Alteration Of Glycans Profiling Of Serum Glycoproteins During Seroconversion Processes In The HBV Bearing Patients Treated With Antivirals Therapy And Its Clinical Significance

ObjectiveTo study the alteration of glycan affinity profiles of serum glycoproteins bylectin microarray during the HBeAg and HBsAg seroconversion in the chronichepatitis B patients after antivial therapy, and to explore its biological significance.For the clinical treatment of chronic hepatitis B, efficacy evaluation, prognosis orfurther understanding of the pathogenesis of chronic hepatitis B provide importantinformationMethodsThe high abundance proteins depleted serum proteins were collected and then were labeled with fluorescence dye Cy5. The lectin microarray containing50lectinswere employed in this assay to detect their affinity to special glycan of proteins.Lectin-affinity glycan profiles of serum protein between control group, HBeAg+non-treated group、HBeAg seroconversion after anti-HBV therapy group and HBsAgloss after anti-HBV therapy group were compared, and the results from lectinmicroarray were partially verified by lectin blot. Experimental data were analyzed byone-way analysis of variance, and afterwards with Student-Newman-keuls (SNK)pairwise comparisons.ResultsBoth lectin microary and lectin bloting assay showed significantly reducedaffinity on16kinds of lectins in HBeAg positive non-treated group compared controlgroup(P<0.05), it showed that the specifc glycan profiles in HBeAg positivenon-treated group may include the decreased terminal and core fucose, GalNAc α-Thr／Ser(T, Tn-antigen), GalNac α, terminal β1-4, and β-D galactose, bisecting and/orGlcNac, mannose and Sia. However, in HBeAg seroconversion after anti-HBVtherapy group,including PSA、MPL and above-mentioned16lectins bind signal ofserum glycoprotein glycan were enhance(P<0.05),which suggested that these reducedserum glycoprotein glycan structures in HBeAg seroconversion after anti-HBVtherapy group were returned to normal or slightly higher than healthy levels. Whencomparing HBsAg loss after anti-HBV therapy group to HBeAg seroconversion afteranti-HBV therapy group, the binding ability of ten lectins, AAL, ACL, HAL, HPL,RCA-I, LEL, STL, PHA-E, NML and PCL were weakened to near control levels;while six lectins, VAL, LCA, GNL, PSA, MPL and JAC had strengthened theirbinding ability to special glycan, there were statistically difference (p<0.05).This implied that the glycan containing terminal fucose, GalNacα, terminal β1-4galactose,bisecting GlcNAc glycan structures dropped to near control levels, while the terminalβ-D-galactose residues and core fucose structure increased significantly.ConclusionSerum glycan of glycoproteins were altered during the HBeAg and HBsAgseroconversion in chronic hepatitis B patients. This may imply they are closely relatedto the seroclerance HBV of CHB, and special lectin binding glycan of proteins mayact as the sugar markers to explore the mechanism of HBsAg seroclearance duringanti-HBV therapy for chronic hepatitis B.Novelty and the potential application of Project1. Our study is the first time to compare the serum glycoprotein profiling duringHBeAg and HBsAg seroconversion processes in the CHB patients treated withantivirals therapy.2. The high-throughput lectin microarray technology and lectin boltting assayrevealed specificity enhanced serum glycoptotein affinity of VAL and LCA inHBsAg loss. And core fucose and terminal β-D-galactose residues structure mayact as the sugar markers associated with the disappearance of serum HBsAgduring anti-HBV therapy for chronic hepatitis B. The potential value of thesespecific glycan anomalous as HBeAg and HBsAg seroconversion processes isworth further studying.