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Detection of gene expression changes in S. Choleraesuis infected porcine lung using an oligonucleotide microarray

ABSTRACT: Piglets were raised in isolation facilities from 10 days to 7 weeks of age at the National Animal Disease Center. Bacteriological culture of rectal swabs was performed twice to confirm that all pigs were free of Salmonella spp. Three control pigs were necropsied on experimental day -3. On day 0, pigs in the infected group were intranasally challenged with 1 x 109 CFU S. Choleraesuis X3246. Lung samples were aseptically collected from three 24 h and three 48 h post infection pigs and immediately frozen in liquid nitrogen for future mRNA isolation. In the microarray experiment, the nine animals were randomly assigned to three loops so that each loop included one animal from each infection status category. A total of 9 slides (3 slides per loop) were used to obtain a total of 18 measurements from the three control and six infected pigs. A mixed model analyses was also used to identify differentially expressed genes. The mixed model included fixed effects for dye (Cy3 or Cy5), time point (control, 24 hour, and 48 hour infection) as well as random effects for slide and animal. Keywords: repeat sample

ORGANISM(S): Sus scrofa

PROVIDER: GSE2339 | GEO | 2005/03/05

SECONDARY ACCESSION(S): PRJNA91561

REPOSITORIES: GEO

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Project description:A first generation Affymetrix GeneChipÂ® Porcine genome array was used to profile the gene expression in porcine mesenteric lymph nodes over a time course of infection with S. Typhimurium, including the acute (8 hours post inoculation (hpi), 24 hpi, 48 hpi) and chronic (21 days post-inoculation (dpi)) stages of infection. Our objectives were to 1) identify and examine the stereotypical gene expression response within host MLN to S. Typhimurium infection, 2) characterize global host responses by revealing the specific features of the hostâs innate immunity pathways, and 3) explore if and how S. Typhimurium may escape the host immune response and develop into a carrier state. Our study has attempted to investigate the features of host gene expression profiling during S. Typhimurium infection at the acute and chronic infection stages and to explore the mechanism by which S. Typhimurium can escape from the host immune response and develop a carrier state in the host. In conclusion, by using the Affymetrix porcine GeneChip, 848 differentially expressed genes were identified in porcine MLN during infection and several specific features of host response were revealed by gene cluster and pathway analysis. Our data are the first report to investigate global host responses to S. Typhimurium in porcine MLN, and this new study provides data applicable for studying enteric salmonellosis of pigs, as well as humans. Experiment Overall Design: Fifteen piglets from Salmonella spp.-free sows were weaned at 10 days (d) of age, shipped to the National Animal Disease Center, Ames, IA and raised in isolation facilities. To confirm that all piglets were free of Salmonella spp. prior to challenge, bacteriological cultures were performed on rectal swabs twice. Seven week old pigs were randomly divided into 2 groups, 3 non-infected pigs and 12 infected pigs. Three non-infected control pigs were necropsied 2 days prior to experimental infection. On day 0, pigs in the infected groups were intranasally challenged with 1 billion CFU of Salmonella enterica serovar Typhimurium. Three infected pigs were necropsied at 8 hours post-inoculation (hpi), 24 hpi, 48 hpi and 21 days post-inoculation (dpi). Tissue samples from the mesenteric lymph nodes (MLN) were collected and immediately frozen in liquid nitrogen for RNA isolation.

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Detection of gene expression changes in S. Choleraesuis infected porcine lung using an oligonucleotide microarray

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