Gene expression profiling of bovine macrophages in response to Escherichia coli O157:H7 lipopolysaccharide.
The aim of this study was to identify changes in bovine macrophage gene expression in response to treatment with Escherichia coli 0157:H7 lipopolysaccharide (LPS), utilizing a human gene microarray. Bovine cDNA from control and LPS-treated primary macrophages hybridized to greater than 5644 (79.8%) of the non-control gene targets on a commercially available microarray containing greater than 7075 targets (Incyte Genomics, St. Louis, MO). Of these target sequences, 44 were differentially expressed upon exposure to LPS, including 18 genes not previously reported to exist in cattle. These included a pentaxin-related gene, CASP8, TNF-induced genes, interferon-induced genes, and inhibitors of apoptosis. Using the human microarray, cDNA from bovine LPS-treated and control macrophages consistently hybridized to targets known to be expressed constitutively by macrophages, as expected given the predicted cDNA sequence homology. That this human system was accurately estimating levels of bovine transcripts was further verified by real-time quantitative reverse transcriptase polymerase chain reaction (RTQ-PCR) using bovine-specific primers. This first report of bovine-human cross-species expression profiling by microarray hybridization demonstrates the utility of this technique in bovine gene expression and discovery.
Chitko-McKown
Carol G
CG
USDA, ARS, Roman L. Hruska US Meat Animal Research Center (MARC), State Spur 18D, P.O. Box 166, Clay Center, Nebraska 68933-0166, USA.
Fox
James M
JM
Miller
Laura C
LC
Heaton
Michael P
MP
Bono
James L
JL
Keen
James E
JE
Grosse
William M
WM
Laegreid
William W
WW
D000818:Animals; D002417:Cattle / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D002418:Cattle Diseases / Q000382:microbiology*; D004927:Escherichia coli Infections / Q000276:immunology / Q000382:microbiology / Q000662:veterinary*; D019453:Escherichia coli O157 / Q000276:immunology*; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008264:Macrophages / Q000187:drug effects / Q000276:immunology / Q000502:physiology*; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D016415:Sequence Alignment
D008070:Lipopolysaccharides; D012313:RNA