Adenovirus-mediated persistent cystic fibrosis transmembrane conductance regulator expression in mouse airway epithelium.
Replication-defective adenovirus (Ad) vectors have been used for gene transfer to the respiratory epithelium of experimental animals and individuals with cystic fibrosis. Studies from several laboratories have suggested that administration of first-generation Ad vectors results only in transient gene expression in the lung, due at least in part to destruction of vector-transduced cells by host cellular immune responses directed against viral proteins and/or immunogenic transgene products. We have constructed new Ad2-based, E1-deleted vectors encoding a weakly immunogenic transgene, the human cystic fibrosis transmembrane conductance regulator (hCFTR) under the control of the cytomegalovirus enhancer-promoter. These vectors contain wild-type E2 and E4 regions. These new Ad/CFTR vectors were instilled into the lungs of immunocompetent C57BL/6, BALB/c, and C3H mice. In vitro cytotoxic T lymphocyte (CTL) analysis indicated the presence of Ad-specific CTLs in treated mice. However, we were not able to demonstrate a CTL response specific for hCFTR. Reverse transcriptase PCR analysis demonstrated that hCFTR mRNA expression continued in all three strains of mice for at least 70 days, the last time point analyzed. The E3 region did not play a significant role in persistence of the Ad/CFTR vectors in the mouse lung. Functional hCFTR expression was also observed in the nasal epithelia of CF mutant mice. These results suggest that long-term expression of hCFTR is possible in the airway epithelia of immunocompetent mice without radical modification of Ad vector and in spite of the presence of CTLs.
Genzyme Corporation, Framingham, Massachusetts 01701, USA. ascaria@genzyme.com
A
A
Scaria
J A
JA
St George
C
C
Jiang
J M
JM
Kaplan
S C
SC
Wadsworth
R J
RJ
Gregory
D017503:Adenovirus E3 Proteins / Q000235:genetics; D000260:Adenoviruses, Human / Q000276:immunology; D000818:Animals; D003001:Cloning, Molecular; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D004848:Epithelium / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000276:immunology; D006801:Humans; D007121:Immunocompetence; D008168:Lung / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
D017503:Adenovirus E3 Proteins; C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator