cDNA microarray technology was applied to time course analysis of differentially expressed genes in A549 cells following human respiratory syncytial virus (HRSV) infection. Both up- and down-regulation of cellular genes were observed in a time-dependent manner. However, gene up-regulation prevailed over gene down-regulation. Virus infectivity was required as UV-inactivated virus failed to up-regulate/down-regulate those genes. At early times post-infection (0–6 h p.i.) 85 genes were up-regulated. Some of those genes were involved in cell growth/proliferation, cellular protein metabolism and cytoskeleton organization. Among the most strongly up-regulated genes at that time were the urokinase plasminogen activator (PLAU) and its receptor (PLAUR), a pleiotropic system involved in many biological processes, including chemotaxis and inflammation. Functionally related genes encoding the α- and β-chains of several integrins were also up-regulated within the first 12 h of infection. Genes up-regulated between 6 and 12 h p.i. included interferon-stimulated genes (ISGs), genes related to oxidative stress and genes of the non-canonical NF-κB pathway. At later times, genes involved in the immune response became predominant among the up-regulated genes, most of them being ISGs. Different up-regulation kinetics of cytokine and cytokine-signalling-related genes were also observed. These results highlight the dynamic interplay between the virus and the host cell and provide a general picture of changes in cellular gene expression along the HRSV replicative cycle.
AlexanderW. S.,
HiltonD. J.2004; The role of suppressors of cytokine signaling (SOCS) proteins in regulation of the immune response. Annu Rev Immunol 22:503–529[CrossRef]
Al-ShahrourF.,
Diaz-UriarteR.,
DopazoJ.2004; FatiGO: a web tool for finding significant associations of Gene Ontology terms with groups of genes. Bioinformatics 20:578–580[CrossRef]
BorghesiL. A.,
LynesM. A.1996; Stress proteins as agents of immunological change: some lessons from metallothionein. Cell Stress Chaperones 1:99–108[CrossRef]
BukreyevA.,
BelyakovI. M.,
PrinceG. A.,
YimK. C.,
HarrisK. K.,
BerzofskyJ. A.,
CollinsP. L.2005; Expression of interleukin-4 by recombinant respiratory syncytial virus is associated with accelerated inflammation and a nonfunctional cytotoxic T-lymphocyte response following primary infection but not following challenge with wild-type virus. J Virol 79:9515–9526[CrossRef]
BurkeE.,
DupuyL.,
WallC.,
BarikS.1998; Role of cellular actin in the gene expression and morphogenesis of human respiratory syncytial virus. Virology 252:137–148[CrossRef]
CollinsP. L.,
ChanockR. M.,
MurphyB. R.2001; Respiratory syncytial virus. In Fields Virology, 4th edn. pp 1443–1485 Edited by
KnipeD. M.,
HowleyP. M.
Philadelphia: Lippincott Williams & Wilkins;
FalseyA. R.,
HennesseyP. A.,
FormicaM. A.,
CoxC.,
WalshE. E.2005; Respiratory syncytial virus infection in elderly and high-risk adults. N Engl J Med 352:1749–1759[CrossRef]
FawazL. M.,
Sharif-AskariE.,
MenezesJ.1999; Up-regulation of NK cytotoxic activity via IL-15 induction by different viruses: a comparative study. J Immunol 163:4473–4480
Garcia-BarrenoB.,
JorcanoJ. L.,
AukenbauerT.,
Lopez-GalindezC.,
MeleroJ. A.1988; Participation of cytoskeletal intermediate filaments in the infectious cycle of human respiratory syncytial virus (RSV). Virus Res 9:307–321[CrossRef]
Garcia-BarrenoB.,
PalomoC.,
PenasC.,
DelgadoT.,
Perez-BrenaP.,
MeleroJ. A.1989; Marked differences in the antigenic structure of human respiratory syncytial virus F and G glycoproteins. J Virol 63:925–932
GarofaloR.,
MeiF.,
EspejoR.,
YeG.,
HaeberleH.,
BaronS.,
OgraP. L.,
ReyesV. E.1996; Respiratory syncytial virus infection of human respiratory epithelial cells up-regulates class I MHC expression through the induction of IFN-beta and IL-1 alpha. J Immunol 157:2506–2513
GowerT. L.,
PasteyM. K.,
PeeplesM. E.,
CollinsP. L.,
McCurdyL. H.,
HartT. K.,
GuthA.,
JohnsonT. R.,
GrahamB. S.2005; RhoA signaling is required for respiratory syncytial virus-induced syncytium formation and filamentous virion morphology. J Virol 79:5326–5336[CrossRef]
GuoJ.,
PetersK. L.,
SenG. C.2000; Induction of the human protein P56 by interferon, double-stranded RNA, or virus infection. Virology 267:209–219[CrossRef]
HerreroJ.,
Al-ShahrourF.,
Diaz-UriarteR.,
MateosA.,
VaquerizasJ. M.,
SantoyoJ.,
DopazoJ.2003a; gepas: a web-based resource for microarray gene expression data analysis. Nucleic Acids Res 31:3461–3467[CrossRef]
HornungV.,
SchlenderJ.,
Guenthner-BillerM.,
RothenfusserS.,
EndresS.,
ConzelmannK. K.,
HartmannG.2004; Replication-dependent potent IFN-alpha induction in human plasmacytoid dendritic cells by a single-stranded RNA virus. J Immunol 173:5935–5943[CrossRef]
JohnsonT. R.,
ParkerR. A.,
JohnsonJ. E.,
GrahamB. S.2003; IL-13 is sufficient for respiratory syncytial virus G glycoprotein-induced eosinophilia after respiratory syncytial virus challenge. J Immunol 170:2037–2045[CrossRef]
KallewaardN. L.,
BowenA. L.,
CroweJ. E., Jr.
