Substance P increases cell-surface expression of CD74 (receptor for macrophage migration inhibitory factor): in vivo biotinylation of urothelial cell-surface proteins.
Substance P increases cell-surface expression of CD74 (receptor for macrophage migration inhibitory factor): in vivo biotinylation of urothelial cell-surface proteins.
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DOI：
10.1155/2009/535348
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发表时间：
2009
影响因子：
4.6
通讯作者：
Vera PL
中科院分区：
文献类型：
--
作者：
Meyer-Siegler KL;Xia SL;Vera PL
巨噬细胞移动抑制因子(MIF)是一种炎性细胞因子，其受体CD74在膀胱炎症中表达上调。MIF介导的信号转导涉及与细胞表面CD74的结合，本研究在体内证明了MIF-CD74在尿路上皮细胞表面的相互作用。生理盐水和P物质(SP，40μg/kg)处理大鼠的N-羟基磺基琥珀酰亚胺生物素酯标记的表面尿路上皮蛋白。用组织学和共聚焦显微镜检查膀胱。生物素化蛋白用亲和素琼脂糖凝胶纯化，用抗MIF或抗CD74抗体免疫沉淀，用链霉亲和素-辣根过氧化物酶检测。只有浅表的尿路上皮细胞被生物素标记。这些细胞含有生物素标记的MIF/CD74细胞表面复合体，这种复合体在SP处理的动物中增加。SP处理可增加尿路上皮细胞MIF和CD74mRNA的表达。我们的数据表明，腔内MIF，作为SP治疗的结果，从尿路上皮细胞释放，与尿路上皮细胞表面CD74相互作用。这些结果证明我们之前描述的MIF-CD74相互作用发生在尿路上皮细胞表面。
Macrophage migration inhibitory factor (MIF), an inflammatory cytokine, and its receptor CD74 are upregulated by bladder inflammation. MIF-mediated signal transduction involves binding to cell-surface CD74, this study documents, in vivo, MIF-CD74 interactions at the urothelial cell surface. N-hydroxysulfosuccinimide biotin ester-labeled surface urothelial proteins in rats treated either with saline or substance P (SP, 40 μg/kg). The bladder was examined by histology and confocal microscopy. Biotinylated proteins were purified by avidin agarose, immunoprecipitated with anti-MIF or anti-CD74 antibodies, and detected with strepavidin-HRP. Only superficial urothelial cells were biotinylated. These cells contained a biotinylated MIF/CD74 cell-surface complex that was increased in SP-treated animals. SP treatment increased MIF and CD74 mRNA in urothelial cells. Our data indicate that intraluminal MIF, released from urothelial cells as a consequence of SP treatment, interacts with urothelial cell-surface CD74. These results document that our previously described MIF-CD74 interaction occurs at the urothelial cell surface.
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