Inoue Keiichi, Wang Xintao, Saito Junpei, Tanino Yoshinori, Ishida Takashi, Iwaki Daisuke, Fujita Teizo, Kimura Shioko, Munakata Mitsuru
Department of Pulmonary Medicine, School of Medicine, Fukushima Medical University, Fukushima, Japan.
Allergol Int. 2008 Mar;57(1):57-64. doi: 10.2332/allergolint.O-07-493. Epub 2008 Mar 1.
Uteroglobin-related protein 1 (UGRP1) is a secretory protein expressed in the airways and is speculated to have anti-inflammatory activity. In the mouse, its gene expression is down-regulated by interleukin (IL)-5 and -9, and up-regulated by IL-10. However, the precise role of UGRP1 in human inflammatory airway diseases such as asthma has not been clarified. The objectives of this study were to establish an ELISA system to quantify UGRP1 protein, and to examine whether plasma UGRP1 levels are associated with the G-112A polymorphism, asthma susceptibility, and its severity.
152 asthma patients and 103 normal controls were involved in this study. Mice were immunized with recombinant UGRP1 and hybridoma cell lines were established. A sandwich ELISA system was established by using two monoclonal antibodies recognizing different epitopes. Plasma UGRP1 levels were measured with the ELISA system and the G-112A allele was detected by using real-time PCR.
An ELISA system was established that allowed determination of UGRP1 levels within the range of 9.6-1250 pg/ml. The mean plasma UGRP1 levels for subjects with -112A allele were significantly lower than those without it (p = 0.025). Although there was no significant difference in the plasma UGRP1 levels between asthma patients and controls (p = 0.13), severe asthma patients without oral corticosteroid had significantly lower plasma UGRP1 levels compared to mild- or moderate- asthma patients and controls (p = 0.004, 0.03 and 0.003, respectively).
The ELISA system for quantifying UGRP1 protein was established, and plasma UGRP1 levels were associated with the G-112A UGRP1 gene promoter polymorphism and the severity of asthma.
子宫珠蛋白相关蛋白1(UGRP1)是一种在气道中表达的分泌蛋白,据推测具有抗炎活性。在小鼠中,其基因表达受白细胞介素(IL)-5和-9下调,受IL-10上调。然而,UGRP1在人类炎性气道疾病如哮喘中的具体作用尚未阐明。本研究的目的是建立一种酶联免疫吸附测定(ELISA)系统来定量UGRP1蛋白,并检测血浆UGRP1水平是否与G-112A多态性、哮喘易感性及其严重程度相关。
本研究纳入了152例哮喘患者和103名正常对照。用重组UGRP1免疫小鼠并建立杂交瘤细胞系。使用两种识别不同表位的单克隆抗体建立夹心ELISA系统。用ELISA系统测量血浆UGRP1水平,并通过实时聚合酶链反应(PCR)检测G-112A等位基因。
建立了一种ELISA系统,可在9.6 - 1250 pg/ml范围内测定UGRP1水平。携带-112A等位基因的受试者血浆UGRP1平均水平显著低于未携带该等位基因的受试者(p = 0.025)。尽管哮喘患者和对照之间的血浆UGRP1水平无显著差异(p = 0.13),但未使用口服糖皮质激素的重度哮喘患者与轻度或中度哮喘患者及对照相比,血浆UGRP1水平显著降低(分别为p = 0.004、0.03和0.003)。
建立了定量UGRP1蛋白的ELISA系统,血浆UGRP1水平与UGRP1基因启动子G-112A多态性及哮喘严重程度相关。