Role of WASP/WIP Complex in T Cell Function

T细胞功能障碍导致Wiskott-Aldrich综合征患者易患 感染、自身免疫和淋巴系统恶性肿瘤。本项目探讨了WASP及其合作伙伴的功能 黄蜂相互作用蛋白WIP在T细胞中。 WASP主要与T细胞中的WIP形成复合体。黄蜂在T细胞中的水平从正常水平下降到~10% WIP KO小鼠和来自AS患者的突变扰乱了WIP结合，表明WIP 稳定WASP。我们的初步数据显示，T细胞功能障碍的等级是WASP/WIP DKOWIP KOWASP KO，请记住，由于WIP KO T细胞中的WASP水平较低，个体 WIP对WIP KO表型的贡献尚不清楚。 我们的总体假设是WIP和WASP在T细胞中既是一个复合体又是一个独立的功能。至 验证我们的假设我们建议使用基因工程小鼠，这些小鼠选择性地缺乏WIP、WASP或 两者，或者其中WASP/WIP复合体被破坏。我们会： 1.比较WASP/WIP DKO小鼠和WT小鼠的T细胞，以确定WIP的联合作用 和WASP.此外，与WASP KO T细胞的比较将定义WIP功能，其可能是 独立于WASP。 2.用含有WASP结合域(WBD)的多肽重建WIP KO小鼠的T细胞 以恢复WASP水平，从而产生WIP缺陷/WASP表达的T细胞。我们会 将这些小鼠的T细胞与WT T细胞进行比较，以确定WIP的作用。 3.破坏WT T细胞中表达WBD的WASP/WIP复合体，以验证存在 依赖于WASP/WIP复合体的信令功能子集。 4.检测表达WIP突变体的敲击小鼠的T细胞的结构和功能，该突变体缺乏 肌动蛋白结合区确定WIP与肌动蛋白结合在亚细胞定位和功能中的作用 WASP/WIP综合体。 在小鼠身上进行的实验结果将有助于我们理解糖尿病的发病机制。 是在人类身上发生的，应该会导致这种疾病的新疗法。

T cell dysfunction contributes to the susceptibility of patients with the Wiskott-Aldrich syndrome (WAS) to infection, autoimmunity and lymphoid malignancy. This project probes the function of WASP and its partner WASP interacting protein WIP in T cells. Most of WASP is complexed with WIP in T cells. WASP levels of drop to ~10% of normal in T cells from WIP KO mice and from WAS patients with mutations that disrupt WIP binding, suggesting that WIP stabilizes WASP. Our Preliminary data show that the hierarchy of T cell dysfunction is WASP/WIP DKOWIP KOWASP KO, bearing in mind that since WASP levels are low in WIP KO T cells the individual contribution of WIP to the WIP KO phenotype is unknown. Our overall hypothesis is that WIP and WASP function in T cells both as a complex and independently. To test our hypothesis we propose to use genetically engineered mice which selectively lack WIP, WASP or both, or in which the WASP/WIP complex is disrupted. We will: 1. Compare T cells from WASP/WIP DKO mice to T cells from WT mice to define the combined role of WIP and WASP. In addition, comparison with WASP KO T cells would define WIP functions that may be independent of WASP. 2. Reconstitute T cells from WIP KO mice with a peptide that consists of the WASP binding domain (WBD) of WIP in order to restore WASP levels, and thus generate WIP deficient/WASP expressing T cells. We will compare T cells from these mice to WT T cells to define the role of WIP. 3. Disrupt the WASP/WIP complex expressing the WBD in WT T cells to test the hypothesis that there is a subset of signaling functions that is dependent on the WASP/WIP complex. 4. Examine the structure and function of T cells from knockin mice that express a WIP mutant that lacks the actin-binding region to define the role of WIP binding to actin in the subcellular localization and function of the WASP/WIP complex. The results of the proposed experiments in the mouse will aid in our understanding of the pathogenesis of WAS in humans and should lead to novel treatments for this disease.