Proteomic analysis of peripheral T-lymphocytes in patients with asthma.
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 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 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.
Jeong
Hye Cheol
HC
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.
Lee
Sang Yeub
SY
Lee
Eun Joo
EJ
Jung
Ki Hwan
KH
Kang
Eun Hae
EH
Lee
Sung Yong
SY
Kim
Je Hyeong
JH
Park
Eun Kyung
EK
Lee
Sang Hoon
SH
Uhm
Chang Sub
CS
Cho
Yunjung
Y
Shin
Chol
C
Shim
Jae Jeong
JJ
Kim
Han Kyeom
HK
In
Kwang Ho
KH
Kang
Kyung Ho
KH
Yoo
Se Hwa
SH
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000235:genetics / Q000378:metabolism; D000328:Adult; D001249:Asthma / Q000097:blood* / Q000276:immunology; D015415:Biomarkers / Q000378:metabolism; D017252:CD3 Complex / Q000276:immunology; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D015870:Gene Expression; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011379:Prognosis; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D012720:Severity of Illness Index; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism*; D013879:Thioredoxins / Q000235:genetics / Q000378:metabolism
D015415:Biomarkers; D017252:CD3 Complex; D008565:Membrane Proteins; D011506:Proteins; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; C516110:TRX2 protein, S cerevisiae; D013879:Thioredoxins; C483009:GSTM3 protein, human; D005982:Glutathione Transferase; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4; C517361:PDE4C protein, human