Isolation, characterization, evaluation of pathogenicity, and immunomodulation through interferon production of duck adenovirus type-3 (DAdV-3)

Abstract

Duck adenovirus type-3 (DAdV-3) is a poorly characterized duck virus. A comprehensive analysis of the DAdV-3 pathogenicity and host immune response could be a valuable addition. Herein, DAdV-3 was isolated from Muscovy duck and virus-specific genes were confirmed by polymerase chain reaction (PCR). The obtained gene fragments were sequenced and compared with the reference sequence. Results confirmed that the clinically isolated virus was DAdV-3, named as HF-AN-2020. To evaluate DAdV-3 host immune response, the expression levels of MDA5, STING, IRF7, MAVS, and NF-κB, and inflammatory cytokines (IFN-β, IFN-γ, and IL-1β) were determined by quantitative reverse transcriptase PCR (qRT-PCR). The expression levels of IFN-β and IFN-γ were 32.6- and 28.6-fold, respectively, higher (P < 0.01) than the control group. It was found that the upregulation of STING and NF-κB pathways was directly involved in the regulation of inflammatory cytokines (IFN-β, IFN-γ, and IL-1β). Furthermore, the gene regulation pathways consecutively upregulated the expression levels of MDA5, STING, IRF7, MAVS, and NF-κB up to 31.6, 10.5, 31.4, 2.2, and 2.6-fold, respectively, higher (P < 0.01) than the control group. The TCID50 of DAdV-3 for Muscovy duck and chicken was 10-3.24/0.1 mL with 0% mortality, indicating low pathogenicity in both Muscovy ducks and chickens, but DAdV-3 can induce higher expression of interferons. Genome analysis showed mutations in 4 amino acids located in ORF19B (Ser to Thr), ORF66 (Leu to Phe, Ile to Leu), and ORF67 (Gly to stop codon). This study provides essential and basic information for further research on the mechanism of the cellular immune responses against adenoviruses.