T-bet/GATA-3 ratio as a measure of the Th1/Th2 cytokine profile in mixed cell populations: predominant role of GATA-3

被引:160
作者
Chakir, H
Wang, HP
Lefebvre, DE
Webb, J
Scott, FW
机构
[1] Ottawa Hlth Res Inst, Ottawa, ON K1H 8L6, Canada
[2] Univ Ottawa, Dept Biochem Microbiol & Immunol, Ottawa, ON K1N 6N5, Canada
[3] Univ Ottawa, Dept Med, Ottawa, ON, Canada
基金
加拿大创新基金会; 加拿大健康研究院;
关键词
Th1/Th2; transcription; cytokine; spleen; T cell differentiation; T-bet; GATA-3;
D O I
10.1016/S0022-1759(03)00200-X
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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. (C) 2003 Elsevier B.V. All rights reserved.
引用
收藏
页码:157 / 169
页数:13
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