T-bet/GATA-3 ratio as a measure of the Th1/Th2 cytokine profile in mixed cell populations: predominant role of GATA-3.
The differentiation of naive T-helper (Th) cells towards Th1 or Th2 cells is regulated by the transcription factors T-box expressed in T-cells (T-bet) and GATA-binding protein-3 (GATA-3). In the present study, the gene expression of T-bet and GATA-3 was measured by semi-quantitive reverse transcription polymerase chain reaction (RT-PCR) in Th1 and Th2 cells derived from purified splenic CD4+ T cells from DO11.10/Rag2(-/-) transgenic mice and control BioBreeding (BBc) Wistar rat splenic T cells stimulated under Th1 or Th2 conditions. In both sets of experiments, changes in the ratio of expression of T-bet and GATA-3 reflected changes in the Th1-specific cytokine interferon-gamma (IFN-gamma) and Th2-specific cytokine interleukin (IL)-4. T-bet gene expression was not maintained in fully polarized rat Th1 cells whereas GATA-3 gene expression was maintained in long-term polarized rat Th2 cells, indicating that maintenance of Th1/Th2 status occurred more as a result of altered GATA-3 mRNA expression than T-bet. These transcription factors are up-regulated in several cells that produce type 1 and type 2 cytokines and can be analyzed readily by RT-PCR using total RNA isolated from mixed cell populations or cultured splenocytes thereby providing a surrogate marker of Th1/Th2 cytokine balance under a variety of conditions.
Chakir
Habiba
H
Ottawa Health Research Institute, Ottawa, ON, Canada.
Wang
Haiping
H
Lefebvre
David E
DE
Webb
John
J
Scott
Fraser W
FW
D000818:Animals; D016207:Cytokines / Q000032:analysis / Q000276:immunology*; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D050990:GATA3 Transcription Factor; D015870:Gene Expression; D005786:Gene Expression Regulation; D008213:Lymphocyte Activation; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020825:T-Box Domain Proteins; D018417:Th1 Cells / Q000276:immunology*; D018418:Th2 Cells / Q000276:immunology*; D015534:Trans-Activators / Q000096:biosynthesis* / Q000235:genetics; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D015854:Up-Regulation; D000098815:T-bet Transcription Factor
D016207:Cytokines; D004268:DNA-Binding Proteins; D050990:GATA3 Transcription Factor; C494748:Gata3 protein, mouse; D020825:T-Box Domain Proteins; D000098815:T-bet Transcription Factor; D015534:Trans-Activators; D014157:Transcription Factors