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The binding of dengue type 2 (DEN-2) virus to mammalian (LLC-MK2 and Vero) and mosquito (C6/36 and AP61) cell surfaces was investigated by a virus-binding assay using purified 3H-labeled DEN-2 virus. The DEN-2 virus binding to all four cell types was specific and saturable, indicating the presence of a single class of receptors (ranging from 3.7 x 103 to 3.5 x 104 receptors/cell) with a high affinity for DEN-2 virus (Kd ranging from 98 to 171 pM). Treatment of cell surfaces with certain glycosidases significantly reduced virus binding to mammalian cell lines, but not to the insect cell lines examined. Furthermore, heparin was found to compete with mammalian cell receptors for binding to DEN-2 virus and to inhibit viral infection of mammalian cells, but heparin had no effect on viral binding to or infection of insect cells. These results confirm previous reports suggesting that DEN-2 virus receptors on mammalian cell lines are different from those on insect cell lines.
Received August 29, 2003. Accepted for publication September 25, 2003.
Acknowledgments: We thank Dr. Ananda Nisalak and Dr. Khin Saw Aye Myint for their valuable suggestions, Somsak Imlarp for cell culture assistance, and Dr. Mammen P. Mammen for his review of the manuscript.
Financial support: This work was supported by the United States Army Medical Research and Materiel Command, (Fort Detrick, Frederick, MD).
Disclaimer: The opinions or assertions contained herein are the private views of the authors and are not to be construed as reflecting the official views of the United States Army or the Department of Defense.
Authors’ addresses: Butsaya K. Thaisomboonsuk, Department of Virology, United States Army Medical Component, Armed Forces Research Institute of Medical Sciences, APO AP 96546 (from the United States) or 315/6 Rajvithi Road, Bangkok 10400, Thailand (from outside the United States), Telephone: 66-2-644-5644, Fax: 66-2-644-4760, E-mail: ButsayaT{at}afrims.org Edward T. Clayson, Project Management Office, Chemical Biological Medical Systems, 64 Thomas Johnson Drive, Frederick, Maryland 21702, Telephone: 301-619-8402, Fax: 301-619-8025, E-mail: Edward.Clayson{at}det.amedd.army.mil. Somsak Pantuwatana, Department of Microbiology, Mahidol University, Bangkok 10400, Thailand, E-mail: pantuwan{at}bucc4.buu.ac.th. David W. Vaughn, Military Infectious Diseases Research Program, Medical Research and Materiel Command, 504 Scott Street, Fort Detrick, Frederick, MD 21702-5012, E-mail: David. Vaughn{at}amedd.army.mil. Timothy P. Endy, Walter Reed Army Institute of Research, Rm 3S28, 503 Robert Grant Ave., Silver Spring, MD 20910-7500, E-mail: Timothy.Endy{at}na.amedd.army.mil.
Reprint requests: Butsaya K. Thaisomboonsuk, Department of Virology, United States Army Medical Component, Armed Forces Research Institute of Medical Sciences, APO AP 96546 (from the United States) or 315/6 Rajvithi Road, Bangkok 10400, Thailand (from outside the United States).
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