Transcriptional activation of the interleukin-8 gene by respiratory syncytial virus infection in alveolar epithelial cells: Nuclear translocation of the RelA transcription factor as a mechanism producing airway mucosal inflammation

Roberto Garofalo, Mona Sabry, Mohammad Jamaluddin, Robert K Yu, Antonella Casola, Pearay L. Ogra, Allan R. Brasier

Research output: Contribution to journalArticle

176 Citations (Scopus)

Abstract

The most common cause of epidemic pediatric respiratory disease, respiratory syncytial virus (RSV), stimulates interleukin-8 (IL-8) synthesis upon infecting airway epithelium, an event necessary for the development of mucosal inflammation. We investigated the mechanism for enhanced IL-8 production in human A549 type II pulmonary epithelial cells. Infection with sucrose-purified RSV (pRSV) produced a time-dependent increase in the transcriptional initiation rate of the IL-8 gene. Transient transfection of the human IL-8 promoter mutated in the binding site fur nuclear factor-κB (NF-κB) demonstrated that this sequence was essential for pRSV-activated transcription. Gel mobility shift assays demonstrated pRSV induction of sequence-specific binding complexes; these complexes were supershifted only by antibodies directed to the potent NF-κB transactivating subunit RelA. Both Western immunoblot and indirect immunofluorescence assays showed that cytoplasmic RelA in uninfected cells became localized to the nucleus after pRSV infection. RelA activation requires replicating RSV, because neither conditioned medium nor UV-inactivated pRSV was able to stimulate its translocation. We conclude that RelA undergoes changes in subcellular distribution in airway epithelial cells upon pRSV infection. The ability of replicating RSV to activate RelA translocation may play an important rule in activating IL-8 and other inflammatory gene products necessary for airway mucosal inflammation seen in RSV disease.

Original languageEnglish (US)
Pages (from-to)8773-8781
Number of pages9
JournalJournal of Virology
Volume70
Issue number12
StatePublished - Dec 1 1996

Fingerprint

Transcription Factor RelA
Alveolar Epithelial Cells
Respiratory Syncytial Virus Infections
Respiratory Syncytial Viruses
interleukin-8
transcriptional activation
Interleukin-8
Transcriptional Activation
epithelial cells
transcription factors
inflammation
Inflammation
viruses
infection
Genes
genes
Infection
Epithelial Cells
Electrophoretic Mobility Shift Assay
assays

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Transcriptional activation of the interleukin-8 gene by respiratory syncytial virus infection in alveolar epithelial cells : Nuclear translocation of the RelA transcription factor as a mechanism producing airway mucosal inflammation. / Garofalo, Roberto; Sabry, Mona; Jamaluddin, Mohammad; Yu, Robert K; Casola, Antonella; Ogra, Pearay L.; Brasier, Allan R.

In: Journal of Virology, Vol. 70, No. 12, 01.12.1996, p. 8773-8781.

Research output: Contribution to journalArticle

@article{a05cee4d5b804c97bdb95145f30b337c,
title = "Transcriptional activation of the interleukin-8 gene by respiratory syncytial virus infection in alveolar epithelial cells: Nuclear translocation of the RelA transcription factor as a mechanism producing airway mucosal inflammation",
abstract = "The most common cause of epidemic pediatric respiratory disease, respiratory syncytial virus (RSV), stimulates interleukin-8 (IL-8) synthesis upon infecting airway epithelium, an event necessary for the development of mucosal inflammation. We investigated the mechanism for enhanced IL-8 production in human A549 type II pulmonary epithelial cells. Infection with sucrose-purified RSV (pRSV) produced a time-dependent increase in the transcriptional initiation rate of the IL-8 gene. Transient transfection of the human IL-8 promoter mutated in the binding site fur nuclear factor-κB (NF-κB) demonstrated that this sequence was essential for pRSV-activated transcription. Gel mobility shift assays demonstrated pRSV induction of sequence-specific binding complexes; these complexes were supershifted only by antibodies directed to the potent NF-κB transactivating subunit RelA. Both Western immunoblot and indirect immunofluorescence assays showed that cytoplasmic RelA in uninfected cells became localized to the nucleus after pRSV infection. RelA activation requires replicating RSV, because neither conditioned medium nor UV-inactivated pRSV was able to stimulate its translocation. We conclude that RelA undergoes changes in subcellular distribution in airway epithelial cells upon pRSV infection. The ability of replicating RSV to activate RelA translocation may play an important rule in activating IL-8 and other inflammatory gene products necessary for airway mucosal inflammation seen in RSV disease.",
author = "Roberto Garofalo and Mona Sabry and Mohammad Jamaluddin and Yu, {Robert K} and Antonella Casola and Ogra, {Pearay L.} and Brasier, {Allan R.}",
year = "1996",
month = "12",
day = "1",
language = "English (US)",
volume = "70",
pages = "8773--8781",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "12",

