Entry of bovine viral diarrhea virus into ovine cells occurs through clathrin-dependent endocytosis and low pH-dependent fusion (original) (raw)

Abstract

Although mechanisms of bovine viral diarrhea virus (BVDV) entry into bovine cells have been elucidated, little is known concerning pestivirus entry and receptor usage in ovine cells. In this study, we determined the entry mechanisms of BVDV-1 and BVDV-2 in sheep fetal thymus cells. Both BVDV-1 and BVDV-2 infections were inhibited completely by chlorpromazine, β-methyl cyclodextrin, sucrose, bafilomycin A1, chloroquine, and ammonium chloride. Simultaneous presence of reducing agent and low pH resulted in marked loss of BVDV infectivity. Moreover, BVDV was unable to fuse with ovine cell membrane by the presence of reducing agent or low pH alone, while combination of both led to fusion at low efficiency. Furthermore, sheep fetal thymus cells acutely infected with BVDV-1 or BVDV-2 were found protected from heterologous BVDV infection. Taken together, our results showed for the first time that entry of both BVDV-1 and BVDV-2 into ovine cells occurred through clathrin-dependent endocytosis, endosomal acidification, and low pH-dependent fusion following an activation step, besides suggesting the involvement of a common ovine cellular receptor during attachment and entry.

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Acknowledgments

We thank Dr. Roland Riebe, Collection of Cell Lines in Veterinary Medicine, Friedrich-Loeffler Institute, Island of Riems, Germany for supplying the SFT-R cell line. This work was supported by a grant from Department of Biotechnology, Govt. of India, New Delhi.

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Authors and Affiliations

1. High Security Animal Disease Laboratory, Indian Veterinary Research Institute, Anand Nagar, Bhopal, 462021, Madhya Pradesh, India
   Basavaraj Shrishail Mathapati, Niranjan Mishra, Katherukamem Rajukumar, Ram Kumar Nema, Sthita Pragnya Behera & Shiv Chandra Dubey

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1. Basavaraj Shrishail Mathapati
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2. Niranjan Mishra
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3. Katherukamem Rajukumar
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4. Ram Kumar Nema
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5. Sthita Pragnya Behera
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6. Shiv Chandra Dubey
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Correspondence toNiranjan Mishra.

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Editor: J. Denry Sato

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Mathapati, B.S., Mishra, N., Rajukumar, K. et al. Entry of bovine viral diarrhea virus into ovine cells occurs through clathrin-dependent endocytosis and low pH-dependent fusion.In Vitro Cell.Dev.Biol.-Animal 46, 403–407 (2010). https://doi.org/10.1007/s11626-009-9263-9

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• Received: 01 September 2009
• Accepted: 20 October 2009
• Published: 08 December 2009
• Issue Date: May 2010
• DOI: https://doi.org/10.1007/s11626-009-9263-9

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