title: Peptide presentation on primate erythroparvovirus 1 virus-like particles: In vitro assembly, stability and immunological properties.
abstract: Virus-like particles (VLPs) have demonstrated to be valuable scaffolds for the display of heterologous peptides for vaccine development and other specific interactions. VLPs of primate erythroparvovirus 1, generally referred as parvovirus B19 (B19V), have already been produced in-vivo and in-vitro from the recombinant VP2 protein of this virus. In this study, chimeric forms of B19V VP2 were constructed, and their ability to assemble into VLPs was evaluated. Chimeras were composed of the VP2 protein fused, at its N-terminus, with two peptides derived from the fusion glycoprotein (F) of the respiratory syncytial virus (RSV). The chimeric proteins self-assembled into VLPs morphologically similar to B19V virions. Stability of these VLPs was analyzed under denaturation conditions with guanidinium chloride (GdnHCl). Our results indicate that the presence of the heterologous fragments increased the stability of VLPs assembled by any of the VP2 chimeras. Specific proteolysis assays shown that a fraction of the N-termini of the chimeric proteins is located on the outer surface of the VLPs. Immunogenicity of VLPs against RSV was evaluated and the results indicate that the particles can elicit a humoral immune response, although these antibodies did not cross-react with RSV in ELISA tests. These results provide novel insights into the localization of the N-termini of B19V VP2 protein after in vitro assembly into VLPs, and point them to be attractive sites to display peptides or proteins without compromise the assembly or stability of VLPs.
authors: Department of Biochemistry, Faculty of Medicine, National Autonomous University of Mexico (UNAM), Mexico City 04510, Mexico.
authors: Areli
authors: A
authors: Del Carmen Morán-García
authors: Department of Microbiology and Parasitology, Faculty of Medicine, UNAM, Mexico City 04510, Mexico.
authors: Evelyn
authors: E
authors: Rivera-Toledo
authors: Department of Cell Biology, Faculty of Sciences, UNAM, Mexico City 04510, Mexico.
authors: Olga
authors: O
authors: Echeverría
authors: Department of Cell Biology, Faculty of Sciences, UNAM, Mexico City 04510, Mexico.
authors: Gerardo
authors: G
authors: Vázquez-Nin
authors: Department of Microbiology and Parasitology, Faculty of Medicine, UNAM, Mexico City 04510, Mexico.
authors: Beatriz
authors: B
authors: Gómez
authors: Department of Biochemistry, Faculty of Medicine, National Autonomous University of Mexico (UNAM), Mexico City 04510, Mexico. Electronic address: ismaelb@unam.mx.
authors: Ismael
authors: I
authors: Bustos-Jaimes
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D019791:Guanidine / Q000737:chemistry; D056724:Immunity, Humoral; D000071497:Immunogenicity, Vaccine / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D016732:Parvovirus B19, Human / Q000737:chemistry* / Q000276:immunology*; D019151:Peptide Library; D011489:Protein Denaturation; D055550:Protein Stability; D059748:Proteolysis; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D018113:Respiratory Syncytial Virus, Human / Q000737:chemistry / Q000276:immunology; D058425:Vaccines, Virus-Like Particle / Q000737:chemistry* / Q000276:immunology*; D014765:Viral Vaccines / Q000737:chemistry* / Q000276:immunology*; D014771:Virion / Q000737:chemistry / Q000235:genetics / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D058425:Vaccines, Virus-Like Particle; D014765:Viral Vaccines; C095548:capsid protein VP2, parvovirus B19; D019791:Guanidine
keywords: Parvovirus B19; Peptide display; Respiratory syncytial virus; Self-assembly; Virus-like particles