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Evaluation of replicative capacity and genetic stability of West Nile virus replicons using highly efficient packaging cell lines

This article is not included in your organization's subscription. However, you may be able to access this article under your organization's agreement with Elsevier.
Rafik Fayzulina, Frank Schollea, 1, Olga Petrakovab, Ilya Frolovb and Peter W. Masona, b, c, ,

aDepartment of Pathology, 3.206B Mary Moody Northen Pavilion, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX 77555-0436, USA

bDepartment of Microbiology and Immunology, UTMB, 301 University Boulevard, Galveston, TX 77555, USA

cSealy Center for Vaccine Development, UTMB, 301 University Boulevard, Galveston, TX 77555, USA

Received 6 February 2006; revised 23 February 2006; accepted 25 February 2006. Available online 2 May 2006.

Abstract
A stable cell system for high-efficiency packaging of West Nile virus (WNV) subgenomic replicons into virus-like particles (VLPs) was developed. VLPs could be propagated on these packaging cells and produced infectious foci similar to foci produced by WNV. Focus size correlated with the replicative capacity of WNV replicons, indicating that genome copy number, rather than amount of trans-complementing structural proteins, was rate-limiting in packaging of VLPs. Comparison of VLP production from replicon genomes encoding partial or complete C genes indicated that portions of C downstream of the cyclization sequence could improve genome replication or that cis expression of C could enhance packaging. Interestingly, a rapid loss of replicon-encoded reporter gene activity was detected within two serial passages of reporter gene-containing VLPs. The loss of reporter activity correlated with gene deletion and better VLP growth, indicating a powerful selection pressure for WNV genomes lacking reporter genes.

Keywords: Flavivirus; Replicon; Packaging; Virus-like particle; Noncytopathic replicon

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