Chimaeric HIV-1 subtype C Gag molecules with large in-frame C-terminal polypeptide fusions form virus-like particles
| Autor: | Richard James Halsey, Sirika Pillay, Anna-Lise Williamson, Fiona Lesley Tanzer, Alisson Lynch, Ann E. Meyers, Edward P. Rybicki, Enid G. Shephard |
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| Rok vydání: | 2008 |
| Předmět: |
Cancer Research
Recombinant Fusion Proteins T-Lymphocytes viruses Blotting Western Human immunodeficiency virus (HIV) Gene Products gag Spodoptera Biology medicine.disease_cause complex mixtures Virus Mice Western blot Virology medicine Animals Humans Molecule A-DNA HIV vaccine Gene Cells Cultured Polyproteins AIDS Vaccines Mice Inbred BALB C medicine.diagnostic_test Immunogenicity Virion virus diseases Molecular biology Microscopy Electron Infectious Diseases HIV-1 Female Peptides Baculoviridae |
| Zdroj: | Virus Research. 133:259-268 |
| ISSN: | 0168-1702 |
| DOI: | 10.1016/j.virusres.2008.01.012 |
| Popis: | HIV-1 Pr55 Gag virus-like particles (VLPs) are strong immunogens with potential as candidate HIV vaccines. VLP immunogenicity can be broadened by making chimaeric Gag molecules: however, VLPs incorporating polypeptides longer than 200 aa fused in frame with Gag have not yet been reported. We constructed a range of gag-derived genes encoding in-frame C-terminal fusions of myristoylation-competent native Pr55Gag and p6-truncated Gag (Pr50Gag) to test the effects of polypeptide length and sequence on VLP formation and morphology, in an insect cell expression system. Fused sequences included a modified reverse transcriptase-Tat-Nef fusion polypeptide (RTTN, 778 aa), and truncated versions of RTTN ranging from 113 aa to 450 aa. Baculovirus-expressed chimaeric proteins were examined by western blot and electron microscopy. All chimaeras formed VLPs which could be purified by sucrose gradient centrifugation. VLP diameter increased with protein MW, from ∼100 nm for Pr55Gag to ∼250 nm for GagRTTN. The presence or absence of the Gag p6 region did not obviously affect VLP formation or appearance. GagRT chimaeric particles were successfully used in mice to boost T-cell responses to Gag and RT that were elicited by a DNA vaccine encoding a GagRTTN polypeptide, indicating the potential of such chimaeras to be used as candidate HIV vaccines. © 2008 Elsevier B.V. All rights reserved. |
| Databáze: | OpenAIRE |
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