Yang Min, Wang Shengjun, Wang Suoying, Ma Jie, Xu Xiaopeng, Mao Chaoming, Ma Bing, Tong Jia, Qiu Gufeng, Shao Qixiang, Ding Qing, Xu Huaxi
Department of Immunology, School of Medical Technology, Jiangsu University, Zhenjiang, China.
Immunol Invest. 2008;37(2):97-111. doi: 10.1080/08820130701690725.
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.
T-bet是一种Th1特异性转录因子，可促进γ干扰素的产生。γ干扰素是主要的Th1效应细胞因子，在Th1分化中起关键作用，可驱动初始CD4 + T细胞分化为辅助性T细胞1（Th1）细胞。在我们的研究中，将人T-bet基因与编码HIV-1蛋白转导结构域的基因片段在细菌表达载体中融合，以产生Tat/T-bet融合蛋白。表达并纯化的Tat/T-bet蛋白以时间和剂量依赖性方式有效地转导到THP-1细胞中；当用Tat/T-bet预处理的THP-1细胞与CD4 + T细胞共培养时，在48小时检测时，γ干扰素水平升高至约7 pg/ml，比正常水平高10倍。结果表明，Tat/T-bet融合蛋白可有效地转导到像THP-1细胞这样的抗原呈递细胞（APC）中，进而调节Th1/Th2平衡，这可能作为基因治疗的潜在工具。
