Induction of endogenous genes following infection of human endothelial cells with an E1- E4+ adenovirus gene transfer vector

Ramachandran Ramalingam, Shahin Rafii, Stefan Worgall, Neil R. Hackett, Ronald Crystal

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Recombinant adenovirus (Ad) gene transfer vectors are effective at transferring exogenous genes to a variety of cells and tissue types both in vitro and in vivo. However, in the process of gene transfer, the Ad vectors induce the expression of target cell genes, some of which may modify the function of the target cell and/or alter the local milieu. To develop a broader understanding of Ad vector-mediated induction of endogenous gene expression, genes induced by first-generation E1- E4+ Ad vectors in primary human umbilical vein endothelial cells were identified by cDNA subtraction cloning. The identified cDNAs included signaling molecules (lymphoid blast crisis [LBC], guanine nucleotide binding-protein et type s [Gα-S], and mitogen kinase [MEK5]), calcium-regulated/cytoskeletal proteins (caipactin p11 and p36 subunits, vinculin, and spinocerebellar ataxia [SCA1]), growth factors (insulin-like growth factor binding protein 4 and transforming growth factor β2), glyceraldehyde-6-phosphate dehydrogenase, an expressed sequence tag, and a novel cDNA showing homology to a LIM domain sequence. Two- to sevenfold induction of the endogenous gene expression was observed at 24 h postinfection, and induction continued up to 72 h, although the timing of gene expression varied among the identified genes. In contrast to that observed in endothelial cells, the Ad vector-mediated induction of gene expression was not found following Ad vector infection of primary human dermal fibroblasts or human alveolar macrophages. Empty Ad capsids did not induce endogenous gene expression in endothelial cells. Interestingly, additional deletion of the E4 gene obviated the upregulation of genes in endothelial cells by the El- E3- Ad vector, suggesting that genes carried by the E4 region play a central role in modifying target cell gene expression. These findings are consistent with the notion that efficient transfer of exogenous genes to endothelial cells by first-generation Ad vectors comes with the price that these vectors also induce the expression of a variety of cellular genes.

Original languageEnglish
Pages (from-to)10183-10190
Number of pages8
JournalJournal of Virology
Volume73
Issue number12
Publication statusPublished - 1 Dec 1999
Externally publishedYes

Fingerprint

Adenoviridae
Endothelial Cells
Infection
Genes
Gene Expression
Spinocerebellar Ataxias
Complementary DNA
Insulin-Like Growth Factor Binding Protein 4
Glyceraldehyde
Vinculin
Blast Crisis
Adenoviridae Infections
Cytoskeletal Proteins
Guanine Nucleotides
Capsid
Expressed Sequence Tags
Human Umbilical Vein Endothelial Cells
Gene Deletion
Alveolar Macrophages
Transforming Growth Factors

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Induction of endogenous genes following infection of human endothelial cells with an E1- E4+ adenovirus gene transfer vector. / Ramalingam, Ramachandran; Rafii, Shahin; Worgall, Stefan; Hackett, Neil R.; Crystal, Ronald.

In: Journal of Virology, Vol. 73, No. 12, 01.12.1999, p. 10183-10190.

Research output: Contribution to journalArticle

Ramalingam, Ramachandran ; Rafii, Shahin ; Worgall, Stefan ; Hackett, Neil R. ; Crystal, Ronald. / Induction of endogenous genes following infection of human endothelial cells with an E1- E4+ adenovirus gene transfer vector. In: Journal of Virology. 1999 ; Vol. 73, No. 12. pp. 10183-10190.
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