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Title: Characterization of dengue virus 2 growth in megakaryocyte–erythrocyte progenitor cells

Abstract

Megakaryocyte–erythrocyte progenitor (MEP) cells are potential in vivo targets of dengue virus (DENV); the virus has been found associated with megakaryocytes ex vivo and platelets during DENV-induced thrombocytopenia. We report here that DENV serotype 2 (DENV2) propagates well in human nondifferentiated MEP cell lines (Meg01 and K562). In comparison to virus propagated in Vero cells, viruses from MEP cell lines had similar structure and buoyant density. However, differences in MEP-DENV2 stability and composition were suggested by distinct protein patterns in western blot analysis. Also, antibody neutralization of envelope domain I/II on MEP-DENV2 was reduced relative to that on Vero-DENV2. Infectious DENV2 was produced at comparable kinetics and magnitude in MEP and Vero cells. However, fewer virion structures appeared in electron micrographs of MEP cells. We propose that DENV2 infects and produces virus efficiently in megakaryocytes and that megakaryocyte impairment might contribute to dengue disease pathogenesis. - Highlights: • DenV replicates efficiently in undifferentiated megakaryocyte–erythrocyte progenitors. • MEP produced DenV differs in protein content from Vero produced DenV. • MEP produced DenV may be more difficult to neutralize relative to Vero DenV.

Authors:
 [1]; ;  [2];  [3];  [2];  [4];  [1];  [5]
  1. Division of Microbiology and Immunology, Yerkes National Primate Research Center, Emory University, Atlanta, GA (United States)
  2. Center for AIDS Research, Department of Pediatrics, Emory University School of Medicine and Veterans Affairs Medical Center, Atlanta, GA (United States)
  3. Departments of Pediatrics, Pathology, Microbiology, and Immunology, Vanderbilt University, Nashville, TN (United States)
  4. Department of Microbiology and Immunology, College of Medicine, National Cheng Kung University, Tainan, Taiwan (China)
  5. (United States)
Publication Date:
OSTI Identifier:
22581693
Resource Type:
Journal Article
Resource Relation:
Journal Name: Virology; Journal Volume: 493; Other Information: Copyright (c) 2016 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ANTIBODIES; BONE MARROW CELLS; ELECTRON MICROSCOPY; ERYTHROCYTES; IN VIVO; PATHOGENESIS; PROTEINS; VIRAL DISEASES; VIRUSES

Citation Formats

Clark, Kristina B., Hsiao, Hui-Mien, Bassit, Leda, Crowe, James E., Schinazi, Raymond F., Perng, Guey Chuen, Villinger, Francois, and New Iberia Research Center, University of Louisiana at Lafayette, New Iberia, LA. Characterization of dengue virus 2 growth in megakaryocyte–erythrocyte progenitor cells. United States: N. p., 2016. Web. doi:10.1016/J.VIROL.2016.03.024.
Clark, Kristina B., Hsiao, Hui-Mien, Bassit, Leda, Crowe, James E., Schinazi, Raymond F., Perng, Guey Chuen, Villinger, Francois, & New Iberia Research Center, University of Louisiana at Lafayette, New Iberia, LA. Characterization of dengue virus 2 growth in megakaryocyte–erythrocyte progenitor cells. United States. doi:10.1016/J.VIROL.2016.03.024.
Clark, Kristina B., Hsiao, Hui-Mien, Bassit, Leda, Crowe, James E., Schinazi, Raymond F., Perng, Guey Chuen, Villinger, Francois, and New Iberia Research Center, University of Louisiana at Lafayette, New Iberia, LA. 2016. "Characterization of dengue virus 2 growth in megakaryocyte–erythrocyte progenitor cells". United States. doi:10.1016/J.VIROL.2016.03.024.
@article{osti_22581693,
title = {Characterization of dengue virus 2 growth in megakaryocyte–erythrocyte progenitor cells},
author = {Clark, Kristina B. and Hsiao, Hui-Mien and Bassit, Leda and Crowe, James E. and Schinazi, Raymond F. and Perng, Guey Chuen and Villinger, Francois and New Iberia Research Center, University of Louisiana at Lafayette, New Iberia, LA},
abstractNote = {Megakaryocyte–erythrocyte progenitor (MEP) cells are potential in vivo targets of dengue virus (DENV); the virus has been found associated with megakaryocytes ex vivo and platelets during DENV-induced thrombocytopenia. We report here that DENV serotype 2 (DENV2) propagates well in human nondifferentiated MEP cell lines (Meg01 and K562). In comparison to virus propagated in Vero cells, viruses from MEP cell lines had similar structure and buoyant density. However, differences in MEP-DENV2 stability and composition were suggested by distinct protein patterns in western blot analysis. Also, antibody neutralization of envelope domain I/II on MEP-DENV2 was reduced relative to that on Vero-DENV2. Infectious DENV2 was produced at comparable kinetics and magnitude in MEP and Vero cells. However, fewer virion structures appeared in electron micrographs of MEP cells. We propose that DENV2 infects and produces virus efficiently in megakaryocytes and that megakaryocyte impairment might contribute to dengue disease pathogenesis. - Highlights: • DenV replicates efficiently in undifferentiated megakaryocyte–erythrocyte progenitors. • MEP produced DenV differs in protein content from Vero produced DenV. • MEP produced DenV may be more difficult to neutralize relative to Vero DenV.},
doi = {10.1016/J.VIROL.2016.03.024},
journal = {Virology},
number = ,
volume = 493,
place = {United States},
year = 2016,
month = 6
}
  • Dengue virus causes {approx}50-100 million infections per year and thus is considered one of the most aggressive arthropod-borne human pathogen worldwide. During its replication, dengue virus induces dramatic alterations in the intracellular membranes of infected cells. This phenomenon is observed both in human and vector-derived cells. Using high-resolution mass spectrometry of mosquito cells, we show that this membrane remodeling is directly linked to a unique lipid repertoire induced by dengue virus infection. Specifically, 15% of the metabolites detected were significantly different between DENV infected and uninfected cells while 85% of the metabolites detected were significantly different in isolated replication complexmore » membranes. Furthermore, we demonstrate that intracellular lipid redistribution induced by the inhibition of fatty acid synthase, the rate-limiting enzyme in lipid biosynthesis, is sufficient for cell survival but is inhibitory to dengue virus replication. Lipids that have the capacity to destabilize and change the curvature of membranes as well as lipids that change the permeability of membranes are enriched in dengue virus infected cells. Several sphingolipids and other bioactive signaling molecules that are involved in controlling membrane fusion, fission, and trafficking as well as molecules that influence cytoskeletal reorganization are also up regulated during dengue infection. These observations shed light on the emerging role of lipids in shaping the membrane and protein environments during viral infections and suggest membrane-organizing principles that may influence virus-induced intracellular membrane architecture.« less
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