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Open AccessArticle

Real Time Analysis of Bovine Viral Diarrhea Virus (BVDV) Infection and Its Dependence on Bovine CD46

Institute of Virology, Vetmeduni Vienna, 1210 Vienna, Austria
VetCore Facility for Research, Vetmeduni Vienna, 1210 Vienna, Austria
Institute of Virology, Faculty of Veterinary Medicine, Justus-Liebig University, 35392 Gießen, Germany
Department of Infectious Diseases, Virology, University of Heidelberg, 69120 Heidelberg, Germany
German Center for Infection Research, 69120 Heidelberg, Germany
Author to whom correspondence should be addressed.
Viruses 2020, 12(1), 116;
Received: 16 December 2019 / Revised: 10 January 2020 / Accepted: 14 January 2020 / Published: 17 January 2020
(This article belongs to the Special Issue Bovine Viral Diarrhea Virus and Related Pestiviruses)
Virus attachment and entry is a complex interplay of viral and cellular interaction partners. Employing bovine viral diarrhea virus (BVDV) encoding an mCherry-E2 fusion protein (BVDVE2-mCherry), being the first genetically labelled member of the family Flaviviridae applicable for the analysis of virus particles, the early events of infection—attachment, particle surface transport, and endocytosis—were monitored to better understand the mechanisms underlying virus entry and their dependence on the virus receptor, bovine CD46. The analysis of 801 tracks on the surface of SK6 cells inducibly expressing fluorophore labelled bovine CD46 (CD46fluo) demonstrated the presence of directed, diffusive, and confined motion. 26 entry events could be identified, with the majority being associated with a CD46fluo positive structure during endocytosis and occurring more than 20 min after virus addition. Deletion of the CD46fluo E2 binding domain (CD46fluo∆E2bind) did not affect the types of motions observed on the cell surface but resulted in a decreased number of observable entry events (2 out of 1081 tracks). Mean squared displacement analysis revealed a significantly increased velocity of particle transport for directed motions on CD46fluo∆E2bind expressing cells in comparison to CD46fluo. These results indicate that the presence of bovine CD46 is only affecting the speed of directed transport, but otherwise not influencing BVDV cell surface motility. Instead, bovine CD46 seems to be an important factor during uptake, suggesting the presence of additional cellular proteins interacting with the virus which are able to support its transport on the virus surface. View Full-Text
Keywords: Pestivirus; BVDV; CD46; life cell imaging; attachment; surface transport Pestivirus; BVDV; CD46; life cell imaging; attachment; surface transport
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Riedel, C.; Chen, H.-W.; Reichart, U.; Lamp, B.; Laketa, V.; Rümenapf, T. Real Time Analysis of Bovine Viral Diarrhea Virus (BVDV) Infection and Its Dependence on Bovine CD46. Viruses 2020, 12, 116.

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