[In vivo transfection of cis element 'decoy' against nuclear factor-kappaB binding site modulates function of T lymphocytes in asthma].
OBJECTIVE
To investigate the effect of nuclear factor-kappaB (NF-kappaB) decoy oligodeoxynucleotide (ODN) on proliferation, apoptosis and cytokine synthesis of T lymphocytes from asthmatic patients.


METHODS
T lymphocytes were divided into four groups, a normal control group (A group), an asthma control group (B group), a NF-kappaB cis decoy ODN group (B(1) group) and a scrambled ODN group (B(2) group). B(1) and B(2) groups were transfected by cationic lipofectamine to the latter two groups respectively. The proliferation of T lymphocytes was measured by MTT and the apoptosis of them was measured by flow cytometry. The expression levels of interleukin 5 (IL-5) mRNA and protein were detected with cell hybridization in situ and enzyme-linked immunosorbent assay (ELISA). The expression levels of inducible nitric oxide synthase (iNOS) were measured by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.


RESULTS
The difference of proliferation rate between B(1) group (0.220 +/- 0.020) and B group (0.340 +/- 0.030) was significant (P < 0.05). The difference of apoptosis rate between B(1) group (10.8 +/- 1.3) and B group (8.1 +/- 1.2) was also significant (P < 0.05). The expression levels of IL-5 mRNA and protein in B(1) group (21 +/- 4, 24 +/- 4) were significantly different from those in B group (33 +/- 4, 54 +/- 10, P < 0.05). The differences of iNOS mRNA and protein levels between B(1) group (0.33 +/- 0.05, 782 +/- 117) and B group (0.75 +/- 0.13, 1185 +/- 230) were significant (P < 0.05). However, these indices in B(2) group showed no difference to those in B group (P > 0.05).


CONCLUSIONS
NF-kappaB decoy ODN can reduce the abnormally increased proliferation of T lymphocytes in asthma and increase the abnormally decreased apoptosis of these cells, while decrease the abnormally increased levels of cytokine and enzyme in asthmatic T lymphocytes. These may be the mechanisms underlying its potential therapeutic effects in asthma.
Department of Respiratory Medicine, Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Ning
N
Zhang
Yong-jian
YJ
Xu
Zhen-xiang
ZX
Zhang
Wei-ning
WN
Xiong
D017209:Apoptosis / Q000235:genetics / Q000502:physiology; D001249:Asthma / Q000503:physiopathology*; D001665:Binding Sites / Q000235:genetics; D015153:Blotting, Western; D002455:Cell Division / Q000235:genetics / Q000502:physiology; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D006801:Humans; D015848:Interleukin-5 / Q000032:analysis; D016328:NF-kappa B / Q000235:genetics* / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000235:genetics; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000378:metabolism*; D014162:Transfection
D015848:Interleukin-5; D016328:NF-kappa B; D009838:Oligodeoxyribonucleotides; D019001:Nitric Oxide Synthase; D016650:Fluorescein-5-isothiocyanate