The Antiviral Treatment Of Hepatitis B Patients TCR Variable Region Deep Sequencing Analysis

BackgroundAccording to the World Health Organization reported that about20million people worldwidehave been infected with HBV, Of which350million people are chronically infected, Every year,1millionpeople who are chronically infected with HBV die of liver failure, liver cirrhosis, or hepatocellularcarcinoma.Our contury is a high incidence of chronic hepatitis B prevalence area, Chronic HBV infection isapproximately93million, There needed to be treated the symptoms of hepatitis B patients with active isabout2000million. The antiviral efficacy of patients with chronic hepatitis B virus itself and the influenceof such factors as host, but the host cell immune state is one of the important factors in the therapeuticresponse and plays an important role in scavenging liver damage and viral cells. In the acute self-limitedHBV infection, the cellular immune response against HBV relatively strong, however, in chronic hepatitisB, the immune response against HBV is much weaker. Virus infection mediated by immune cells to causeliver damage, T cell dysfunction may be the root cause of chronic HBV infection. The current treatment ofchronic hepatitis B main measures are: hepatoprotective drugs, anti-viral and immune modulation therapy,it can not completely eradicate HBV. In the course of the immune response to HBV, the specific cytotoxicT cells (CTL) determines the outcome of the disease and has played an important role in clearing the virus.T cell receptor (,TCR) determines the CTL specificity which is the feature of variable area V area mainlydepends on the molecular composition. To research the composition of TCR features can help to deepunderstanding of HBV reaction mechanism of the immune response.In the recent years, with the rapid development of next-generation sequencing technology andthe completion of the Human Genome Project, High-throughput sequencing technology with highthroughput, simple operating platform, quality advantages has been applied in various fields of medicine.This experiment uses the before and after the peripheral blood lymphocytes in patients with chronichepatitis B amplification receptor ?-chain variable region of high-throughput sequencing and analysis ofchanges in the types and the CDR clone in the receptor area and help to clarify the characteristics of the mechanism of cellular immune responses in patients with CHB.To reveal the pathogenesis of chronichepatitis B, effective treatment strategies to provide new ideas and technology platforms experimentalstudies and Provide assistance to immunotherapy of chronic hepatitis B.PurposesUse the Ion Torrent high-throughput sequencing technology to observe the treatment of chronichepatitis B patients differentially expressed after TCR ? chain V region gene in family,to delve into thepathogenesis of chronic hepatitis B patients to lay the experimental foundation solution HBV reactionmechanism of the immune response.Materials and methods?1?the study object: Collected from March2010to December2013in Beijing youan hospitalaffiliated to the capital university of medical sciences, clinical treatment of outpatient service effectiveHBeAg positive chronic hepatitis B patients, all patients were accord with the2010edition of the chronichepatitis B prevention guide diagnostic criteria. Merging other virus infectionand other causes of liverdisease patients, the treatment of chronic hepatitis B is not suitable for antiviral treatment.(2) methods: collect blood samples of CHB and peripheral blood mononuclear cell, after theextraction of total RNA extraction mRNA,cDNA was synthesis, CDR3region amplification,CDR3sizeanalysis, high-throughput sequencing was performed according to the manufacturer's instructions.Results: After the transition from raw database,there are about380000clean reads, considering V,J family genes at the same time.All30V family and J family genes to build database,and do the Blast andfind15genes were upregulated, the most significant two genes were up-regulated most significant, therewas statistically significantConclusions:The.abnormal.expression.of.many.genes,such.asTRBV10_TRBJ2.1,TRBV10_TRBJ2.2, TRBV28_TRBJ1.5?TRBV6_TRBJ2.1and so on,were found in CD8+T cell of patients withHBV.They may involve in epathogenesis of HBV. It is possible that TRBV28_TRBJ1.5, TRBV6_TRBJ2.1may take part in immune reponse.