[Detection of HNP1 gene expression by RT-PCR in transfected tracheal epithelial cells].
By using lipofection reagent DOSPER, the plasmid vector carrying the human neutrophil defensin HNP1 gene(pBabe-Neo-HNP1) was transferred into human, rabbit tracheal epithelial cells cultured in serum-free complement medium. Determination of HNP1 mRNA expression in tracheal epithelial cells was performed by reverse transcription-polymerase chain reaction (RT-PCR). The result showed that HNP1-gene expression was detected both in human and rabbit transfected epithelial cells; while in untransfected epithelial cells, no specific signals were detected by RT-PCR. This result and the result of our previous determination by immunocytochemical staining both demonstrated that the epithelial cells could express the antibiotic peptide when transfected with its gene, suggesting that the mucosal antibiotic peptide gene transfer might develop a new antimicrobial approach to prevention and treatment of respiratory infections.
Wu
Q
Q
Research Unit of Infection & Immunity, School of Basic Medical Sciences, WCUMS, Chengdu 610041.
Tang
B
B
Huang
N
N
Wang
B
B
D000818:Animals; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D012333:RNA, Messenger / Q000235:genetics; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014132:Trachea / Q000166:cytology / Q000378:metabolism*; D014162:Transfection; D023084:alpha-Defensins / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics*
D012333:RNA, Messenger; D023084:alpha-Defensins; C406413:human neutrophil peptide 1