Macrophage migration inhibitory factor is increased in the urine of patients with urinary tract infection: Macrophage migration inhibitory factor-protein complexes in human urine
Macrophage migration inhibitory factor is increased in the urine of patients with urinary tract infection: Macrophage migration inhibitory factor-protein complexes in human urine
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DOI：
10.1016/s0022-5347(05)00650-6
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发表时间：
2006-04-01
影响因子：
6.6
通讯作者：
Vera, PL
中科院分区：
文献类型：
--
作者：
Meyer-Siegler, KL;Iczkowski, KA;Vera, PL
目的：MIF 是一种促炎细胞因子，存在于人尿路上皮的预先形成的储存中。在膀胱炎症（包括细菌性膀胱炎）的动物模型中，MIF 在膀胱中上调，并以高分子量复合物的形式从膀胱释放。我们比较了 UTI 患者和无 UTI 患者的尿液 MIF 含量，并检查和鉴定了尿液中的 MIF-蛋白复合物。 材料和方法：使用酶联免疫吸附测定，我们比较了 14 名 UTI 患者和 16 名无 UTI 对照者尿液中的 MIF 水平。在天然、变性和还原条件下进行蛋白质印迹以检查尿液中发现的 MIF 复合物。质谱法鉴定出尿液中的 MIF 相关蛋白，而共免疫沉淀证实了这种关联。结果：通过酶联免疫吸附测定测定的 14 名 UTI 患者的平均尿液 MIF 含量 SEM 显着高于 16 名对照者（1.96 +/- 0.40 vs 0.59 +/- 0.09 ng/mg 肌酐，p < 0.01）。变性条件下的蛋白质印迹显示，UTI 尿液中的几种高分子量复合物（100 至 165 kDa）以及典型的单体 MIF（12 kDa）增加。质谱法鉴定了相关的 MIF 蛋白，包括铜蓝蛋白、白蛋白和尿调节蛋白。免疫共沉淀证实了质谱分析结果，并确定了 MIF 与 α-2-巨球蛋白的相互作用。结论：细菌性膀胱炎患者尿液中 MIF 含量增加支持了我们的实验证据，表明 MIF 在盆腔内脏炎症中发挥作用。 MIF 与其他尿蛋白关联的新发现表明，MIF 的生理相关形式可能是 MIF-蛋白复合物。
Purpose: MIF is a proinflammatory cytokine present in preformed stores in human urothelium. In animal Models of bladder inflammation, including bacterial cystitis, MIF is up-regulated in the bladder and released from the bladder as a high molecular weight complex. We compared urine MIF amounts in patients with UTI to that in patients without UTI, and we examined and identified MIF-protein complexes in urine.Materials and Methods: Using enzyme-linked immunosorbent assay we compared MIF levels in the urine of 14 patients with UTI to levels in 16 controls with no UTI. Western blotting under native, denaturing and reducing conditions was done to examine MIF complexes found in urine. Mass spectrometry identified MIF associated proteins in urine, while coimmunoprecipitation confirmed the associations.Results: Mean urine MIF amounts SEM determined by enzyme-linked immunosorbent assay were significantly greater in 14 patients with UTI compared to that in 16 controls (1.96 +/- 0.40 vs 0.59 +/- 0.09 ng/mg creatinine, p < 0.01). Western blotting under denaturing conditions showed several high molecular weight complexes (100 to 165 kDa) that increased in UTI urine as well as typical, monomeric MIF (12 kDa). Mass spectrometry identified associated MIF proteins, including ceruloplasmin, albumin and uromodulin. Co-immunoprecipitation confirmed mass spectrometry findings and also identified MIF interaction with alpha-2-macroglobulin.Conclusions: Increased urine MIF amounts in patients with bacterial cystitis support our experimental evidence showing a role for MIF in pelvic visceral inflammation. The novel finding of an association of MIF with other urine proteins suggest that the physiologically relevant form of MIF may be an MIF-protein complex.