Role of WASP/WIP Complex in T Cell Function

T细胞功能障碍导致Wiskott-Aldrich综合征（WAS）患者对以下疾病的易感性： 感染、自身免疫和淋巴恶性肿瘤。本项目探讨了WASP及其合作伙伴的功能 WASP相互作用蛋白在T细胞中的表达。 大多数WASP与T细胞中的CD 4+细胞复合。WASP水平下降到正常T细胞的约10%， 从WAS患者中分离出的突变破坏了WAS的结合，这表明WAS基因的表达可能与WAS基因的表达有关。 稳定WASP。我们的初步数据表明，T细胞功能障碍的层次是WASP/CD 45。 DKOWASPKO，记住，由于WASP水平在DKOWASPKO T细胞中较低，个体 β-内酰胺酶对β-内酰胺酶KO表型的贡献是未知的。 我们的总体假设是，WASP和WASP在T细胞中既作为复合物又独立地起作用。到 为了验证我们的假设，我们建议使用基因工程小鼠，这些小鼠选择性地缺乏WASP，WASP或 两者，或其中WASP/WASP复合物被破坏。我们将： 1.比较来自WASP/CD 3DKO小鼠的T细胞与来自WT小鼠的T细胞，以确定CD 3DKO的联合作用。 和WASP。此外，与WASP KO T细胞的比较将定义可能与WASP KO T细胞相关的免疫功能。 独立于WASP。 2.用由WASP结合结构域（WBD）组成的肽重建来自WARKO小鼠的T细胞 以恢复WASP水平，从而产生WASP缺陷/WASP表达T细胞。我们将 将这些小鼠的T细胞与野生型T细胞进行比较，以确定T细胞的作用。 3.在WT T细胞中破坏表达WBD的WASP/WASP复合物，以检验存在WBD的假设。 依赖于WASP/WASP复合物的信令功能的子集。 4.检查来自表达缺乏T细胞的突变体的敲入小鼠的T细胞的结构和功能。 肌动蛋白结合区，以确定肌动蛋白结合的作用，在亚细胞定位和功能， WASP/WASP复合体。 在小鼠中的拟议实验的结果将有助于我们理解的发病机制， 是在人类，并应导致新的治疗这种疾病。

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.