Unknown,Transcriptomics,Genomics,Proteomics

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LPAIV in chicken

ABSTRACT: Animal experiments
Experiment : Chickens of group 1 were inoculated with 0.2 ml (2*105 EID50) of the H7N1 HPAIV strain, equally divided between the intranasal and intratracheal route. In the same way, animals of group 2 received 2*105 EID50 of the H7N1 LPAIV strain. Sampling: Six chickens from each group were sacrificed just before infection (controls) and at 4, 8, 16 and 24 h.p.i. From all sacrificed chickens, gross pathology of the organs was studied. Lung, trachea, ileum and brains were collected. For RNA isolation, organs were snap-frozen in liquid nitrogen and stored at -80C until use. For immonohistochemistry, samples were fixed immediately by immersion in a zinc salt-based fixative, containing 0.5 % zinc chloride, 0.5% zinc acetate in 0.1 M Tris base buffer containing 0.05 % calcium acetate, pH 7.4 for 48 h. After fixation, the tissues were dehydrated and embedded in paraffin wax. All HPAIV experiments were performed in Biosafety Level 3 facilities.

ORGANISM(S): Gallus gallus

SUBMITTER: Jacob Post

PROVIDER: E-MEXP-3109 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Highly pathogenic avian influenza virus (HPAIV) is a permanent threat due to its capacity to cross species barriers and generate severe infections and high mortality in humans. Recent findings have highlighted the potential role of PB1-F2, a small accessory influenza protein, in the pathogenesis process mediated by HPAIV in mammals. In this study, using a recombinant H5N1 HPAIV (wt) and its PB1-F2-deleted mutant (M-NM-^TF2), we studied the effects of PB1-F2 in a chicken model. Unexpectedly, when using low inoculation dose we observed that the wt-infected chickens had a higher survival rate than the M-NM-^TF2-infected chickens, a feature that contrasts with what is usually observed in mammals. High inoculation dose had similar mortality rate for both viruses, and comparison of the bio-distribution of the two viruses indicated that the expression of PB1-F2 allows a better spreading of the virus within chicken embryos. Transcriptomic profiles of lungs and blood cells were characterized at two days post-infection in chickens inoculated with the wild type (wt) or the M-NM-^TF2 mutant viruses. In lungs, the expression of PB1-F2 during the infection induced pathways related to calcium signaling and repressed a large panel of immunological functions. In blood cells, PB1-F2 was associated to a gene signature specific for mitochondrial dysfunction and down-modulated leucocytes activation. Finally we compared the effect of PB1-F2 in lungs of chickens and mice. We identified that gene signature associated to tissue damages is a PB1-F2 feature shared by the two species; by contrast, the early inhibition of immune response mediated by PB1-F2 observed in chickens is not seen in mice. In summary, our data suggest that PB1-F2 expression deeply affect the immune host response in chickens in a way that may attenuate pathogenicity, a feature differing from what was previously observed in mammal species. Three-condition experiment, virus-infected (wt or M-NM-^TF2) vs. Mock-infected chickens. Biological replicates: 2x5 control replicates, 5 wt replicates, 5 M-NM-^TF2 replicates.

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LPAIV in chicken

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