Coronavirus: proteomics analysis of chicken kidney tissue infected with variant 2 (IS-1494)-like avian infectious bronchitis virus

Abstract

Avian infectious bronchitis virus is one of the most important gammacoronaviruses, which causes a highly contagious disease. In this study, we investigated changes in the proteome of kidney tissue of specific-pathogen-free (SPF) chickens that were infected with an isolate of the nephrotropic variant 2 genotype (IS/1494/06) of avian coronavirus. Twenty 1-day-old SPF White Leghorn chickens were randomly divided into two groups, each comprising 10 chickens, which were kept in separate positive-pressure isolators. Chickens in group A served as a virus-free control group up to the end of the experiment, whereas chickens in group B were inoculated with 0.1 ml of 10 4.5 EID 50 of the IBV/chicken/Iran/UTIVO-C/2014 isolate of IBV, and kidney tissue samples were collected at 2 and 7 days post-inoculation (dpi) from both groups. Sequencing of five protein spots at 2 dpi and 22 spots at 7 dpi that showed differential expression by two-dimensional electrophoresis (2DE) along with fold change greater than 2 was done by MS-MALDI/TOF/TOF. Furthermore, the corresponding protein-protein interaction (PPI) networks at 2 and 7 dpi were identified to develop a detailed understanding of the mechanism of molecular pathogenesis. Topological graph analysis of this undirected PPI network revealed the effect of 10 genes in the 2 dpi PPI network and nine genes in the 7 dpi PPI network during virus pathogenesis. Proteins that were found by 2DE analysis and MS/TOF-TOF mass spectrometry to be down-or upregulated were subjected to PPI network analysis to identify interactions with other cellular components. The results show that cellular metabolism was altered due to viral infection. Additionally, multifunctional heat shock proteins with a significant role in host cell survival may be employed circuitously by the virus to reach its target. The data from this study suggest that the process of pathogenesis that occurs during avian coronavirus infection involves the regulation of vital cellular processes and the gradual disruption of critical cellular functions.

Histopathological study and molecular characterization of Infectious Bronchitis virus circulating in broilers in Basrah governorate, Iraq

“…Nucleotide sequence identities among groups I, II, and III were found to be 99.68%-100%, 99.36%-100%, and 96.42%-100%, respectively, according to sequence analyses Group I (variant2) was the dominant IBV genotype. Variant 2-like strain IS/1494 which detected in our study was characterized with respiratory and renal damages that combined with clinical signs and postmortem lesions of trachea and kidney of infected broilers; such result was related to Abdollahi et al (2021).…”

Analysis of Immune Response of Chicken Primary Cells by Infection With Korean IBV Strain

“…As the rst line of defense, innate immunity is activated within two days after antigenic stimulation. At this initial stage of IBV infection, cytokines and chemokines are produced and co-stimulatory pathways are activated [3,16,17]. Then we speculated the transcriptome analysis of infected kidney tissue by an IBV strain with renal tropism might be helpful to understand the pathogenesis of IB.…”

Evaluation of viral load and transcriptome changes in tracheal tissue of two hybrids of commercial broiler chickens infected with avian infectious bronchitis virus: a comparative study

“…To calculate the viral load, cDNAs from individual tracheal samples in each subgroup were used to amplify a conserved sequence of the IBV genome within the 5’ untranslated region (UTR) by real-time PCR. The forward primer 5′-GCTTTTGAGCCTAGCGTT-3′, located at nucleotide position 391–408 of the genome sequence of IBV strain M41, the reverse primer 5′-GCCATGTTGTCACTGTCTATTG-3′, located at nucleotide position 512–533, and a TaqMan dual-labeled probe (IBV5'G probe 5'-FAMCACCACCAGAACCTGTCACCTC-BHQ1-3') located at nucleotide position 473–494 were designed to amplify and detect a 143-bp fragment of the 5'-UTR [ 20 , 29 ]. The 20-μL reaction mixture contained 2 μL of 10x PCR buffer (Sinaclon), 1 μL of dNTP mix (Sinaclon), 0.8 μL of 50 mM MgCl 2 (Sinaclon), 0.2 μL of CinnaGen Taq DNA polymerase (Sinaclon), 5 μL of template cDNA, 0.4 µL of forward and 0.4 µL of reverse primer at a concentration of 20 μm, 0.4 µL of probe at a concentration of 20 μm, and 9.8 µL of RNase-free water.…”