Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110.
J Immunol. 2013 Dec 15;191(12):5933-40. doi: 10.4049/jimmunol.1300454. Epub 2013 Nov 11.
Plasmacytoid dendritic cells (pDCs) are a dendritic cell subset that secrete type I IFNs in response to microbial stimuli. The scaffold protein, CD2-associated protein (CD2AP), is a marker of human pDCs as it is highly expressed in this cell type. Recently, in human pDCs, decreased CD2AP expression appeared to enhance the production of type I IFNs via an inhibitory receptor-induced signaling cascade. In this study, we sought to determine the role of CD2AP in murine pDCs using CD2AP knockout (KO) mice. CD2AP was dispensable for the development of pDCs and for the upregulation of activation markers following stimulation. Loss of CD2AP expression did not affect the production of type I IFNs stimulated by TLR ligation, and only slightly impaired type I IFN production when inhibitory pathways were engaged in vitro. This was also confirmed by showing that CD2AP deficiency did not influence type I IFN production by pDCs in vivo. Because CD2AP plays a role in regulating actin dynamics, we examined the actin cytoskeleton in pDCs and found that activated CD2AP KO pDCs had significantly higher levels of actin polymerization than wild-type pDCs. Using two different inflammation models, we found that CD2AP KO pDCs have a defect in lymph node migration, correlating with the defects in actin dynamics. Our work excludes a role for CD2AP in the regulation of type I IFNs in pDCs, and suggests that the major function of CD2AP is on the actin cytoskeleton, affecting migration to local lymph nodes under conditions of inflammation.
浆细胞样树突状细胞 (pDCs) 是一种树突状细胞亚群,它在受到微生物刺激时会分泌 I 型干扰素。支架蛋白 CD2 相关蛋白 (CD2AP) 是人类 pDCs 的标志物,因为它在这种细胞类型中高度表达。最近,在人类 pDCs 中,CD2AP 表达的减少似乎通过抑制性受体诱导的信号级联增强了 I 型干扰素的产生。在这项研究中,我们使用 CD2AP 敲除 (KO) 小鼠来确定 CD2AP 在小鼠 pDCs 中的作用。CD2AP 对于 pDCs 的发育以及刺激后激活标记物的上调是可有可无的。CD2AP 表达的丧失并不影响 TLR 连接刺激下产生的 I 型 IFNs,并且仅在体外抑制途径被激活时略微损害 I 型 IFN 的产生。通过显示 CD2AP 缺陷不影响体内 pDCs 产生 I 型 IFN 也证实了这一点。由于 CD2AP 在调节肌动蛋白动力学中起作用,我们检查了 pDCs 中的肌动蛋白细胞骨架,发现激活的 CD2AP KO pDCs 的肌动蛋白聚合水平明显高于野生型 pDCs。使用两种不同的炎症模型,我们发现 CD2AP KO pDCs 在淋巴结迁移中存在缺陷,这与肌动蛋白动力学缺陷相关。我们的工作排除了 CD2AP 在 pDCs 中调节 I 型 IFNs 的作用,并表明 CD2AP 的主要功能是在肌动蛋白细胞骨架上,在炎症条件下影响向局部淋巴结的迁移。