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

MARIN M; PÉREZ S; QUINTANA S; LEUNDA M; MOORE P; ODEÓN A

Congreso; X Congreso de la Asociación Latinoamericana de Inmunología; 2012

Asociación Latinoamericana de Inmunología,

Involvement of Toll like receptors (TLR) expression on Bovine Herpesvirus (BoHV) infections has not been analyzed. The aim of this study was to investigate the changes in TLR3, 7, 8 and 9 expression levels on BoHV-infected bovine immune cells. PBMC (6 × 106 cell/ml) were infected for 24 h with BoHV-1 or BoHV-5 (MOI 0.1). Uninfected PBMC were used as control group. RNA from PBMC was isolated using Trizol and cDNA was synthesized from 1 µg of RNA. Primers and probes were designed 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 TaqMan PCR Mastermix and 1 µl of cDNA. Statistical significance were analysed by the Relative Expression Software Tool. All TLR analyzed were expressed in uninfected PBMC. Up-regulation of TLR7 following infection by BoHV-1 or BoHV-5 (3.4 and 5.4 fold increase, respectively) and up-regulation of TLR9 following infection by BoHV-5 (2.7 fold increase) 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 (1.4 fold increase), 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.