Gene expression pattern in swine neutrophils after lipopolysaccharide exposure: a time course comparison
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Author
Sanz-Santos, Gema
Jiménez-Marín, Ángeles
Bautista, Rocío
Fernández, Noé
Claros, Gonzalo M.
Garrido, Juan J.
Publisher
BioMed CentralDate
2011Subject
Innate immune responseBacterial infection
Swine
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Background: Experimental exposure of swine neutrophils to bacterial lipopolysaccharide (LPS) represents a model
to study the innate immune response during bacterial infection. Neutrophils can effectively limit the infection by
secreting lipid mediators, antimicrobial molecules and a combination of reactive oxygen species (ROS) without new
synthesis of proteins. However, it is known that neutrophils can modify the gene expression after LPS exposure. We
performed microarray gene expression analysis in order to elucidate the less known transcriptional response of
neutrophils during infection.
Methods: Blood samples were collected from four healthy Iberian pigs and neutrophils were isolated and incubated
during 6, 9 and 18 hrs in presence or absence of lipopolysaccharide (LPS) from Salmonella enterica serovar Typhimurium.
RNA was isolated and hybridized to Affymetrix Porcine GeneChip®. Microarray data were normalized using Robust
Microarray Analysis (RMA) and then, differential expression was obtained by an analysis of variance (ANOVA).
Results: ANOVA data analysis showed that the number of differentially expressed genes (DEG) after LPS treatment vary
with time. The highest transcriptional response occurred at 9 hr post LPS stimulation with 1494 DEG whereas at 6 and
18 hr showed 125 and 108 DEG, respectively. Three different gene expression tendencies were observed: genes in
cluster 1 showed a tendency toward up-regulation; cluster 2 genes showing a tendency for down-regulation at 9 hr;
and cluster 3 genes were up-regulated at 9 hr post LPS stimulation. Ingenuity Pathway Analysis revealed a delay of
neutrophil apoptosis at 9 hr. Many genes controlling biological functions were altered with time including those
controlling metabolism and cell organization, ubiquitination, adhesion, movement or inflammatory response.
Conclusions: LPS stimulation alters the transcriptional pattern in neutrophils and the present results show that the
robust transcriptional potential of neutrophils under infection conditions, indicating that active regulation of gene
expression plays a major role in the neutrophil-mediated- innate immune response