Efficient adenovirus-mediated gene transfer to gastric tissue by oral administration.
BACKGROUND
Recombinant adenoviruses (rAd) are well-characterized viral vectors and have been studied in many human diseases. However, there are no detailed methods for transferring genes to the stomach using rAd.


METHODS
Gastric epithelial cells were infected with rAd encoding green fluorescence protein (AdGFP) for different times, or with AdGFP that had been incubated in artificial gastric juice at different pH values for 1 h. Gene expression was detected by fluorescence microscope and flow cytometry. Mice were infected via oral administration with rAd encoding red fluorescence protein and beta-galactosidase (AdRFP-lacZ) or rAd encoding mouse interleukin-17 (AdmIL-17), and tissues were collected at the indicated times after infection. LacZ expression in different tissues was detected by X-gal staining and IL-17 expression in the stomach was assessed by the real-time polymerase chain reaction and an enzyme-linked immunosorbent assay. Inflammation in the stomach was also assessed.


RESULTS
rAd could infect the gastric epithelial cells and tolerate pH 5 for 1 h in vitro. Adenovirus-mediated genes were specifically expressed in the gastrointestinal tract and transgene expression persisted in gastric tissue for up to 7 days after oral administration of AdRFP-lacZ. Oral administration of AdmIL-17 induced mIL-17 expression in gastric tissue at the mRNA and protein levels and protein level peaked on day 5 post-infection. IL-6, a target protein of IL-17, and gastric inflammation also increased in AdmIL-17-infected mice.


CONCLUSIONS
The present study has established a detailed method for transferring adenovirus-mediated gene to the stomach, which may provide a valuable approach for gene therapy or the study of the basic biology of gastric diseases.
Liu
Xiao-Fei
XF
Department of Clinical Microbiology and Immunology, Third Military Medical University, Chongqing, China.
Shi
Yun
Y
Zhang
Jin-Yu
JY
Zhuang
Yuan
Y
Jia
Ke-Ran
KR
Mao
Xu-Hu
XH
Guo
Ying
Y
Liu
Tao
T
Liu
Zhen
Z
Wu
Chao
C
Zhang
Wei-Jun
WJ
Zhou
Wei-Ying
WY
Guo
Gang
G
Zou
Quan-Ming
QM
D000256:Adenoviridae / Q000235:genetics*; D000284:Administration, Oral; D000818:Animals; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D041981:Gastrointestinal Tract; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D020381:Interleukin-17 / Q000235:genetics; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D007763:Lac Operon / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013274:Stomach Neoplasms / Q000235:genetics* / Q000821:virology; D019076:Transgenes / Q000502:physiology; D001616:beta-Galactosidase / Q000235:genetics; D000097573:Red Fluorescent Protein
D020381:Interleukin-17; D015850:Interleukin-6; D008164:Luminescent Proteins; D012333:RNA, Messenger; D001616:beta-Galactosidase; D000097573:Red Fluorescent Protein