Jeong Hye Cheol, Lee Sang Yeub, Lee Eun Joo, Jung Ki Hwan, Kang Eun Hae, Lee Sung Yong, Kim Je Hyeong, Park Eun Kyung, Lee Sang Hoon, Uhm Chang Sub, Cho Yunjung, Shin Chol, Shim Jae Jeong, Kim Han Kyeom, In Kwang Ho, Kang Kyung Ho, Yoo Se Hwa
Division of Respiratory and Critical Care Medicine; Department of Internal Medicine, College of Medicine, Korea University, 126-1, 5ga Anam Dong, Seongbuk gu, Seoul, 136-705, South Korea.
Chest. 2007 Aug;132(2):489-96. doi: 10.1378/chest.06-2980. Epub 2007 Jun 5.
Asthma is chronic airway inflammation that occurs together with reversible airway obstruction. T-lymphocytes play an important role in the pathogenesis of asthma. Proteomic technology has rapidly developed in the postgenomic era, and it is now widely accepted as a complementary technology to genetic profiling. We investigated the changes of proteins in T-lymphocytes of asthma patients by using standard proteome technology: two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), and a database search.
The proteins of CD3+ T-lymphocytes were isolated from whole blood of six steroid-naive asthmatic patients and of six healthy volunteers. 2D-PAGE was performed and the silver-stained protein spots were comparatively analyzed between the asthma and control groups using an image analyzer. Some differentially expressed spots were identified by MALDI-TOF-MS and database search. The messenger RNA expressions of some identified proteins were examined by real-time polymerase chain reaction (RT-PCR).
Thirteen protein spots in the T-lymphocytes of the asthmatic patients were increased and 12 spots were decreased compared to those of the normal subjects. Among the identified proteins, the increased expression of the messenger RNA of phosphodiesterase 4C and thioredoxin-2 and the decreased expression of the messenger RNA of glutathione S-transferase M3 were confirmed by RT-PCR in the asthmatic patients.
Proteomic examination of the peripheral T-lymphocytes revealed some differentially expressed proteins in the asthmatic patients. The possibility of using the differentially expressed proteins as important biomarkers and therapeutic targets in asthma patients warrants further studies.
哮喘是一种伴有可逆性气道阻塞的慢性气道炎症。T淋巴细胞在哮喘发病机制中起重要作用。蛋白质组学技术在后基因组时代迅速发展，现已被广泛接受为基因谱分析的补充技术。我们采用标准蛋白质组技术，即二维聚丙烯酰胺凝胶电泳（2D-PAGE）、基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）和数据库检索，研究哮喘患者T淋巴细胞中蛋白质的变化。
从6例未使用类固醇的哮喘患者和6例健康志愿者的全血中分离CD3+T淋巴细胞的蛋白质。进行2D-PAGE，并使用图像分析仪对哮喘组和对照组之间的银染蛋白斑点进行比较分析。通过MALDI-TOF-MS和数据库检索鉴定一些差异表达的斑点。通过实时聚合酶链反应（RT-PCR）检测一些鉴定出的蛋白质的信使核糖核酸表达。
与正常受试者相比，哮喘患者T淋巴细胞中有13个蛋白斑点增加，12个斑点减少。在鉴定出的蛋白质中，哮喘患者经RT-PCR证实磷酸二酯酶4C和硫氧还蛋白-2的信使核糖核酸表达增加，谷胱甘肽S-转移酶M3的信使核糖核酸表达减少。
外周血T淋巴细胞的蛋白质组学检查揭示了哮喘患者中一些差异表达的蛋白质。将差异表达的蛋白质用作哮喘患者重要生物标志物和治疗靶点的可能性值得进一步研究。