2005; Cooperativity of actin and microtubule elements during replication of respiratory syncytial virus. Virology 331:73–81[CrossRef]
MartínezI.,
DopazoJ.,
MeleroJ. A.1997; Antigenic structure of the human respiratory syncytial virus G glycoprotein and relevance of hypermutation events for the generation of antigenic variants. J Gen Virol 78:2419–2429
Martinez-DelgadoB.,
MelendezB.,
CuadrosM.,
AlvarezJ.,
CastrilloJ. M.,
Ruiz De La ParteA.,
MollejoM.,
BellasC.,
DiazR.other authors2004; Expression profiling of T-cell lymphomas differentiates peripheral and lymphoblastic lymphomas and defines survival related genes. Clin Cancer Res 10:4971–4982[CrossRef]
MbiguinoA.,
MenezesJ.1991; Purification of human respiratory syncytial virus: superiority of sucrose gradient over percoll, renografin, and metrizamide gradients. J Virol Methods 31:161–170[CrossRef]
OkumaY.,
O'ConnorA. E.,
MuirJ. A.,
StantonP. G.,
de KretserD. M.,
HedgerM. P.2005; Regulation of activin A and inhibin B secretion by inflammatory mediators in adult rat Sertoli cell cultures. J Endocrinol 187:125–134[CrossRef]
PolackF. P.,
TengM. N.,
CollinsP. L.,
PrinceG. A.,
ExnerM.,
RegeleH.,
LirmanD. D.,
RaboldR.,
HoffmanS. J.other authors2002; A role for immune complexes in enhanced respiratory syncytial virus disease. J Exp Med 196:859–865[CrossRef]
RamaswamyM.,
ShiL.,
MonickM. M.,
HunninghakeG. W.,
LookD. C.2004; Specific inhibition of type I interferon signal transduction by respiratory syncytial virus. Am J Respir Cell Mol Biol 30:893–900[CrossRef]
RamaswamyM.,
ShiL.,
VargaS. M.,
BarikS.,
BehlkeM. A.,
LookD. C.2006; Respiratory syncytial virus nonstructural protein 2 specifically inhibits type I interferon signal transduction. Virology 344:328–339[CrossRef]
ResnatiM.,
PallaviciniI.,
WangJ. M.,
OppenheimJ.,
SerhanC. N.,
RomanoM.,
BlasiF.2002; The fibrinolytic receptor for urokinase activates the G protein-coupled chemotactic receptor FPRL1/LXA4R. Proc Natl Acad Sci U S A 99:1359–1364[CrossRef]
ShoudaT.,
YoshidaT.,
HanadaT.,
WakiokaT.,
OishiM.,
MiyoshiK.,
KomiyaS.,
KosaiK.,
HanakawaY.other authors2001; Induction of the cytokine signal regulator SOCS3/CIS3 as a therapeutic strategy for treating inflammatory arthritis. J Clin Invest 108:1781–1788[CrossRef]
SpannK. M.,
TranK. C.,
CollinsP. L.2005; Effects of nonstructural proteins NS1 and NS2 of human respiratory syncytial virus on interferon regulatory factor 3, NF-kappaB, and proinflammatory cytokines. J Virol 79:5353–5362[CrossRef]
TangY. W.,
GrahamB. S.1994; Anti-IL-4 treatment at immunization modulates cytokine expression, reduces illness, and increases cytotoxic T lymphocyte activity in mice challenged with respiratory syncytial virus. J Clin Invest 94:1953–1958[CrossRef]
TraceyL.,
SpiteriI.,
OrtizP.,
LawlerM.,
PirisM. A.,
VilluendasR.2004; Transcriptional response of T cells to IFN-alpha: changes induced in IFN-alpha-sensitive and resistant cutaneous T cell lymphoma. J Interferon Cytokine Res 24:185–195[CrossRef]
TusherV. G.,
TibshiraniR.,
ChuG.2001; Significance analysis of microarrays applied to the ionizing radiation response. Proc Natl Acad Sci U S A 98:5116–5121[CrossRef]
UlloaL.,
SerraR.,
AsenjoA.,
VillanuevaN.1998; Interactions between cellular actin and human respiratory syncytial virus (HRSV). Virus Res 53:13–25[CrossRef]
YangY. H.,
DudoitS.,
LuuP.,
LinD. M.,
PengV.,
NgaiJ.,
SpeedT. P.2002; Normalization for cDNA microarray data: a robust composite method addressing single and multiple slide systematic variation. Nucleic Acids Res 30:e15[CrossRef]