation of CD4+T cell subsets associated cytokines withALT and HBVDNA before treatment.Serum IP10(r=0.675, P＜0.001) and IL18(r=0.532, P＜0.001) werepositively correlated with ALT.Serum IL13was positively correlated withHBVDNA load（r=0.271, P=0.004）. No other cytokines showed a correlationwith these two parameters.Conclusions:1.Serum CD4+T cell subsets associated cytokines IP10, IL13and IL18may influence the efficacy of PFGIFNα therapy in HBeAg positive CHB tosome extent, and an increase in serum IL21after12weeks treatment was afactor related to a complete resonse by week48.2. Baseline HBVDNA load, age, a decrease in serum IP10after12weekstreatment and an increase in serum IL21concentration after24weekstreatment were factors related to a complete response by week48in patientstreated with ETV.3. Serum IP10and IL18were positively with ALT，which may indicate arole of IP10and IL18in the immunopathologic damage due to HBV infection.While serum IL13was positively with HBVDNA load before treatment andthis may indicate a relationship between IL13and HBV replication.