Search for a novel eosinophil activation biomarker and risk assessment for asthma using the biomarker

寻找新型嗜酸性粒细胞激活生物标志物并使用该生物标志物进行哮喘风险评估

基本信息

批准号：
18590550
负责人：
KAMBAYASHI Yasuhiro
金额：
$ 2.32万
依托单位：
Kanazawa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

The specificity of the rabbit antiserum raised against brominated keyhole limpet hemocyanin was investigated with an enzyme-linked immunosorbent assay (ELISA). The antiserum recognized brominated bovine serum albumin (BSA), chlorinated BSA, dibromotyrosine-conjugated BSA, and di-iodetyrosine-conjugated BSA However, the antiserum did not recognize bromotyrosine-conjugated BSA and chlorotyrosine-conjugated BSA Moreover, the specificity of the antiserum was investigated with competitive ELISA Dibromotyrosine and di-iodetyrosine inhibited the recognition of bromintaed BSA by the antiserum. These results suggested that the epitope of the antiserum is dihalogentated tyrosine in protein. Immunohistochemically, the antiserum stained brominated rat eosinophlis but not chlorinated eosinophils. It was suggested that the antiserum could recognize dibromotyrosine in protein as eosinophil activation biomarker in immunohistochmical analysis.The antiserum cannot be used as specific antiserum for dibomotyrosine, eosinophil activation biomarker, in ELISA, because the antiserum recognized dihalogenated tyrosine in protein. However, both dibromotyrosine and dichlorotyrosine were formed in allergic disease such as asthma. Thus, severity of asthma may be judged using the halogenated tyrosine in protein as a biomarker. The antiserum recognized brominated BSA in the presence of human plasma. After the antiserum was preincubated with human plasma and various dose of brominated BSA for 2hr, dose-dependent inhibition of the recognition of antigen by the antiserum. These results showed that the antiserum could be used for the measurement of dihalogenated tyrosine in protein of human plasma.It is expected that the severity of asthma is judged using competitive ELISA with the antiserum. Immunohistochemical analysis with the antiserum could be used to elucidate the pathophysiology of asthma in mouse asthma model.

采用酶联免疫吸附测定 (ELISA) 研究了针对溴化匙孔血蓝蛋白的兔抗血清的特异性。抗血清识别溴化牛血清白蛋白(BSA)、氯化BSA、二溴酪氨酸结合BSA和二碘酪氨酸结合BSA，但是，该抗血清不识别溴酪氨酸结合BSA和氯酪氨酸结合BSA。此外，该抗血清的特异性为通过竞争性 ELISA 研究二溴酪氨酸和二碘酪氨酸抑制识别抗血清对溴化 BSA 的影响。这些结果表明，抗血清的表位是蛋白质中的二卤化酪氨酸。免疫组织化学显示，抗血清对溴化大鼠嗜酸性粒细胞染色，但对氯代嗜酸性粒细胞不染色。提示该抗血清在免疫组织化学分析中可以识别蛋白质中的二溴酪氨酸作为嗜酸性粒细胞活化生物标志物。该抗血清不能用作ELISA中嗜酸性粒细胞活化生物标志物二溴酪氨酸的特异性抗血清，因为该抗血清识别蛋白质中的二卤代酪氨酸。然而，二溴酪氨酸和二氯酪氨酸都是在哮喘等过敏性疾病中形成的。因此，可以使用蛋白质中的卤化酪氨酸作为生物标志物来判断哮喘的严重程度。在人血浆存在的情况下，抗血清可识别溴化 BSA。抗血清与人血浆和不同剂量的溴化BSA预孵育2小时后，抗血清对抗原的识别呈剂量依赖性抑制。这些结果表明，该抗血清可用于人血浆蛋白中二卤代酪氨酸的测定。期望用该抗血清进行竞争性ELISA来判断哮喘的严重程度。抗血清的免疫组织化学分析可用于阐明小鼠哮喘模型中哮喘的病理生理学。