Selective targeting of human cells by a chimeric adenovirus vector containing a modified fiber protein.
The adenovirus fiber protein is responsible for attachment of the virion to unidentified cell surface receptors. There are at least two distinct adenovirus fiber receptors which interact with the group B (Ad3) and group C (Ad5) adenoviruses. We have previously shown by using expressed adenovirus fiber proteins that it is possible to change the specificity of the fiber protein by exchanging the head domain with another serotype which recognizes a different receptor (S. C. Stevenson et al., J. Virol. 69:2850-2857, 1995). A chimeric fiber cDNA containing the Ad3 fiber head domain fused to the Ad5 fiber tail and shaft was incorporated into the genome of an adenovirus vector with E1 and E3 deleted encoding beta-galactosidase to generate Av9LacZ4, an adenovirus particle which contains a chimeric fiber protein. Western blot analysis of the chimeric fiber vector confirmed expression of the chimeric fiber protein and its association with the adenovirus capsid. Transduction experiments with fiber protein competitors demonstrated the altered receptor tropism of the chimeric fiber vector compared to that of the parental Av1LacZ4 vector. Transduction of a panel of human cell lines with the chimeric and parental vectors provided evidence for a different cellular distribution of the Ad5 and Ad3 receptors. Three cell lines (THP-1, MRC-5, and FaDu) were more efficiently transduced by the vector containing the Ad3 fiber head than by the Ad5 fiber vector. In contrast, human coronary artery endothelial cells were transduced more readily with the vector containing the Ad5 fiber than with the chimeric fiber vector. HeLa and human umbilical vein endothelial cells were transduced at equivalent levels compared with human diploid fibroblasts, which were refractory to transduction with both vectors. These results provide evidence for the differential expression of the Ad5 and Ad3 receptors on human cell lines derived from clinically relevant target tissues. Furthermore, we show that exchange of the fiber head domain is a viable approach to the production of adenovirus vectors with cell-type-selective transduction properties. It may be possible to extend this approach to the use of ligands for a range of different cellular receptors in order to target gene transfer to specific cell types at the level of transduction.
Stevenson
S C
SC
Department of Molecular and Cell Biology, Genetic Therapy, Inc., Gaithersburg, Maryland 20878, USA.
Rollence
M
M
Marshall-Neff
J
J
McClelland
A
A
D000256:Adenoviridae / Q000235:genetics*; D002213:Capsid / Q000235:genetics*; D036022:Capsid Proteins; D002460:Cell Line; D005814:Genes, Viral; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014161:Transduction, Genetic; D015678:Viral Structural Proteins / Q000235:genetics
D036022:Capsid Proteins; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins; D015678:Viral Structural Proteins; C018315:hexon capsid protein, Adenovirus