Clinical application for pathophysiological analysis, diagnosis and treatment of lung diseases by genes and their products expressed in alveolar type II cells.

肺泡II型细胞表达的基因及其产物在肺部疾病病理生理学分析、诊断和治疗中的临床应用。

• 批准号: 12557057
• 负责人: KUROKI Yoshio
• 金额: $ 8.51万
• 依托单位: Sapporo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557057/
• 关键词: alveolar type II cells; idiopathic interstitial pneumonia; SP-A; SP-D; radiation pneumonitis; autoantibody; Toll-like receptor; lung adenocarcinoma; A549細胞; 肺サーファクタント蛋白質; SP-C; 放射線肺臓炎; 膠原病合併間質性肺炎; 生体防御

The purpose of this study was to pursue clinical application for pathophysiological analysis, diagnosis and treatment of lung diseases by genes and their products expressed in alveolar type II cells. We have identified the antigen to autoantibody existing in sera from patients with idiopathic interstitial pneumonia (IIP), established clinical significance of serum SP-A and SP-D as disease markers and applied lung collectins and pattern recognition receptors for treatment of respiratory infections.(1) When A549 cell line was examined for immunoreactivity with sera from patients with IIP, approximately 50 % of patient sera reacted positively with A549 cells. Confocal microscopy revealed that the antigen recognized by patient sera localized in the cyoplasm of the cells. Patient IgG immunoprecipitaed 120 kDa protein. Proteome analysis of the 120 kDa protein identified the antigen as alanyl tRNA synthetase. We have cloned DNA for alanyl tRNA synthetase. When we analyzed immunocytochemically … More CHO cells which had been shown to be negative staining for patient IgG and were transfected with CDNA for this enzyme, patient IgG recognized the antigen in CHO cells that colocalized with GFP-labeled alanyl tRNA synthetase. In addition, sera from IIP patients significantly inhibited the enzyme activities of alanyl tRNA synthetase. These results clearly demonstrate that there exists antoantibody to alanyl tRNA synthetase in sera from IIP patients.(2) Serum SP-A and SP-D increased in patients with radiation pneumonitis. SP-A and SP-D appeared to respond to the small lung damages which was able to detect by CT but not by chest Xp, demonstrating the serological usefulness of SP-A and SP-D as disease markers. In addition, we have establised the detection system of micrometastasis in peripheral blood in patients with lung adenocarcinoma using RT-PCR for SP-A, SP-C and CC10.(3) We have analyzed the structure-function relationship of Toll-like receptor 2 that has been shown to interact with SP-A and SP-D. and identified the TLR2 region Ser40-Ile64 as the functional region in recognition and signal transduction of Staphylococcal peptidoglycan. Less

本研究旨在探讨肺泡II型细胞表达的基因及其产物在肺部疾病病理生理分析、诊断和治疗中的临床应用。我们确定了特发性间质性肺炎（IIP）患者血清中存在的抗原对自身抗体，确定了血清SP-A和SP-D作为疾病标志物的临床意义，并应用肺集合素和模式识别受体治疗呼吸道感染。(1)当检测A549细胞系与IIP患者血清的免疫反应性时，大约50%的患者血清与A549细胞反应阳性。共聚焦显微镜显示，患者血清识别的抗原定位于细胞的细胞质中。患者IgG免疫沉淀120 kDa蛋白。120 kDa蛋白的蛋白质组学分析鉴定抗原为丙烯酰tRNA合成酶。我们克隆了丙烯酰tRNA合成酶的DNA。当我们分析免疫细胞化学时，更多的CHO细胞被证明对IgG呈阴性染色，并转染了该酶的CDNA，患者IgG识别了与gfp标记的丙烯酰tRNA合成酶共定位的CHO细胞中的抗原。此外，IIP患者血清显著抑制丙烯酰tRNA合成酶的酶活性。这些结果清楚地表明，IIP患者血清中存在丙烯酰tRNA合成酶抗体。(2)放射性肺炎患者血清SP-A、SP-D升高。SP-A和SP-D似乎对CT检测到的小肺损伤有反应，但胸部Xp检测不到，这表明SP-A和SP-D作为疾病标志物的血清学有用。此外，我们建立了肺腺癌患者外周血微转移检测系统，采用RT-PCR检测SP-A、SP-C和CC10。(3)我们分析了与SP-A和SP-D相互作用的toll样受体2的结构-功能关系。并鉴定出TLR2区域Ser40-Ile64是葡萄球菌肽聚糖识别和信号转导的功能区。少

项目成果

Mitsuzawa H.: "Extracellular Toll-like receptor 2 region containing Ser40-Ile64 but not Cys30-Ser39 is critical for the recognition of Staphylococcus aur bus peptidoglycan"J. Biol. Chem.. 276. 6830-6837 (2002)

Mitsuzawa H.：“含有 Ser40-Ile64 但不含 Cys30-Ser39 的细胞外 Toll 样受体 2 区域对于识别金色葡萄球菌肽聚糖至关重要”

Takahashi H: "Serum levels of surfactant proteins A and D are useful markers for interstitial lung disease in patients with progressive systemic sclerosis."Am J Respir Crit Care Med. 162. 258-263 (2000)

Takahashi H：“表面活性蛋白 A 和 D 的血清水平是进行性系统性硬化症患者间质性肺疾病的有用标志物。”Am J Respir Crit Care Med。

高橋亨: "GM-CSFノックアウトマウス"分子呼吸器病. (印刷中). (2001)

Toru Takahashi：“GM-CSF 敲除小鼠”分子呼吸道疾病（印刷中）。

Cheng G: "Increased levels of surfactant protein A and D in bronchoalveolar lavage fluids in patients with bronchial asthma"Eur Respir J. 16. 831-835 (2000)

Cheng G：“支气管哮喘患者支气管肺泡灌洗液中表面活性蛋白 A 和 D 水平升高”Eur Respir J. 16. 831-835 (2000)

Takahashi H: "Serum levels of surfactant protein A and D are useful markers for interstitial lung disease in natients with progressive systemic sclerosis"Am J Respir Crit care Med. 62. 258-263 (2000)

Takahashi H：“表面活性蛋白 A 和 D 的血清水平是进行性系统性硬化症患者间质性肺疾病的有用标志物”Am J Respir Crit care Med。