}

TY - JOUR

T1 - Transcriptional activation of the interleukin-8 gene by respiratory syncytial virus infection in alveolar epithelial cells

T2 - Nuclear translocation of the RelA transcription factor as a mechanism producing airway mucosal inflammation

AU - Garofalo, Roberto

AU - Sabry, Mona

AU - Jamaluddin, Mohammad

AU - Yu, Robert K

AU - Casola, Antonella

AU - Ogra, Pearay L.

AU - Brasier, Allan R.

PY - 1996/12/1

Y1 - 1996/12/1

N2 - The most common cause of epidemic pediatric respiratory disease, respiratory syncytial virus (RSV), stimulates interleukin-8 (IL-8) synthesis upon infecting airway epithelium, an event necessary for the development of mucosal inflammation. We investigated the mechanism for enhanced IL-8 production in human A549 type II pulmonary epithelial cells. Infection with sucrose-purified RSV (pRSV) produced a time-dependent increase in the transcriptional initiation rate of the IL-8 gene. Transient transfection of the human IL-8 promoter mutated in the binding site fur nuclear factor-κB (NF-κB) demonstrated that this sequence was essential for pRSV-activated transcription. Gel mobility shift assays demonstrated pRSV induction of sequence-specific binding complexes; these complexes were supershifted only by antibodies directed to the potent NF-κB transactivating subunit RelA. Both Western immunoblot and indirect immunofluorescence assays showed that cytoplasmic RelA in uninfected cells became localized to the nucleus after pRSV infection. RelA activation requires replicating RSV, because neither conditioned medium nor UV-inactivated pRSV was able to stimulate its translocation. We conclude that RelA undergoes changes in subcellular distribution in airway epithelial cells upon pRSV infection. The ability of replicating RSV to activate RelA translocation may play an important rule in activating IL-8 and other inflammatory gene products necessary for airway mucosal inflammation seen in RSV disease.

AB - The most common cause of epidemic pediatric respiratory disease, respiratory syncytial virus (RSV), stimulates interleukin-8 (IL-8) synthesis upon infecting airway epithelium, an event necessary for the development of mucosal inflammation. We investigated the mechanism for enhanced IL-8 production in human A549 type II pulmonary epithelial cells. Infection with sucrose-purified RSV (pRSV) produced a time-dependent increase in the transcriptional initiation rate of the IL-8 gene. Transient transfection of the human IL-8 promoter mutated in the binding site fur nuclear factor-κB (NF-κB) demonstrated that this sequence was essential for pRSV-activated transcription. Gel mobility shift assays demonstrated pRSV induction of sequence-specific binding complexes; these complexes were supershifted only by antibodies directed to the potent NF-κB transactivating subunit RelA. Both Western immunoblot and indirect immunofluorescence assays showed that cytoplasmic RelA in uninfected cells became localized to the nucleus after pRSV infection. RelA activation requires replicating RSV, because neither conditioned medium nor UV-inactivated pRSV was able to stimulate its translocation. We conclude that RelA undergoes changes in subcellular distribution in airway epithelial cells upon pRSV infection. The ability of replicating RSV to activate RelA translocation may play an important rule in activating IL-8 and other inflammatory gene products necessary for airway mucosal inflammation seen in RSV disease.

UR - http://www.scopus.com/inward/record.url?scp=0029911395&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029911395&partnerID=8YFLogxK

M3 - Article

C2 - 8971006

AN - SCOPUS:0029911395

VL - 70

SP - 8773

EP - 8781

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 12

ER -