Immunological Characteristics Of Different Clinical Outcomes Of Hbeag Serology In Children With Chronic Hbv Infection And Establishment Of Gender And Age-Specific Reference Intervals For Serum Alt,Ast,Akp And Ggt In Children And Its Application In The Nat

PART 1.IMMUNOLOGICAL CHARACTERISTICS OF DIFFERENT CLINICAL OUTCOMES OF HBEAG SEROLOGY IN CHILDREN WITH CHRONIC HBV INFECTIONObjective: To analyze the immunological characteristics of different clinical outcomes of HBe Ag serology in children with HBe Ag-positive chronic hepatitis B,and to screen key immunological indicators related to HBe Ag seroconversion to distinguish different clinical outcomes about HBe Ag.It provides a new perspective for the follow-up monitoring and treatment effect judgment of children with chronic hepatitis B.Methods: Fifteen children with HBe Ag-positive chronic hepatitis B were enrolled,with an average follow-up of 2.8 years(range: 0.7-6.2 years).At the end of the study,children were divided into three groups according to the status of HBe Ag: spontaneous HBe Ag clearance/seroconversion group(Group1,n=5),persistent HBe Ag positive group(Group2,n=5),and antiviral drug-induced HBe Ag seroconversion group(Group3,n=5).The serum was collected for liver function,hepatitis B markers and HBV-DNA load,and semi-quantitative microarray was used to detect 34 cytokine expression profiles such as IFN-γ、TNF-α、IL-2、IL-4、IL-6、TGF-β、IL-10、IL-17、IL-22 and IL-21.By comparing the expression levels of related cytokines at different time points in each group,the differences and dynamic changes between the groups were compared and analyzed.Boolean models were used to investigate the relationship between cytokines and different clinical outcomes of HBe Ag,and to screen key immunological markers that can distinguish different clinical outcomes of HBe Ag.Results: The overall level of cytokine expression patterns in the spontaneous HBe Ag serological clearance/conversion group(Group1)was low.Antiviral drug-induced HBe Ag seroconversion group(Group3)had the highest level of cytokine expression,and cytokine expression levels remained high after HBe Ag seroconversion during antiviral therapy.At baseline,the expression levels of IL-1b,IL-6R,IL-17 F,IL-21 R,MIP-3α and IL-17 R,IL-22,IL-28 A and GITR in Group3 were significantly higher than those in the other two groups.At the end of follow-up,the expression levels of IL-1b,IL-6R,IL-23p19,IL-28 A and MIP-3α in Group3 were significantly higher than those in the other two groups.The expression levels of IL-1s RI,IL-1s RII,IL-4,IL-12p40,IL-17 F,GITR and TGF-β3 in the persistent HBe Ag-positive group(Group2)increased significantly with time.The expression levels of GITR,IL-22,IL-21 R and IL-17 R at baseline can better distinguish the different clinical outcomes of HBe Ag.In addition,a simple procedure for determining the different clinical outcomes of HBe Ag in children with chronic hepatitis B was established through the expression of baseline GITR and IL-21 R.Conclusion: The expression levels of cytokines in HBe Ag-positive chronic hepatitis B children are quite different with different clinical outcomes.The overall pattern of cytokine expression can better reflect their immune characteristics.By combining the expression levels of GITR,IL-22,IL-21 R,and IL-17 R at baseline,it is possible to better distinguish chronic hepatitis B children with different clinical outcomes about HBe Ag.A simple procedure based on the expression of baseline GITR and IL-21 R was established to determine the clinical outcome of serum HBe Ag in children with HBe Ag-positive chronic hepatitis B.PART 2 ESTABLISHMENT OF GENDER AND AGE-SPECIFIC REFERENCE INTERVALS FOR SERUM ALT,AST,AKP AND GGT IN CHILDREN AND ITS APPLICATION IN THE NATURAL HISTORY OF CHRONIC HBV INFECTIONObjective: Establish gender and age-specific reference intervals for children’s serum ALT,AST,AKP and GGT,and re-evaluate the natural history of chronic hepatitis B in children using the new upper limit of normal(ULN).Methods: Information on the age,sex and liver chemistry of healthy children aged 1-14 was collected from the hospital electronic database system.The age-and sex-specific reference intervals for serum ALT,AST,AKP,and GGT were determined using percentiles method(3rd percentile-97 th percentile).The sensitivity and specificity were calculated to determine the ability of the newly proposed ALT ULN to detect children with chronic HBV infection.Results: The age-and sex-specific RIs of ALT,AST,AKP and GGT were established based on 4232 healthy children.Using the new median ALT threshold,the sensitivity was higher.The detection of chronic HBV infection was 31.2% in boys and 35.5% in girls,while a very slight decrease in specificity was found.Much the same trends of the liver chemistry of CHB children was found relative to healthy children across all sexes and age categories,but still have some differences.Based on the newly proposed RIs of ALT,approximately 16.1% boys and 19.0% girls would be classified in the HBe Ag positive chronic hepatitis phase,but using the current ALT threshold of children’s hospitals they were in HBe Ag positive chronic infection phase.Conclusions: Based on a large healthy population,we established the sex-and age-specific RIs of ALT,AST,AKP and GGT serum levels for children.Meanwhile,we described the feature of liver chemistry in CHB children and newly proposed liver chemistry RIs will benefit the understanding of liver function and the natural history of chronic HBV infection in children.