Antisense oligonucleotides-induced local blockade of T-bet expression leads to airway inflammation in rats.
OBJECTIVE
To explore whether local blockade of T-box expressed in T cells (T-bet) expression in the lungs could lead to airway inflammation.


METHODS
Twenty-four rats were randomly divided into 4 groups: saline group, ovalbumin (OVA)-sensitized group, nonsense group, and the antisense group. The OVA-sensitized rats were sensitized and challenged with OVA, and the rats in the nonsense and antisense groups were subjected to an aerosol delivery of the nonsense and antisense oligonucleotides (AS-ODN) of T-bet (0.1%, w/v). The levels of interferon-gamma (IFN-gamma), interleukin-4 (IL-4), and IL-5 in the bronchoalveolar lavage fluid (BALF) were detected by ELISA, and the mRNA and the protein expression of T-bet and GATA-3 genes were examined by in situ hybridization and Western blot analysis, respectively.


RESULTS
The expression of T-bet mRNA and protein in the lungs of the rats in the antisense group were inhibited effectively. The lungs of the rats in the antisense and OVA-sensitized groups showed eosinophil and lymphocyte inflammatory infiltration, and eosinophilia located predominantly around the airways. The number of GATA-3 mRNA-positive cells and the level of GATA-3 protein in the lungs of the rats in the antisense and the OVA-sensitized groups significantly increased. The level of IL-4 and IL-5 in the BALF in the antisense and OVA-sensitized groups were elevated, but the level of IFN-gamma decreased markedly.


CONCLUSIONS
Antisense ODN-induced local blockade of T-bet expression leads to airway inflammation with a selective alteration in patterns of cytokine expression and recruitment of eosinophil cells similar to that in the OVA-sensitized animals.
Wang
Gang
G
Department of Integrated Traditional Chinese and Western Medicine, West China Hospital, Sichuan University, Chengdu 610041, China.
Liu
Chun-Tao
CT
Wang
Zeng-Li
ZL
Jiang
Li-Li
LL
Yan
Cun-Liang
CL
Luo
Feng-Min
FM
D000818:Animals; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry; D005260:Female; D050990:GATA3 Transcription Factor / Q000096:biosynthesis* / Q000235:genetics; D007371:Interferon-gamma / Q000378:metabolism; D015847:Interleukin-4 / Q000378:metabolism; D015848:Interleukin-5 / Q000378:metabolism; D008168:Lung / Q000378:metabolism* / Q000473:pathology; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D010047:Ovalbumin / Q000276:immunology; D011014:Pneumonia / Q000378:metabolism* / Q000473:pathology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011897:Random Allocation; D051381:Rats; D017208:Rats, Wistar; D020825:T-Box Domain Proteins / Q000096:biosynthesis* / Q000235:genetics; D000098815:T-bet Transcription Factor
D050990:GATA3 Transcription Factor; D015848:Interleukin-5; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D020825:T-Box Domain Proteins; D000098815:T-bet Transcription Factor; D015847:Interleukin-4; D007371:Interferon-gamma; D010047:Ovalbumin