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Expression of Toll-like receptors and downstream genes in lipopolysaccharide-induced porcine alveolar macrophages.

Authors :
Islam MA
Cinar MU
Uddin MJ
Tholen E
Tesfaye D
Looft C
Schellander K
Source :
Veterinary immunology and immunopathology [Vet Immunol Immunopathol] 2012 Mar 15; Vol. 146 (1), pp. 62-73. Date of Electronic Publication: 2012 Feb 06.
Publication Year :
2012

Abstract

The aim of the present study was to determine the age-related kinetic changes of Toll-like receptors (TLRs) and downstream genes expression, and secretion of cytokine in lipopolysaccharide (LPS) stimulated porcine alveolar macrophages (AM). For this purpose, AMs were isolated from 5-day-old newborn piglets and 120-day-old young pigs. mRNA expression and cytokine measurement was determined by quantitative real-time PCR and ELISA, respectively. First, AMs were incubated for 24 h in the absence or presence of increasing concentrations of LPS. Results showed the up-regulation of TLRs 2, 4, 5 and 9 mRNA from all concentrations of LPS used, as compared to non-stimulated cells, and TLR4 was the highest expression in both ages (P<0.05). Furthermore, quantitative analysis demonstrated increased expression of mRNAs encoding TLRs 2, 4, 5 and 9, LBP, CD14, MD2, MyD88, IRAK4 and TRAF6 in both ages in a time-dependant manner (P<0.05). Overall, LPS inducible mRNA for TLR4, LBP, CD14 and MyD88 had higher expression in newborn piglets compared with those of young pigs (P<0.05). The level of cytokine protein IL6 and TNFα in supernatant fluid significantly varied with time of incubation and age of animals. Their concentration increased immediately at 1 h after LPS stimulation and remained significantly higher up to 48 h in both ages. Production of pro-inflammatory cytokine protein IL6 and TNFα in supernatant was significantly higher in young pigs than those of piglets. This study suggests that differential age-related changes in the expression of TLRs and downstream genes, and pro-inflammatory cytokine could contribute to a different age-related innate immune response during pulmonary infection. Further investigation is warranted to determine the precise effects of LPS on porcine AMs by means of a functional study across a wider age range.<br /> (Copyright © 2012 Elsevier B.V. All rights reserved.)

Details

Language :
English
ISSN :
1873-2534
Volume :
146
Issue :
1
Database :
MEDLINE
Journal :
Veterinary immunology and immunopathology
Publication Type :
Academic Journal
Accession number :
22365308
Full Text :
https://doi.org/10.1016/j.vetimm.2012.02.001