Changes induced by bovine herpesvirus types 1 and 5 infections in toll- like receptors expression patterns on bovine immune cells

MARIN, M; PÉREZ, S; LEUNDA, M; MOORE, D; ODEÓN, A

Lima

Congreso; X Congreso de la Asociación Latinoamericana de Inmunología (ALAI); 2012

Toll like receptors (TLRs) expression in bovine tissues has been described. However, their association with Bovine Herpesvirus types 1 and 5 (BoHV-1 and 5) infections has not been analyzed. The aim of the present study was to investigate the changes in TLR3, 7, 8 and 9 expression levels on BoHV-1- and 5-infected bovine immune cells. PBMC from serum-negative calves (6 × 106 cell/ml) were infected for 24 h with BoHV-1 or BoHV-5 strains (MOI 0.1). Uninfected PBMC were used as control group. Total RNA from PBMC was isolated using Trizol andtreated with DNase I. cDNA was synthesized from 1 µg of RNA. Primers and probes were designed for bovine TLR3, 7, 8 and 9 using the Primer Premier software and GAPDH was used as housekeeping gene. Real time RT-PCR reactions contained 800 nM specific primers, 200 nM probe, 1x PCR Mastermix (TaqMan Universal PCR Mastermix, Applied Biosystems) and 1 µl of cDNA. Quantification of TLR mRNA levels and statistical significance were analysed by the Relative Expression Software Tool. All TLRs analyzed were expressed in uninfected PBMC. Up-regulation of TLR7 following infection by BoHV-1 or BoHV-5 and up-regulation of TLR9 following infection by BoHV-5 were the most striking findings. Although statistically significant differences were not detected in TLR3 expression levels and TLR9 expression in BoHV-1-infected cells, a slight induction of their expression was detected, which might be relevant from a biological point of view. This work reports for the first time the involvement of, at least TLR7 and TLR9, in the recognition of BoHV.