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Journal of Virology, November 2004, p. 11605-11614, Vol. 78, No. 21
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.21.11605-11614.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

trans-Packaged West Nile Virus-Like Particles: Infectious Properties In Vitro and in Infected Mosquito Vectors

Frank Scholle,1,2* Yvette A. Girard,1 Qizu Zhao,1,{dagger} Stephen Higgs,1 and Peter W. Mason1,3

Department of Pathology,1 Department of Microbiology and Immunology,2 Sealy Center for Vaccine Development, University of Texas Medical Branch, Galveston, Texas3

Received 5 March 2004/ Accepted 27 June 2004

A trans-packaging system for West Nile virus (WNV) subgenomic replicon RNAs (repRNAs), deleted for the structural coding region, was developed. WNV repRNAs were efficiently encapsidated by the WNV C/prM/E structural proteins expressed in trans from replication-competent, noncytopathic Sindbis virus-derived RNAs. Infectious virus-like particles (VLPs) were produced in titers of up to 109 infectious units/ml. WNV VLPs established a single round of infection in a variety of different cell lines without production of progeny virions. The infectious properties of WNV and VLPs were indistinguishable when efficiencies of infection of a number of different cell lines and inhibition of infection by neutralizing antibodies were determined. To investigate the usefulness of VLPs to address biological questions in vivo, Culex pipiens quinquefasciatus mosquitoes were orally and parenterally infected with VLPs, and dissected tissues were analyzed for WNV antigen expression. Antigen-positive cells in midguts of orally infected mosquitoes were detected as early as 2 days postinfection and as late as 8 days. Intrathoracic inoculation of VLPs into mosquitoes demonstrated a dose-dependent pattern of infection of secondary tissues and identified fat body, salivary glands, tracheal cells, and midgut muscle as susceptible WNV VLP infection targets. These results demonstrate that VLPs can serve as a valuable tool for the investigation of tissue tropism during the early stages of infection, where virus spread and the need for biosafety level 3 containment complicate the use of wild-type virus.

* Corresponding author. Mailing address: Department of Pathology, 3.218 Mary Moody Northen, University of Texas Medical Branch, 301 University Blvd., Galveston, TX 77555-0436. Phone: (409) 747-8158. Fax: (409) 747-8150. E-mail: frscholl{at}utmb.edu.

{dagger} Present address: Institute of Biophysics, Chinese Academy of Sciences, Beijing, China 100101.

Journal of Virology, November 2004, p. 11605-11614, Vol. 78, No. 21
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.21.11605-11614.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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