Abnormal IL-4 gene expression by atopic dermatitis T lymphocytes is reflected in altered nuclear protein interactions with IL-4 transcriptional regulatory element
Abnormal IL-4 gene expression by atopic dermatitis T lymphocytes is reflected in altered nuclear protein interactions with IL-4 transcriptional regulatory element
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DOI：
10.1111/1523-1747.ep12340181
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发表时间：
1996-05-01
影响因子：
6.5
通讯作者：
Hanifin, JM
中科院分区：
文献类型：
--
作者：
Chan, SC;Brown, MA;Hanifin, JM
在特应性疾病中，特应性皮炎的特征是血清IgE水平最高，外周血T细胞白介素4(IL-4)产生增加。IL-4促进B细胞合成IgE，刺激产生IL-4的T细胞生长，可能参与了本病的发病机制。在这项研究中，原位杂交证实，与正常受试者相比，特应性皮炎患者外周血中产生IL-4的T细胞频率更高。这些体内来源的T细胞被用来检测产生IL-4的信号要求，以及与IL-4转录调控元件(相对于IL-4转录起始部位-激活反应元件)-88至-60之间的关键转录调控元件相关的核因子。我们证明，在T细胞系中，属于转录因子NF-AT和AP-1家族的蛋白质存在于受刺激的细胞提取物中，并与激活反应元件特异地相关。IL-4的异常产生反映在与该元素相关的核蛋白中。使用凝胶漂移分析，我们发现12个来自刺激的特应性T细胞的核提取物中有12个形成了激活依赖的蛋白质-DNA复合体，而12个正常的T细胞提取物中只有2个形成了激活依赖的蛋白质-DNA复合体。激活复合体的形成与刺激T细胞产生的IL-4mRNA和蛋白的相对水平相关，提示特应性疾病中IL-4基因的异常表达可能与核蛋白与这些启动子元件相互作用的改变有关。
Among the atopic diseases, atopic dermatitis is characterized by the highest levels of serum IgE and by increased peripheral blood T-cell interleukin-4 (IL-4) production. IL-4 promotes IgE synthesis by B cells and stimulates the growth of IL-4-producing T cells and may contribute to the pathogenesis of this disease. In this study, in situ hybridization established that atopic dermatitis patients have a higher frequency of IL-4-producing peripheral blood T cell when compared to normal subjects. These in vivo derived T cells were used to examine the signaling requirements for IL-4 production and the nuclear factors that associate with a critical IL-4 transcriptional regulatory element between -88 and -60 relative to the IL-4 transcription initiation site, the activation responsive element. We demonstrate that, as in T-cell lines, proteins belonging to the NF-AT and AP-1 family of transcription factors are present in stimulated cell extracts and specifically associate with the activation responsive element. Dysregulated IL-4 production is reflected in the nuclear proteins that associate with this element. Using gel shift assays, we found that 12 of 12 nuclear extracts from stimulated atopic T cells formed the activation-dependent protein-DNA complex, compared to only 2 of 12 normal T-cell extracts. Activation complex formation correlated with the relative level of IL-4 mRNA and protein produced in stimulated T cells, suggesting that abnormal IL-4 gene expression in atopic disease may be linked to alterations in nuclear protein interactions with these promoter elements.