Analysis of porcine differential gene expression following challenge with Salmonella enterica serovar Choleraesuis using suppression subtractive hybridization.
Swine-adapted Salmonella enterica subsp. enterica serovar Choleraesuis (S. Choleraesuis) is the pathogen most frequently isolated from diseased pigs and may affect host gene expression in a species-specific manner. To characterize the porcine transcriptional response to S. Choleraesuis infection, the mRNA profiles from the mesenteric lymph nodes of three non-infected and three experimentally infected pigs at 24 h post-inoculation were analyzed by suppression subtractive hybridization (SSH). Forty-four up-regulated and 44 down-regulated genes were revealed by differential cDNA screening of 384 forward and 288 reverse subtracted cDNA clones. The DNA sequence of the cDNA clones identified genes with a role in a variety of cellular functions as well as gene products of unknown function. Seven up-regulated genes (CXCL10, CXCR4, SDCBP, DNAJA1, HSPH1, HSP90 and ANXA5) and two functionally related genes (HSP70 and DNAJA4:pDJA1) were selected for further analysis based on their predicted roles in infection and immunity. Real-time RT-PCR was performed using RNA collected from a time course of infection spanning from the acute phase (8 h) to the chronic phase (21 days) to confirm and quantitate the up-regulation of the SSH-enriched genes. Correlating with the clinical signs of infection (fever, diarrhea and lethargy), the most dramatic induction of gene expression for all nine genes occurred at 48 h post-inoculation. This investigation further defines the porcine response to a host-adapted strain of Salmonella by revealing the differential expression of genes with a role in a variety of host cellular functions including innate immunity and cytoskeleton regulation.
Uthe
Jolita J
JJ
Pre-harvest Food Safety and Enteric Diseases Research Unit, USDA, ARS, National Animal Disease Center, Ames, IA 50010, USA.
Stabel
Thomas J
TJ
Zhao
Shu-hong
SH
Tuggle
Christopher K
CK
Bearson
Shawn M D
SM
D000818:Animals; D017304:Annexin A5 / Q000096:biosynthesis / Q000235:genetics; D019743:Chemokines, CXC / Q000096:biosynthesis / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry; D005260:Female; D020869:Gene Expression Profiling / Q000662:veterinary; D005786:Gene Expression Regulation / Q000276:immunology / Q000502:physiology*; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics; D008198:Lymph Nodes / Q000382:microbiology / Q000503:physiopathology; D008297:Male; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D012481:Salmonella Infections, Animal / Q000235:genetics* / Q000276:immunology / Q000382:microbiology; D019779:Salmonella enterica / Q000502:physiology*; D013552:Swine; D013553:Swine Diseases / Q000235:genetics* / Q000276:immunology / Q000382:microbiology; D053676:Syntenins / Q000096:biosynthesis / Q000235:genetics; D013997:Time Factors; D015854:Up-Regulation
D017304:Annexin A5; D019743:Chemokines, CXC; D017931:DNA Primers; D006360:Heat-Shock Proteins; C109621:SDCBP protein, human; D053676:Syntenins