Investigation of regulation of Th1 cytokine production by Clara cell 10-kDa protein in granulomatous lung diseases

Clara 细胞 10-kDa 蛋白在肉芽肿性肺病中调节 Th1 细胞因子产生的研究

• 批准号: 13670610
• 负责人: YAMADA Gen
• 金额: $ 2.3万
• 依托单位: Sapporo Medical University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670610/
• 关键词: CC10; Th1 cytokine; sarcoidosis; secretoglobin; CC1O

Clara cell 10-kDa protein (CC10) is the predominant product from non-ciliated bronchiolar epithelial cells (Clara cells). Human CC10 is encoded by a three kilo-base (kb) single-copy gene, which contains three exons and two introns and is localized in the long arm of chromosome 11 (11q12.3-q13.1) CC10 is a homodimer of polypeptide of 70 amino acids covalently bound in an anti-parallel manner. CC10 has been proven to be identical with uteroglobin and protein 1. CC10 possesses aniti-inflmamatory function and inhibits interferon-γ function. CC10 forms one of intra- and intercellular regulators involved in inflammation and malignant transformation in the respiratory fields.We investigated single nucleotide polymorphisms (SNPs) of CC10 gene, showing 6 different SNPs. We analyzed guanine-adenine substitution at position 38 (G38A) downstream from the transcription initiation site in patients with sarcoidosis by allele specific PCR and compared clinical parameters and CC10 levels in serum and bronchoalveolar lavage (BAL) fluid among three genotypes. The A allele frequency in sarcoidosis was significantly higher than that of healthy controls. There were no significant differences of gender, ages, distributions of radiogiaphic stages, number of affected organs, ACE activities, or data of BAL examinations among the genotype groups. However, the A/A genotype group had significant greater rates of disease progression compared with the G/G and G/A genotype groups. The A/A genotype group had significantly lower CC10 levels in serum and BAL fluid compared with the G/G and G/A genotype groups. In the reporter gene assay, significantly lower luciferase reporter activities were detected for the 38A construct compared with the 38G construct in the presence of interferon-γ, but not in the absence of interferon-γ. CC10 G38A SNP may influence CC10 protein expression and be associated with disease progression of sarcoidosis.

Clara 细胞 10-kDa 蛋白 (CC10) 是非纤毛细支气管上皮细胞（Clara 细胞）的主要产物。人CC10由3kb单拷贝基因编码，包含3个外显子和2个内含子，定位于11号染色体长臂(11q12.3-q13.1)。CC10是由70个氨基酸以反平行方式共价结合的多肽同二聚体。 CC10已被证明与子宫珠蛋白和蛋白1相同。CC10具有抗炎功能和抑制干扰素-γ功能。 CC10 形成参与呼吸领域炎症和恶性转化的细胞内和细胞间调节因子之一。我们研究了 CC10 基因的单核苷酸多态性 (SNP)，显示了 6 个不同的 SNP。我们通过等位基因特异性 PCR 分析了结节病患者转录起始位点下游 38 位 (G38A) 的鸟嘌呤-腺嘌呤取代，并比较了三种基因型的临床参数以及血清和支气管肺泡灌洗 (BAL) 液中的 CC10 水平。结节病中的 A 等位基因频率显着高于健康对照。各基因型组间性别、年龄、X线分期分布、受累器官数量、ACE活性或BAL检查数据均无显着差异。然而，与 G/G 和 G/A 基因型组相比，A/A 基因型组的疾病进展率显着更高。与G/G和G/A基因型组相比，A/A基因型组血清和BAL液中CC10水平显着降低。在报告基因测定中，在存在干扰素-γ的情况下，检测到38A构建体的荧光素酶报告活性显着低于38G构建体，但在干扰素-γ不存在的情况下则不然。 CC10 G38A SNP 可能影响 CC10 蛋白表达并与结节病的疾病进展相关。

项目成果

Shijubo N, et al.: "Immunological aspects of Clara cell 10-kda protein"Annual Review Immunology. 113-119 (2003)

Shijubo N 等人：“Clara 细胞 10-kda 蛋白的免疫学方面”年度免疫学评论。

Shijubo N et al.: "Mapping of functional epitopes of osteopontin by monoclonal antibodies raised against defined internal sequences"J Cell Biochem. 84・2. 420-432 (2002)

Shijubo N 等人：“通过针对确定的内部序列产生的单克隆抗体绘制骨桥蛋白的功能表位”J Cell Biochem 84·2 (2002)。

Shijubo N, Yamada G et al.: "Interleukin-18 in inflammatory lung diseases"Recent Res Devel Resp Critical Care Med. 223-231 (2001)

Shijubo N、Yamada G 等人：“炎症性肺部疾病中的白介素 18”Recent Res Devel Resp Critical Care Med。

四十坊典晴, 山田玄ほか: "肺の炎症の免疫学的解析法"臨床栓査. 46. 909-918 (2002)

Noriharu Yosobo、Gen Yamada 等：“肺部炎症的免疫学分析方法”临床研究 46. 909-918 (2002)。

四十坊典晴ほか: "肺クララ細胞10kDa蛋白質(CC10)と免疫病態"Annual Review免疫. 113-119 (2003)

Noriharu Yosobo 等人：“肺 Clara 细胞 10kDa 蛋白 (CC10) 和免疫病理学”年度免疫学评论 113-119 (2003)。