Tat-mediated intracellular delivery of T-bet protein into THP-1 cells can induce Th1-type response

Immunol Invest. 2008;37(2):97-111. doi: 10.1080/08820130701690725.

Abstract

T-bet, a Th1-specific transcription factor, can promote the production of IFN-gamma. IFN-gamma is the principal Th1 effector cytokine and it has a crucial role in Th1 differentiation, which can drive the differentiation of naïve CD4+T cells into T-helper 1 (Th1) cells. In our study, a human T-bet gene was fused with a gene fragment encoding HIV-1 protein transduction domain in a bacterial expression vector to produce a Tat/T-bet fusion protein. The expressed and purified Tat/T-bet proteins were transduced efficiently into THP-1 cells in a time- and dose-dependent manner; when Tat/T-bet pretreated THP-1 cells were co-cultured with CD4+T cells, the IFN-gamma level increased higher to about 7 pg/ml, 10-folds as compared with the normal level when tested at 48 hours. The results demonstrated that the Tat/T-bet fusion protein can be efficiently transduced into antigen-presenting cells (APCs) like THP-1 cells and then regulated Th1/Th2 balance, which may act as a potential tool for gene therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigen-Presenting Cells / immunology
  • Blotting, Western
  • Cell Differentiation / immunology
  • Cell Line
  • Enzyme-Linked Immunosorbent Assay
  • Flow Cytometry
  • Genes, tat / immunology*
  • Genetic Therapy / methods*
  • Genetic Vectors
  • HIV / genetics*
  • Humans
  • Interferon-gamma / biosynthesis
  • Lymphocyte Activation / immunology
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology*
  • Reverse Transcriptase Polymerase Chain Reaction
  • T-Box Domain Proteins / genetics*
  • T-Box Domain Proteins / immunology
  • Th1 Cells / cytology
  • Th1 Cells / immunology*
  • Th2 Cells / cytology
  • Th2 Cells / immunology
  • Transduction, Genetic

Substances

  • Recombinant Fusion Proteins
  • T-Box Domain Proteins
  • T-box transcription factor TBX21
  • Interferon-gamma