Multiscale model of defective interfering particle replication for influenza A virus infection in animal cell culture

• Main Authors: Hein, M.D (Author), Kupke, S.Y (Author), Pelz, L. (Author), Reichl, U. (Author), Rüdiger, D. (Author)
• Format: Article
• Language: English
• Published: Public Library of Science 2021
• Subjects: animal; animal cell; animal cell culture; Animals; Antiviral Agents; antiviral therapy; antivirus agent; apoptosis; Article; biological model; cell culture technique; Cell Culture Techniques; centrifugation; chemistry; coinfection; controlled study; defective virus; Defective Viruses; dog; Dogs; dynamics; flow cytometry; gene deletion; genetics; glutamine; influenza A; Influenza A virus; Madin Darby Canine Kidney Cells; mathematical model; MDCK cell line; Models, Biological; nonhuman; Orthomyxoviridae Infections; orthomyxovirus infection; pathogenicity; physiology; plaque forming unit; prediction; real time reverse transcription polymerase chain reaction; ribonucleoprotein; RNA directed RNA polymerase; RNA, Viral; sialidase; simulation; viral protein; virology; virus infection; virus particle; virus release; virus replication; Virus Replication; virus RNA; virus transmission
• Online Access: View Fulltext in Publisher

 Cell culture-derived defective interfering particles (DIPs) are considered for antiviral therapy due to their ability to inhibit influenza A virus (IAV) production. DIPs contain a large internal deletion in one of their eight viral RNAs (vRNAs) rendering them replication-incompetent. However, they can propagate alongside their homologous standard virus (STV) during infection in a competition for cellular and viral resources. So far, experimental and modeling studies for IAV have focused on either the intracellular or the cell population level when investigating the interaction of STVs and DIPs. To examine these levels simultaneously, we conducted a series of experiments using highly different multiplicities of infections for STVs and DIPs to characterize virus replication in Madin-Darby Canine Kidney suspension cells. At several time points post infection, we quantified virus titers, viable cell concentration, virus-induced apoptosis using imaging flow cytometry, and intracellular levels of vRNA and viral mRNA using real-time reverse transcription qPCR. Based on the obtained data, we developed a mathematical multiscale model of STV and DIP co-infection that describes dynamics closely for all scenarios with a single set of parameters. We show that applying high DIP concentrations can shut down STV propagation completely and prevent virus-induced apoptosis. Interestingly, the three observed viral mRNAs (full-length segment 1 and 5, defective interfering segment 1) accumulated to vastly different levels suggesting the interplay between an internal regulation mechanism and a growth advantage for shorter viral RNAs. Furthermore, model simulations predict that the concentration of DIPs should be at least 10000 times higher than that of STVs to prevent the spread of IAV. Ultimately, the model presented here supports a comprehensive understanding of the interactions between STVs and DIPs during co-infection providing an ideal platform for the prediction and optimization of vaccine manufacturing as well as DIP production for therapeutic use. Copyright: © 2021 Rüdiger et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.