Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of pig alveolar macrophages infected with Porcine Respiratory Reproductive Syndrome Virus

ABSTRACT: A primary culture of Porcine Alveolar Macrophages (PAM) from six animals was split in two: one was infected at a multiplicity of infection (MOI) of 10 with a 13th passage of PRRSV Lelystad virus. The other culture was maintained as control. The percentage of infected cells ranged between 60 and 70% for all batches. Cells were collected 1, 3, 6, 9 and 12 hours Post Infection in Trizolfor RNA extraction. Total RNA extraction from PAM was performed with Trizol following standard instructions and clean-up was done using RNeasy columns. RNA quality was assessed by microcapillary electrophoresis on an Agilent 2001 Bioanalyzer with RNA 6000 Nanochips. RNA was quantified by spectrophotometry. Reverse transcription (RT) of 20 ÃÂµg total RNA and synthesis of biotin-labeled cRNA with one round of amplification were carried out following the standard Affymetrix one-cycle protocol according to the manufacturer's instructions. Three samples each from control and infected cells were pooled (resulting in 2 control and 2 infected samples for analysis at each time point), and their transcriptional profiles were assessed on GeneChip Porcine Genome Arrays. (http://www.affymetrix.com/products/arrays/specific/porcine.affx).

ORGANISM(S): Sus scrofa

DISEASE(S): Control

SUBMITTER: Roberto Malinverni

PROVIDER: E-MEXP-1350 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Genome-wide transcriptional response of primary alveolar macrophages following infection with porcine reproductive and respiratory syndrome virus.

Genini Sem S Delputte Peter L PL Malinverni Roberto R Cecere Maria M Stella Alessandra A Nauwynck Hans J HJ Giuffra Elisabetta E

The Journal of general virology 20081001 Pt 10

Porcine reproductive and respiratory syndrome is a major cause of economic loss for the swine industry worldwide. Porcine reproductive and respiratory syndrome virus (PRRSV) triggers weak and atypical innate immune responses, but key genes and mechanisms by which the virus interferes with the host innate immunity have not yet been elucidated. In this study, genes that control the response of the main target of PRRSV, porcine alveolar macrophages (PAMs), were profiled in vitro with a time-course ...[more]

PMID: 18796724

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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.

Dataset Information

Transcription profiling of pig alveolar macrophages infected with Porcine Respiratory Reproductive Syndrome Virus

Publications

Genome-wide transcriptional response of primary alveolar macrophages following infection with porcine reproductive and respiratory syndrome virus.

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