Analysis of lung transcriptome in calves infected with Bovine Respiratory Syncytial Virus and treated with antiviral and/or cyclooxygenase inhibitor

Abstract

Bovine Respiratory Syncytial virus (BRSV) is one of the major infectious agents in the etiology of the bovine respiratory disease complex. BRSV causes a respiratory syndrome in calves, which is associated with severe bronchiolitis. In this study we describe the effect of treatment with antiviral fusion protein inhibitor (FPI) and ibuprofen, on gene expression in lung tissue of calves infected with BRSV. Calves infected with BRSV are an excellent model of human RSV in infants: we hypothesized that FPI in combination with ibuprofen would provide the best therapeutic intervention for both species. The following experimental treatment groups of BRSV infected calves were used: 1) ibuprofen day 3–10, 2) ibuprofen day 5–10, 3) placebo, 4) FPI day 5–10, 5) FPI and ibuprofen day 5–10, 6) FPI and ibuprofen day 3–10. All calves were infected with BRSV on day 0. Daily clinical evaluation with monitoring of virus shedding by qRT-PCR was conducted. On day10 lung tissue with lesions (LL) and non-lesional (LN) was collected at necropsy, total RNA extracted, and RNA sequencing performed. Differential gene expression analysis was conducted with Gene ontology (GO) and KEGG pathway enrichment analysis. The most significant differential gene expression in BRSV infected lung tissues was observed in the comparison of LL with LN; oxidative stress and cell damage was especially noticeable. Innate and adaptive immune functions were reduced in LL. As expected, combined treatment with FPI and Ibuprofen, when started early, made the most difference in gene expression patterns in comparison with placebo, especially in pathways related to the innate and adaptive immune response in both LL and LN. Ibuprofen, when used alone, negatively affected the antiviral response and caused higher virus loads as shown by increased viral shedding. In contrast, when used with FPI Ibuprofen enhanced the specific antiviral effect of FPI, due to its ability to reduce the damaging effect of prostanoids and oxidative stress.

Integrated Network Analysis to Identify Key Modules and Potential Hub Genes Involved in Bovine Respiratory Disease: A Systems Biology Approach

“…Several functional terms identified in the purple module included “Natural killer cell-mediated cytotoxicity,” “T-cell receptor signaling pathway,” “cellular defense response,” “Chemokine signaling pathway,” and “T-cell activation.” Interestingly, in addition to the antiviral activity of T cells that have been discussed above, the T-cell receptor signaling pathway has also been observed in challenges with Mannheimia haemolytica , which indicates the importance and participation of T cells in responding to different types of pathogens ( Tizioto et al, 2015 ). In agreement with the previous studies, PRF1 ( Johnston et al, 2021a ), LCK ( Smirnova et al, 2009 ), NCR1 ( Osman and Griebel, 2017 ), CCL5 ( N’jai et al, 2013 ), CD8A ( Knapek et al, 2020 ; Lebedev et al, 2021 ), CCR8 ( Lopez et al, 2020 ), CX3CR1 ( Salem et al, 2019 ), and TBX21 hub–hub TF ( Johnston et al, 2019 ) were identified as highly connected genes in the purple module and have been reported as immune-related genes during BRD. For instance, the PRF1 hub–hub gene is an important gene that encodes the perforin-1, which is present in cytotoxic T-lymphocytes (CTLs) and natural killer cells (NK cells) and is involved in cytolysis and the regulation of the immune system.…”

“…Functional terms in the blue module were mostly related to the innate immune system, including “lysosome,” “phagosome,” “Fc gamma R-mediated phagocytosis,” “leukocyte transendothelial migration,” “Toll-like receptor signaling pathway,” “Chemokine signaling pathway,” “neutrophil activation involved in immune response,” “activation of MAPK activity,” and “positive regulation of NF-kappaB import into nucleus.” The leukocyte transendothelial migration is one of the important steps of the innate immune system in triggering the inflammatory immune response and the migration of the first immune response cells such as neutrophils to the sites of infection ( Getter et al, 2019 ; Bakhtiarizadeh et al, 2020 ) which that chemokines control cellular responses at inflammatory sites through this pathway ( Krishnan et al, 2004 ). In agreement with similar BRD studies ( Tizioto et al, 2015 ; Behura et al, 2017 ; Johnston et al, 2021c ; Lebedev et al, 2021 ), the leukocyte transendothelial migration was identified as one of the important pathways of the immune system in the blue module. Additionally, some of the hub–hub genes identified in the blue module have also been reported in the previous BRD studies, including TLR4 ( Scott et al, 2021 ), ANXA5 ( Mitchell et al, 2008 ), C3 , PSEN1 ( Neupane et al, 2018 ), CTSB , CD59 , FTL ( Nilson et al, 2020 ), CAT ( Joshi et al, 2018 ), TLR7 , CD86 ( Palomares et al, 2014 ), ATG7 ( Lipkin et al, 2016 ), and IFNAR1 ( Amat et al, 2019 ).…”

A Genome-Wide Association Study for Resistance to Tropical Theileriosis in Two Bovine Portuguese Autochthonous Breeds

“…Another gene recognized was CMTM7 (KLF-like MARVEL transmembrane domain-containing 7), which belongs to the superfamily encoding chemokine-like factors. A lack of CMTM7 has already been shown to cause a reduction in the innate B-cell population and lead to natural IgM and IL-10 deficiency [43]. IL 10 is one of the interleukins present in the highest concentration when cattle are infected with T. annulata and is essential in the development of the immune response against this agent [44].…”

Sığırların Solunum Sistemi Hastalığı Kompleksi

“…Hastalığın klinik seyri olarak buzağılarda rinitis, laringofarenjit, trakeobronşit ve hastalığın ilerleyen aşamalarında akciğerlerde hırıltılı bir şekilde bronşitis, taşipnö ve öksürük görülebilmektedir. Ayrıca buzağılarda ateş, anoreksi ve depresyon hali de bulunmaktadır [42]. Baker ve ark., [43] BRSV enfeksiyonu bulunan sığırlarda hastalık ilerledikçe dispnönün şiddetinin arttığını, ağzı açık solunumun şekillendiğini ve geçici bir diyarenin oluşabileceğini belirtmişlerdir.…”