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Mature HIV-like Particles Produced from Single Semliki Forest Virus-Derived Expression Vector  

KIM EUN (Laboratory of Infectious Disease, College of Veterinary Medicine, Chungnam National University)
POO HAR-YOUNG (Proteome Research Lab., Korea Research Institute of Bioscience and Biotechnology)
SUNG MOON-HEE (Department of Bio & Nanochemisty, College of Natural Sciences, Kookmin University)
KIM CHUL-JOONG (Laboratory of Infectious Disease, College of Veterinary Medicine, Chungnam National University)
Publication Information
Journal of Microbiology and Biotechnology / v.15, no.6, 2005 , pp. 1229-1239 More about this Journal
Abstract
Human immunodeficiency virus-like particles (HIVVLPs) with native conformations similar to that of the wild-type virion could be valid candidates for vaccine development. To this end, we used a Semliki Forest Virus (SFV) expression system to produce HIV- VLPs containing high quantities of native envelope proteins. Here, we described a single SFV replicon containing the HIV gagpol and env genes under the control of separate subgenomic promoters. Mature VLPs incorporating the Gag and Env proteins were detected in the supernatant of replicon-expressing cells by Western blot analysis. The HIV-VLPs showed the expected molecular density (1.14-1.18 g/ml) on a $20-60\%$ sucrose gradient; the particles were 100-120 nm in diameter and Env proteins were observed on their surfaces by immunogold electron microscopy. RT-PCR analysis of VLP-associated RNAs in mature HIV-VLPs revealed two SF V-derived RNA species (full-length and subgenomic). Immunization studies in Balb/c mice showed that these HIV-VLPs were capable of inducing both HIV-specific antibodies and cell-mediated immune responses. Taken together, our results indicate that the SFV replicon system is useful for the production of HIV-VLPs, which may be valuable candidates for an HIV vaccine.
Keywords
HIV-1; Semliki Forest Virus vector; virus-like particle; vaccine;
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