Lentivirally overexpressed T-bet regulates T-helper cell lineage commitment in chronic hepatitis B patients

Mol Med Rep. 2012 Aug;6(2):361-6. doi: 10.3892/mmr.2012.905. Epub 2012 May 8.

Abstract

Chronic hepatitis B virus (HBV) infection is commonly considered to occur as a result of disturbance of the immune system. T-box expressed in T cells (T-bet) is an essential transcription factor for T helper (Th) cell differentiation and function. The aim of this study was to investigate the effect of T-bet overexpression on Th cell differentiation and the possible mechanism in chronic hepatitis B (CHB) patients. CD4+ T cells from the peripheral blood of 23 CHB patients, 8 acute hepatitis B (AHB) patients and 10 healthy controls were isolated. T-bet mRNA expression of CD4+ T cells was detected by quantitative real-time polymerase chain reaction (PCR). The T-bet DNA fragment was subcloned into the pGC-FU vector containing GFP to generate a recombinant lentiviral vector, pGC-FU-T-bet, while a no-load pGC-FU vector was used as the negative control. After transduction into CD4+ T cells from another 22 CHB patients, the induction of Th1- and Th2-type cytokines was assayed by an enzyme-linked immunosorbent assay (ELISA), and RT-PCR and western blot analysis were used to measure the mRNA and transcription levels of H2.0-like homeobox (HLX1), GATA-3 and STAT-6. T-bet mRNA expression in CD4+ T cells from AHB patients was enhanced compared with CHB patients and healthy controls. Th1-type cytokines and HLX1 expression was upregulated, while Th2-type cytokines and GATA-3 and STAT-6 expression was repressed after lentiviral introduction of T-bet. In conclusion, lentivirally overexpressed T-bet regulates Th cell lineage commitment in CHB patients, which may be mediated by regulating HLX1, GATA-3 and STAT-6 expression.

Keywords: chronic hepatitis B; T-bet; T helper cells; H2.0-like homeobox; GATA-3; STAT-6.

MeSH terms

  • Adult
  • Blotting, Western
  • Case-Control Studies
  • Cell Differentiation
  • Cell Lineage
  • Cloning, Molecular
  • Cytokines / genetics
  • Cytokines / immunology
  • Cytokines / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • GATA3 Transcription Factor / genetics
  • GATA3 Transcription Factor / metabolism
  • Gene Expression Regulation
  • Genetic Vectors / genetics
  • Genetic Vectors / metabolism
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Hepatitis B virus / immunology
  • Hepatitis B virus / metabolism
  • Hepatitis B virus / pathogenicity
  • Hepatitis B, Chronic / immunology*
  • Hepatitis B, Chronic / metabolism
  • Hepatitis B, Chronic / virology
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism
  • Humans
  • Lentivirus / genetics
  • Lentivirus / metabolism*
  • Male
  • Middle Aged
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Real-Time Polymerase Chain Reaction
  • STAT6 Transcription Factor / genetics
  • STAT6 Transcription Factor / metabolism
  • T-Box Domain Proteins / genetics
  • T-Box Domain Proteins / metabolism*
  • T-Lymphocytes, Helper-Inducer / immunology*
  • T-Lymphocytes, Helper-Inducer / metabolism
  • T-Lymphocytes, Helper-Inducer / virology
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Transcription, Genetic

Substances

  • Cytokines
  • GATA3 Transcription Factor
  • GATA3 protein, human
  • HLX protein, human
  • Homeodomain Proteins
  • RNA, Messenger
  • STAT6 Transcription Factor
  • STAT6 protein, human
  • T-Box Domain Proteins
  • T-box transcription factor TBX21
  • Transcription Factors
  • Green Fluorescent Proteins