Journal Article

Antiviral activity of bovine type III interferon against bovine viral diarrhea virus is greatly reduced in bovine turbinate cells due to limited expression of IFN lambda receptor 1 (IL-28Rα)

Medina, Gisselle N., Dassanayake, Rohana P., Attreed, Sarah, Sacco, Randy E., Falkenberg, Shollie M., Bickel, Kathryn A., Menghwar, Harish, Casas, Eduardo, De Los Santos, Teresa, Kanipe, Carly, Ma, Hao, Rodriguez-Calzada, Monica, Diaz-San Segunda, Fayna, Holthausen, David J.

2024

Overview

The antiviral activity of recombinant bovine interferon lambda 3 (bovIFN-λ3) against bovine viral diarrhea virus (BVDV) has been demonstrated in Madin-Darby bovine kidney cells (MDBK) and in cattle. However, anti-BVDV activity of bovIFN-λ3 has not been studied in bovine respiratory tract epithelial cells, supposedly a primary target of BVDV infection when entering the host by the oronasal route. Here we investigated the anti-BVDV activity of bovIFN-λ3 in bovine turbinate-derived primary epithelial cells (BTu) using BVDV infection and immunoperoxidase staining, TCID , RT-qPCR, DNA and transcriptome sequencing, and transfection with plasmids containing the two subunits, IL-28Rα and IL-10Rβ that constitute the bovIFN-λ3 receptor. Our immunoperoxidase staining, RT-qPCR, and TCID results show that while BVDV was successfully cleared in MDBK cells treated with bovIFN-λ3 and bovIFN-α, only the latter, bovIFN-α, cleared BVDV in BTu cells. Preincubation of MDBK cells with bovIFN-λ3 before BVDV infection was needed to induce optimal antiviral state. Both cell types displayed intact type I and III IFN signaling pathways and expressed similar levels of IL-10Rβ subunit of the type III IFN receptor. Sequencing of PCR amplicon of the IL-28Rα subunit revealed intact transmembrane domain and lack of single nucleotide polymorphisms (SNPs) in BTu cells. However, RT-qPCR and transcriptomic analyses showed a lower expression of IL-28Rα transcripts in BTu cells as compared to MDBK cells. Interestingly, transfection of BTu cells with a plasmid encoding IL-28Rα subunit, but not IL-10Rβ subunit, established the bovIFN-λ3 sensitivity showing similar anti-BVDV activity to the response in MDBK cells. Our results demonstrate that the sensitivity of cells to bovIFN-λ3 depends not only on the quality but also of the quantity of the IL-28Rα subunit of the heterodimeric receptor. A reduction in IL-28Rα transcript expression was detected in BTu as compared to MDBK cells, despite the absence of spliced variants or SNPs. The establishment of bovIFN-λ3 induced anti-BVDV activity in BTu cells transfected with an IL-28Rα plasmid suggests that the level of expression of this receptor subunit is crucial for the specific antiviral activity of type III IFN in these cells.

• Publisher: Frontiers Media SA,Frontiers Media S.A
• Subject: Animals / Antiviral activity / Antiviral Agents - pharmacology / antiviral properties / Antiviral state / Bone / bovine turbinate primary epithelial cells / bovine type III interferon / bovine viral diarrhea virus / Bovine viral diarrhea virus 1 / Bovine Virus Diarrhea-Mucosal Disease - immunology / Bovine Virus Diarrhea-Mucosal Disease - virology / Cattle / Cell culture / Cell death / Cell Line / Cytokines / Dendritic cells / Diarrhea / Diarrhea Viruses, Bovine Viral - immunology / Diarrhea Viruses, Bovine Viral - physiology / DNA sequencing / domain / Epithelial cells / Epithelial Cells - immunology / Epithelial Cells - metabolism / Epithelial Cells - virology / epithelium / Genes / IFN-λ3 / IL-10Rβ / IL-28Rα / Immunology / Interferon / Interferon Lambda / interferons / Interferons - immunology / Interferons - metabolism / Interleukin-10 Receptor beta Subunit - genetics / Interleukin-10 Receptor beta Subunit - metabolism / Interleukins - genetics / Interleukins - immunology / Interleukins - metabolism / Interleukins - pharmacology / kidneys / Kinases / Laboratories / Lymphatic system / Monoclonal antibodies / Nose / Plasmids / Proteins / Receptors, Cytokine / Receptors, Interleukin - genetics / Receptors, Interleukin - metabolism / Recombinant Proteins - pharmacology / respiratory system / Respiratory tract / Signal transduction / Single-nucleotide polymorphism / transcriptome / Transcriptomes / Transcriptomics / Transfection / Transmembrane domains / Turbinates - immunology / Turbinates - metabolism / Turbinates - virology / Viral infections / Viruses