Immune Repertoire As The Biomarker To Predict The Clinical Outcomes Of Chronic Hepatitis B

ObjectiveThe role of cellular immunity which is characterized by T cell response in HBV infection is important and complex.They can not only lead to the cure of the disease by clear the virus,but also injure the liver cells to develop the chronic hepatitis or even hepatocellular cacinoma.The number of the TCR repertoire is huge and the condition of the whole system can reflect the history of the antigen exposure and the regulation of the immune system.HBsAg is a vital important index to confirm the complete clearance of the infection.Thus,there is a great significance to research the HBsAg clearance related immune repertoire.Fortunately,there is a convenient and efficient method to characterize the immune repertoire.our research is aimed to :1.To characterize the HBV chronic infection related CD8+ TCR repertoire;2.To find the character of the HBsAg clearance related CD8+ TCR repertoire so as to predict and asses the the outcomes of clinical therapy.Methods1.the CHB patients and healthy control is chosen to collect their peripheral blood to get the lymphocytes by density gradient centrifugation.After 76 week treatment,61 patients and 40 healthy control were selected,and their corresponding lymphocytes were picked up for total RNA extraction.Then the total RNA was reverse transcribed to cDNA,and the CDR3 sequence of TCR? was amplified,which was compared with IMGT database then analyzed.2.the CHB patients and healthy control is chosen to collect their peripheral blood to get the lymphocytes by density gradient centrifugation.After 76 week treatment,the patients with or without clearance of HBsAg were selected,and their corresponding lymphocytes were picked up for total RNA extraction.Then the total RNA was reverse transcribed to cDNA,and the CDR3 sequence of TCR? was amplified,which was compared with IMGT database then analyzed.The patients are further classified to groups with autoclearance or IFN-induced clearance to analyze the difference.To find the mechanisms,we assessed the function of the antigen specific T cells and the gene expression of the total T cells.Results1.The reads index of HBV infection group(CHB)is higher than the health control(HC);the diversity of the CHB group is lower than HC group;although the CDR3 length is even,some cases in CHB group present the skewed distribution;the insertion and deletion of genes as well as the usage or the pair of the V and J gene have no difference.Moreover,we find some HBV infection related clonetypes.2.There is no difference in the CDR3 length,the insertion and deletion of genes,size of ndn length as well as the usage or the pair of the V and J gene.However,the HBsAg clearance patients' CDR3 length is normally distributed while the HBsAg no clearance patients' is skewed distributed.All index of the autoclearance group and IFN-induced clearance group is same.So the function of antigen specific T cells are.But the T cell activation and effecter associated gene expression of HBs Ag clearance group is higher than the other group.Conclusion1.The patients with HBV chronic infection receives the standing antigen stimulation,their T cells are highly proliferated.There do existed HBV infection related clonetypes and some of them are widely distributed.2.The proliferation of T cells in patients with HBsAg clearance is weaker and it may be the results of the powerful immune response at the vary early stage.The present method can not tell the difference of the different styles of HBsAg clearance.There may be some other mechanisms involved so the function of antigen specific T cells that comes from those groups is no difference.However,taking total T cells into account,the T cells in HBsAg clearance group is more activated.