title: Differential host gene expression in cells infected with highly pathogenic H5N1 avian influenza viruses.
abstract: In order to understand the molecular mechanisms by which different strains of avian influenza viruses overcome host response in birds, we used a complete chicken genome microarray to compare early gene expression levels in chicken embryo fibroblasts (CEF) infected with two avian influenza viruses (AIV), A/CK/Hong Kong/220/97 and A/Egret/Hong Kong/757.2/02, with different replication characteristics. Gene ontology revealed that the genes with altered expression are involved in many vital functional classes including protein metabolism, translation, transcription, host defense/immune response, ubiquitination and the cell cycle. Among the immune-related genes, MEK2, MHC class I, PDCD10 and Bcl-3 were selected for further expression analysis at 24 hpi using semi-quantitive RT-PCR. Infection of CEF with A/Egret/Hong Kong/757.2/02 resulted in a marked repression of MEK2 and MHC class I gene expression levels. Infection of CEF with A/CK/Hong Kong/220/97 induced an increase of MEK2 and a decrease in PDCD10 and Bcl-3 expression levels. The expression levels of alpha interferon (IFN-alpha), myxovirus resistance 1 (Mx1) and interleukin-8 (IL-8) were also analyzed at 24 hpi, showing higher expression levels of all of these genes after infection with A/CK/Hong Kong/220/97 compared to A/Egret/Hong Kong/757.2/02. In addition, comparison of the NS1 sequences of the viruses revealed amino acid differences that may explain in part the differences in IFN-alpha expression observed. Microarray gene expression analysis has proven to be a useful tool on providing important insights into how different AIVs affect host gene expression and how AIVs may use different strategies to evade host response and replicate in host cells.
authors.affiliation: Southeast Poultry Research Laboratory, Agricultural Research Service, United States Department of Agriculture, Athens, GA 30605, USA.
authors.forename: Luciana
authors.initials: L
authors.lastname: Sarmento
authors.forename: Claudio L
authors.initials: CL
authors.lastname: Afonso
authors.forename: Carlos
authors.initials: C
authors.lastname: Estevez
authors.forename: Jamie
authors.initials: J
authors.lastname: Wasilenko
authors.forename: Mary
authors.initials: M
authors.lastname: Pantin-Jackwood
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002642:Chick Embryo; D002645:Chickens; D005347:Fibroblasts / Q000821:virology; D015967:Gene Expression Regulation, Viral; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics / Q000254:growth & development / Q000276:immunology / Q000502:physiology*; D005585:Influenza in Birds / Q000235:genetics* / Q000276:immunology* / Q000821:virology; D016898:Interferon-alpha / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D020411:Oligonucleotide Array Sequence Analysis; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013047:Specific Pathogen-Free Organisms; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D016898:Interferon-alpha; D012367:RNA, Viral; D017361:Viral Nonstructural Proteins