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

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。我们鉴定了特发性间质性肺炎(IIP)患者血清中存在的自身抗体抗原，确立了血清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)A549细胞株与IIP患者血清免疫反应时，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胞浆。患者免疫沉淀检测到120 kDa蛋白。对120 kDa蛋白进行蛋白质组学分析，确定该抗原为丙氨酰tRNA合成酶。我们已经克隆了丙氨酰tRNA合成酶的DNA。当我们对…进行免疫细胞化学分析时更多的CHO细胞已被证明是阴性的患者免疫球蛋白，并被CDNA为该酶，患者免疫球蛋白识别的CHO细胞与GFP标记的丙氨酰tRNA合成酶共定位的抗原。此外，IIP患者血清对丙氨酰tRNA合成酶活性有明显的抑制作用。结果表明，IIP患者血清中存在抗丙氨酰-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对CT能发现但胸部XP不能发现的微小肺损伤有反应，证明了SP-A和SP-D作为疾病标志物的血清学价值。此外，我们还建立了肺腺癌患者外周血中SP-A、SP-C和CC10的RT-PCR检测体系。(3)分析了与SP-A和SP-D相互作用的Toll样受体2的结构与功能关系。确定TLR2区Ser40-Ile64为葡萄球菌肽聚糖识别和信号转导的功能区。较少

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