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-28Ra)

Authors: Dassanayake, Rohana; MENGHWAR, HARISH - Oak Ridge Institute For Science And Education (ORISE); Mc Mullen, Kathryn; Bickel, Kathryn; Holthausen, David; Ma, Hao; SEGUNDA, FAYNA DIAZ-SAN - Nih, National Institutes Of Allergy And Infectious Diseases; RODRIGUEZ-CALZADA, MONICA - Oak Ridge Institute For Science And Education (ORISE); Medina, Gisselle; Attreed, Sarah; FALKENBERG, SHOLLIE - Auburn University; Kanipe, Carly; Sacco, Randy; De Los Santos, Teresa; Casas, Eduardo

Submitted to: Frontiers in Immunology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/31/2024
Publication Date: 8/18/2024
Citation: Dassanayake, R.P., Menghwar, H., Bickel, K.A., Holthausen, D.J., Ma, H., Segunda, F., Rodriguez-Calzada, M., Medina, G.N., Attreed, S.E., Falkenberg, S.M., Kanipe, C.R., Sacco, R.E., De Los Santos, T.B., Casas, E. 2024. 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-28Ra). Frontiers in Immunology. 15. Article 1441908. https://doi.org/10.3389/fimmu.2024.1441908.
DOI: https://doi.org/10.3389/fimmu.2024.1441908

Interpretive Summary: Bovine viral diarrhea virus (BVDV) (noncytopathic) is the predominant biotype isolated from animals. BVDV is primarily a pathogen of cattle causing acute and persistent infections (PI), but it can also affect other domestic and wild ruminant species. Acute BVDV infections can lead to transient lymphocytopenia, the lymphocytes depletion in lymph nodes and thymus, and immunosuppression. The primary site of BVDV entry into cattle is oronasal route during acute infection and BVDV first replicates in nasal mucosa. Therefore, it is important to study the epithelial cells originated from nasal mucosa (turbinates) to find ways to reduce BVDV replication. In this study, we have tested anti-BVDV activity of a biotherapeutic agent (interferon lambda) in nasal epithelial cells. Our results suggested that reduced expression of one of the receptors (IFNLR1) as the reason for lack of anti-BVDV activity of interferon lambda in nasal epithelial cells.

Technical Abstract: Introduction: The antiviral activity of recombinant bovine interferon lambda 3 (bovIFN-'3) against bovine viral diarrhea virus (BVDV) has been demonstrated in vitro in Madin-Darby bovine kidney cells (MDBK) and in cattle in vivo. However, anti-BVDV activity of bovIFN-'3 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.Methods: Here we investigated the anti-BVDV activity of bovIFN-'3 in bovine turbinate-derived primary epithelial cells (BTu) using BVDV infection and staining, RT-qPCR, DNA and transcriptome sequencing, and transfection with plasmids containing bovIFN-'3 receptors (IL-28Ra and IL-10Rß).Results: Our results show that while BVDV was successfully cleared in MDBK cells treated with bovIFN-'3 and bovIFN-a, only the latter, bovIFN-a, cleared BVDV in BTu cells. Both cell types displayed intact type I and III IFN signaling pathways. Both cell types expressed similar levels of IL-10Rß subunit. DNA sequencing of amplified by PCR revealed intact transmembrane domain and lack of single nucleotide polymorphisms (SNPs) in the IL-28Ra subunit in BTu cells. However, RT-qPCR and transcriptomic analyses showed a lower expression of IL-28Ra transcripts in BTu cells as compared to MDBK cells. Interestingly, transfection of BTu cells with a plasmid encoding IL-28Ra subunit, but not IL-10Rß subunit, established the bovIFN-'3 sensitivity and induced anti-BVDV activity similar to the response in MDBK cells. Conclusion: Our results demonstrate that the sensitivity of cells to bovIFN-'3 depends not only on the quality but also of the quantity of the IL-28Ra subunit of the heterodimeric receptor. While no spliced variants or SNPs were identified in IL-28Ra of BTu cells, a reduced IL-28Ra transcript expression was detected in BTu as compared to MDBK cells. The establishment of bovIFN-'3 induced anti-BVDV activity in BTu cells transfected with IL-28Ra plasmid suggests that the levels of expression of this receptor subunit is crucial for the specific antiviral activity of type III IFN in these cells.