The Research On Host Immune Response In Patients With Chronic Hepatitis B During Antiviral Treatment With Nucleoside (Acid) Analogue

Obiective: To investigate the levels of circulating T, B, NK cells and serumcytokines IL-2, IFN, TNF, IL-4, IL-6, IL-10, and analyze the association betweenthese laboratory parameters and liver function and viral loads in patients with chronichepatitis B (CHB), which treated to antiviral treatment, to explore its role in theimmune response.Methods: This study were collected outpatients and hospitalized patients withCHB of the First Hospital of Jilin University, and began screening test after signinginformed consent, we selected54previously untreated HBeAg-positive CHB patientsthat giving them telbivudine,30previously adefovir-resistant (SAR) HBeAg-positiveCHB patients that switching to ETV monotherapy and20gender-, age-, andethnic-matched healthy subjects. The numbers of different subsets of peripheralCD3-CD56+, CD244+NK, CD3+, CD3+CD4+, CD3+CD8+, CD4+CD25+Foxp3+,CD4+CD25+CD127lowT, CD19+CD27+B cells and the frequency ofcytokine-secreting CD4+T cells (IL-2, IFN, TNF, IL-4, IL-6, IL-10) in subjectswere measured by flow cytometry analysis and micro-array flow cytometry (CBA).The levels of serum ALT and AST were detected using a Biochemistry AutomaticAnalyzer (Roche Diagnostics). HBV-related HBsAg, HBsAb, HBeAg, and HBeAbwere detected by a chemiluminescent microparticle immunoassay (CMIA) using anAbbott I2000automated chemiluminescence immunoassay analyzer (AbbottLaboratories). The amount of serum HBV DNA and HCV RNA were measured byquantitative PCR assay using a luciferase quantization detection kit, following theprotocols. The potential association of the laboratory parameters and clinical measureswas also analyzed.Results: Previously untreated HBeAg-positive CHB patients that giving themtelbivudine:1. Following treatment with LDT, the levels of HBV DNA loads and thelevels of serum HBsAg, ALT and AST in the CHB patients gradually decreased andwere significantly lower at3,6,9,13months post initial treatment than those before treatment. There were5,8,12,15cases with seronegative HBeAg at3,6,9,13months post initial treatment.2. Following treatment with LDT, the numbers ofCD3+CD8+T, CD3-CD56+, CD244+NK, and cytokine-secreting CD4+T cells (IL-2,IFN, TNF, IL-4, IL-6, IL-10) were increased, but CD3+CD4+,CD4+CD25+Foxp3+, CD4+CD25+CD127low, CD8+PD-1+T cells in CHB patientswere decreased as compared with that before treatment (P<0.001) and was graduallyelevated at the later time points to a level similar to that in the HC.3. The frequencyof cytokine-secreting CD4+T cells was negatively associated with the levels of serumHBV DNA loads, ALT and AST in CHB patients.Previously adefovir-resistant (SRA) HBeAg-positive CHB patients thatswitching to ETV monotherapy:1. After switching to ETV monotherapy, the levels ofHBV DNA loads and the levels of serum HBsAg, ALT and AST in the SRA CHBpatients gradually decreased and were significantly lower at3,6months than thosebefore treatment. There were4,11, cases with seronegative HBeAg at3,6, monthsafter switching to ETV monotherapy.2. After switching to ETV monotherapy, thenumbers of CD3+CD8+T, CD3-CD56+, CD244+NK, and cytokine-secreting CD4+T cells (IL-2, IFN, TNF, IL-4, IL-6, IL-10) were increased, but CD3+CD4+,CD4+CD25+Foxp3+, CD4+CD25+CD127lowT and CD19+CD27+B cells in SRACHB patients were decreased as compared with that before treatment (P<0.001).3.After switching to ETV monotherapy, the frequency of CD4+IFN+T cells wasnegatively associated with the levels of serum HBV DNA loads, ALT and AST inSRA CHB patients.Conclusions:1. The natural immunity and acquired immunity in patients withchronic B hepatitis were flawed.2. Changes of immune cells and its associatedcytokines in the antiviral treatment process suggest that immunomodulatory mightplay a role in the inhibition of viral after application of nucleos (t) ide analogues.3.SRA CHB patients to switch to ETV monotherapy to continue antiviral treatmentmight get better treatment effect, suggesting that ETVmonotherapy could be usedadopted as a rescue therapy for suppression of SRA HBV strains.4. Virologicalresponse analysis showed the association between the levels of cytokine-secretingCD4+T cells and HBV DNA, suggesting that serum cytokines were closely related tothe viral replication in the process of antiviral therapy.5. The levels ofcytokine-secreting CD4+T cells could be used as clinical indicators for monitoring the course of early stages of antiviral treatment, to evaluate prognosis and the diseaseoutcome in CHB patients.