Le Bras Severine, Massaad Michel, Koduru Suresh, Kumar Lalit, Oyoshi Michiko K, Hartwig John, Geha Raif S
Division of Immunology, Children's Hospital, and Department of Pediatrics, Harvard Medical School, Boston, MA 02115, USA.
Proc Natl Acad Sci U S A. 2009 May 5;106(18):7519-24. doi: 10.1073/pnas.0806410106. Epub 2009 Apr 9.
The Wiskott-Aldrich syndrome (WAS) interacting protein (WIP) stabilizes the WAS protein (WASP), the product of the gene mutated in WAS. WIP-deficient T cells have low WASP levels, limiting the usefulness of WIP KO mice in defining the role of WIP in T cell function. To define this role, we compared WIP/WASP double KO (DKO) mice to WASP KO mice on DO11.10 background. T cell development was normal in both strains, but peripheral T cell numbers were significantly decreased in DKO mice. WASP KO T cells proliferated and secreted IL-2 normally in response to OVA peptide (OVAp). In contrast, T cells from DKO mice proliferated poorly in response to OVAp in vitro, and cutaneous hapten hypersensitivity was deficient in these mice. DKO T cells up-regulated CD25 expression and secreted normal amounts of IL-2 after antigen stimulation, but had defective response to IL-2, evidenced by failure to further up-regulate CD25 expression, phosphorylate STAT5, and induce expression of STAT5-dependent genes. DKO, but not WASP KO, T cells had a disrupted subcortical actin cytoskeleton and impaired actin polymerization after T cell antigen receptor (TCR) ligation. These results indicate that WIP is essential for IL-2 signaling and responsiveness in T cells, possibly because of its critical role in TCR-triggered actin cytoskeletal reorganization.
威斯科特-奥尔德里奇综合征(WAS)相互作用蛋白(WIP)可稳定WAS蛋白(WASP),WASP是WAS中发生突变的基因的产物。缺乏WIP的T细胞中WASP水平较低,这限制了WIP基因敲除(KO)小鼠在确定WIP在T细胞功能中作用方面的用途。为了明确这一作用,我们将DO11.10背景下的WIP/WASP双基因敲除(DKO)小鼠与WASP基因敲除小鼠进行了比较。两种品系的T细胞发育均正常,但DKO小鼠的外周T细胞数量显著减少。WASP基因敲除的T细胞对卵清蛋白肽(OVA p)有正常的增殖反应并分泌白细胞介素-2(IL-2)。相比之下,DKO小鼠的T细胞在体外对OVA p的增殖反应较差,并且这些小鼠缺乏皮肤半抗原超敏反应。DKO T细胞在抗原刺激后上调了CD25表达并分泌了正常量的IL-2,但对IL-2的反应存在缺陷,表现为未能进一步上调CD25表达、磷酸化信号转导和转录激活因子5(STAT5)以及诱导STAT5依赖性基因的表达。DKO而非WASP基因敲除的T细胞在T细胞抗原受体(TCR)连接后,其皮质下肌动蛋白细胞骨架受到破坏且肌动蛋白聚合受损。这些结果表明,WIP对于T细胞中的IL-2信号传导和反应性至关重要,这可能是因为它在TCR触发的肌动蛋白细胞骨架重组中起关键作用。