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型细胞表达基因及其产物在肺疾病病理生理分析、诊断和治疗中的临床应用。本研究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，并将肺凝集素和模式识别受体应用于呼吸道感染的治疗。(1)当检测A549细胞系与IIP患者血清的免疫反应性时，约50%的患者血清与A549细胞呈阳性反应。共聚焦显微镜显示，患者血清识别的抗原定位于细胞的胞质中。患者IgG免疫沉淀120 kDa蛋白。120 kDa蛋白质的蛋白质组学分析鉴定抗原为丙氨酰tRNA合成酶。我们克隆了丙氨酰tRNA合成酶的DNA。当我们用免疫细胞化学的方法 ...更多信息 已显示对患者IgG呈阴性染色并用该酶的cDNA转染的CHO细胞，患者IgG识别与GFP标记的丙氨酰tRNA合成酶共定位的CHO细胞中的抗原。此外，IIP患者的血清显著抑制丙氨酰tRNA合成酶的酶活性。这些结果清楚地证明了IIP患者血清中存在抗丙氨酰tRNA合成酶的自身抗体。(2)放射性肺炎患者血清SP-A、SP-D升高。SP-A和SP-D似乎对能够通过CT检测到但不能通过胸部Xp检测到的小的肺损伤有反应，证明了SP-A和SP-D作为疾病标志物的血清学有用性。此外，我们还建立了肺腺癌患者外周血SP-A、SP-C和CC 10的RT-PCR检测体系。(3)我们已经分析了Toll样受体2的结构-功能关系，该受体已被证明与SP-A和SP-D相互作用。并鉴定了TLR 2的Ser 40-Ile 64区域为葡萄球菌肽聚糖识别和信号转导的功能区域。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