Adenoviral interleukin-12 gene transduction into human bronchial epithelial cells: up-regulation of pro-inflammatory cytokines and its prevention by corticosteroids.
BACKGROUND
One of the potential effects of IL-12 is to restore Th1/Th2 balance. Therefore, we investigated the possibility of developing a system for local delivery of IL-12 into the airways by examining protein expression in a human bronchial epithelial cell line (BEAS-2B) after adenoviral IL-12 gene transduction. The effects of dexamethasone on the gene-modified cells were also examined.


METHODS
Adenoviral vectors AxCAegfp and Ax1CIhp40ip35 were used to transduce enhanced green fluorescence protein and IL-12 genes, respectively, into BEAS-2B cells. Wild-type and IL-12 gene-transduced BEAS-2B cells were then incubated with or without dexamethasone, and concentrations of IL-12, IFN-gamma, IL-6, IL-8, granulocyte macrophage-colony stimulating factor and chemokines (TARC and RANTES) in the supernatant were measured by ELISA. IL-12 receptor expression was analysed by flow cytometry and RT-PCR.


RESULTS
The efficiency of transgene expression in BEAS-2B cells at a multiplicity of infection of 30 was approximately 80%. Gene-modified BEAS-2B cells produced biologically active IL-12, regardless of dexamethasone treatment. While IL-12 gene transduction led to increased production of IL-6 and IL-8 by BEAS-2B cells, expressions of these proteins were suppressed by dexamethasone. Addition of exogenous IL-12 failed to augment BEAS-2B cell IL-6 and IL-8 production, and IL-12 receptor expression by BEAS-2B cells was not detected.


CONCLUSIONS
Our findings suggest that adenoviral IL-12 gene transduction may be effective in inducing IL-12 expression in the airways, and could be a potential approach in the management of bronchial asthma.
Ogawa
H
H
Department of Internal Medicine and Molecular Therapeutics, University of Tokushima, School of Medicine, Tokushima, Japan.
Nishimura
N
N
Nishioka
Y
Y
Azuma
M
M
Yanagawa
H
H
Sone
S
S
D000256:Adenoviridae; D001980:Bronchi; D002460:Cell Line; D003907:Dexamethasone / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005822:Genetic Vectors; D005938:Glucocorticoids / Q000494:pharmacology*; D006801:Humans; D018664:Interleukin-12 / Q000235:genetics*; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D020545:Respiratory Mucosa; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014161:Transduction, Genetic; D015854:Up-Regulation
D005938:Glucocorticoids; D015850:Interleukin-6; D016209:Interleukin-8; D018664:Interleukin-12; D012313:RNA; D003907:Dexamethasone