哮喘患者外周T淋巴细胞蛋白质组分析
2008/02/07
哮喘是以可逆性气流受阻为特征的慢性气道炎症性疾病,其中T淋巴细胞在炎症中起重要作用。蛋白质组分析技术是目前发展迅速的一项研究手段，被广泛应用于基因研究领域，是基因研究的重要组成部分。韩国学者Hye Cheol Jeong等通过标准蛋白质组技术（包括2D-PAGE,MALDI-TOF-MS和数据库搜索）对哮喘患者T淋巴细胞进行了研究。研究选取了6名从未使用过激素的哮喘患者和6名健康人作为对照，通过2D－PAGE电泳和银染色点分析比较，再以MALDI-TOF-MS和数据库搜索进行确认，并选取某些确认的蛋白质进行mRNA实时PCR检测。
结果显示，与正常人相比，共有13个位点的蛋白质表达较正常人增高，12个位点的蛋白质表达较正常人降低。对于可确认的蛋白质进行mRNA的进一步分析显示磷酸二酯酶4C表达增高，谷胱甘肽S转移酶M3的表达降低。
因此，蛋白质组研究有助于发现哮喘患者外周血T淋巴细胞的不同表达位点，并可能将这些不同表达的蛋白质作为今后哮喘研究的重要的生物活性指标和治疗评价指标。
（汤葳 上海交通大学附属瑞金医院呼吸科 200025 摘译）
(Chest. 2007; 132:489-496)
Proteomic Analysis of Peripheral T-Lymphocytes in
Patients With Asthma*Hye Cheol Jeong, MD, PhD; Sang Yeub Lee, MD, PhD; Eun Joo Lee, MD;
Ki Hwan Jung, MD; Eun Hae Kang, MD; Sung Yong Lee, MD, PhD; Je
Hyeong Kim, MD, PhD; Eun Kyung Park, PhD; Sang Hoon Lee; Chang Sub
Uhm,MD, PhD; Yunjung Cho, MD, PhD; Chol Shin, MD, PhD, FCCP; Jae
Jeong Shim, MD, PhD; Han Kyeom Kim, MD, PhD; Kwang Ho In, MD, PhD;
Kyung Ho Kang, MD, PhD, FCCP and Se Hwa Yoo, MD, PhD * From the Division of Respiratory and Critical Care Medicine (Dr.
Jeong), Department of Internal Medicine, College of Medicine, Pochon
CHA University,Seongnam; Division of Respiratory and Critical Care
Medicine (Drs. Sang Yeub Lee, E. J. Lee, Kang, In, and Yoo),Department
of Internal Medicine, Anam Hospital, Korea University, Seoul; Division
of Respiratory and Critical Care Medicine (Drs. Sung Yong Lee, Shim,
and Kang), Department of Internal Medicine, Guro Hospital, Korea
University, Seoul;Department of Internal Medicine (Drs. Jung, J. H. Kim,
and Shin),Ansan Hospital, Korea University, Ansan; Department of Anatomy
(Drs. Park, Uhm, and Mr. S. H. Lee), College of Medicine, Korea University,
Seoul; Department of Laboratory Medicine (Dr. Cho), College of Medicine,
Korea University, Seoul; and Department of Pathology (Dr. H. K. Kim), College of Medicine, Korea University, Seoul, Korea.
Correspondence to: Sang Yeub Lee, MD, PhD, Division of Respiratory and
Critical Care Medicine; Department of Internal Medicine, College of
Critical Care Medicine; Department of Internal Medicine, College of
Medicine,Korea University, 126–1, 5ga Anam Dong, Seongbuk gu, Seoul,
136–705, South Korea; e-mail: pulsy@korea.ac.kr Abstract
Background: 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.
Methods: The proteins of CD3+ T-lymphocytes were isolated from whole
Methods: 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). Results: 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.
Conclusions: Proteomic examination of the peripheral T-lymphocytes
Conclusions: 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. Key Words: asthma • proteomics • T-lymphocytes
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IL-4变体对哮喘患者变应原导致的迟发哮喘反应的作用
