CXCL4 is a novel nickel-binding protein and augments nickel allergy
CXCL4 is a novel nickel-binding protein and augments nickel allergy
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DOI：
10.1111/cea.12926
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发表时间：
2017-08-01
影响因子：
6.1
通讯作者：
Sugawara, S.
中科院分区：
文献类型：
--
作者：
Kuroishi, T.;Bando, K.;Sugawara, S.
背景：镍（Ni）是最常见的金属过敏原，可诱导TH 1依赖性IV型过敏。虽然Ni 2+被认为是结合到内源性蛋白质，但目前仍不清楚这些Ni结合蛋白是否参与体内Ni过敏。我们先前报道了脂多糖（LPS）在镍过敏小鼠模型中的佐剂作用。由于LPS可诱导多种炎症介质的产生，因此我们推测LPS也可诱导Ni结合蛋白的产生。目的：从LPS注射小鼠血清中纯化并鉴定Ni结合蛋白（称为LPS血清），并在体内模型中检查这些Ni结合蛋白对Ni变态反应的增强作用。BALB/cA小鼠腹腔注射NiCl 2和LPS致敏。致敏后10天，用NiCl 2通过i.d.注射到耳鼻咽部。镍结合蛋白（s）通过镍亲和柱层析和凝胶过滤纯化。结果：脂多糖血清，而不是从盐水注射的小鼠血清，增强镍过敏性炎症引起的耳肿胀。镍结合，但不是非结合级分，从LPS血清纯化增强镍过敏性炎症。质谱和蛋白质印迹法检测到活性组分中的CXCL 4。用Ni-琼脂糖凝胶和表面等离子体共振分析的批分析揭示了CXCL 4和Ni 2+之间的直接结合。重组CXCL 4增强镍过敏性炎症和发挥佐剂作用的敏化phase.Conclusions：这些结果表明，CXCL 4是一种新的镍结合蛋白，增强镍过敏的启发和敏化阶段。这是第一个研究表明，镍结合蛋白增强镍过敏在体内。
Background: Nickel (Ni) is the most frequent metal allergen and induces a TH1-dependent type-IV allergy. Although Ni2+ is considered to bind to endogenous proteins, it currently remains unclear whether these Ni-binding proteins are involved in Ni allergy in vivo. We previously reported the adjuvant effects of lipopolysaccharide (LPS) in a Ni allergy mouse model. As LPS induces a number of inflammatory mediators, we hypothesized that Ni-binding protein(s) are also induced by LPS.Objective: The objective of this study was to purify and identify Ni-binding protein(s) from serum taken from LPS-injected mice (referred as LPS serum) and examined the augmenting effects of these Ni-binding protein(s) on Ni allergy in an in vivo model.Methods: BALB/cA mice were sensitized with an i.p. injection of NiCl2 and LPS. Ten days after sensitization, mice were challenged with NiCl2 by an i.d. injection into ear pinnae. Ni-binding protein(s) were purified by Ni-affinity column chromatography and gel filtration.Results: Lipopolysaccharide serum, but not serum taken from saline-injected mice, augmented ear swelling induced by Ni-allergic inflammation. Ni-binding, but not non-binding fraction, purified from LPS serum augmented Ni-allergic inflammation. Mass spectrometry and Western blotting detected CXCL4 in the active fraction. A batch analysis with Ni-sepharose and a surface plasmon resonance analysis revealed direct binding between CXCL4 and Ni2+. Recombinant CXCL4 augmented Ni-allergic inflammation and exerted adjuvant effects at the sensitization phase.Conclusions: These results indicate that CXCL4 is a novel Ni-binding protein that augments Ni allergy at the elicitation and sensitization phases. This is the first study to demonstrate that the Ni-binding protein augments Ni allergy in vivo.