Cytokine-mediated downregulation of coxsackievirus-adenovirus receptor in endothelial cells

Theresa Vincent, Ralf F. Pettersson, Ronald Crystal, Philip L. Leopold

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

Endothelial cells have the ability to change their complement of cell surface proteins in response to inflammatory cytokines. We hypothesized that the expression of the coxsackievirus-adenovirus receptor (CAR), a viral receptor and putative cell-cell adhesion molecule, may be altered during the response of endothelial cells to inflammation. To test this hypothesis, we evaluated CAR protein and mRNA levels in human umbilical vein endothelial cells after they were exposed to tumor necrosis factor alpha, gamma interferon, or a combination of the two cytokines. Flow cytometric and Western blot analyses indicated that cytokine treatment led to a synergistic decrease in CAR protein expression. A Western blot analysis showed that CAR levels decreased to 16% ± 4% or 1% ± 4% of the CAR protein levels in untreated cells with either 24 or 48 h of cytokine treatment, respectively. Quantitative reverse transcription-PCR demonstrated that the combination treatment caused CAR mRNA levels to decrease to 21% ± 12% or 5% ± 3% of the levels in untreated cells after a 24- or 48-h cytokine treatment, respectively. Reduced CAR expression led to a decrease in adenovirus (Ad) binding of 80% ± 3% (compared with untreated endothelial cells), with a subsequent decrease in Ad-mediated gene transfer that was dependent on the dose and duration of cytokine treatment but not on the dose of Ad. A similar decrease in CAR protein level and susceptibility to Ad infection was observed in human microvascular endothelial cells, while CAR expression on normal human bronchial epithelial cells or A549 lung epithelial cells was less affected by cytokine treatments. Taken together, the data demonstrate that inflammatory cytokines decrease CAR mRNA and protein expression with a concomitant decrease in Ad binding, reflecting the impact of cell physiology on the function of CAR and the potential effect of inflammation on the ability of Ad to transfer genes to endothelial cells.

Original languageEnglish
Pages (from-to)8047-8058
Number of pages12
JournalJournal of Virology
Volume78
Issue number15
DOIs
Publication statusPublished - 1 Aug 2004
Externally publishedYes

Fingerprint

Enterovirus
Down-Regulation
Endothelial Cells
Cytokines
Adenoviridae
Proteins
Messenger RNA
adenovirus receptor
Western Blotting
Epithelial Cells
Inflammation
Adenoviridae Infections
Cell Physiological Phenomena
Human Umbilical Vein Endothelial Cells
Cell Adhesion Molecules
Interferon-alpha
Genes
Reverse Transcription
Interferon-gamma
Membrane Proteins

ASJC Scopus subject areas

  • Immunology

Cite this

Cytokine-mediated downregulation of coxsackievirus-adenovirus receptor in endothelial cells. / Vincent, Theresa; Pettersson, Ralf F.; Crystal, Ronald; Leopold, Philip L.

In: Journal of Virology, Vol. 78, No. 15, 01.08.2004, p. 8047-8058.

Research output: Contribution to journalArticle

Vincent, Theresa ; Pettersson, Ralf F. ; Crystal, Ronald ; Leopold, Philip L. / Cytokine-mediated downregulation of coxsackievirus-adenovirus receptor in endothelial cells. In: Journal of Virology. 2004 ; Vol. 78, No. 15. pp. 8047-8058.
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