Molecular Cloning and Expression of Major Structural Protein VP1 of the Human Polyomavirus JC Virus: Formation of Virus-Like Particles Useful for Immunological and Therapeutic Studies

Abstract

The major structural viral protein, VP1, of the human polyomavirus JC virus (JCV), the causative agent of progressive multifocal leukoencephalopathy (PML), was expressed by using recombinant baculoviruses. Recombinant VP1 formed virus-like particles (VLP) with the typical morphology of empty JCV capsids. Purified VP1 VLP bind to SVG, B, and T cells, as well as to monkey kidney cells. After binding, VP1 VLP were also internalized with high efficiency and transported to the nucleus. Immunization studies revealed these particles as highly immunogenic when administered with adjuvant, while immunization without adjuvant induced no immune response. VP1 VLP hyperimmune serum inhibits binding to SVG cells and neutralizes natural JCV. Furthermore, the potential of VP1 VLP as an efficient transporter system for gene therapy was demonstrated. Exogenous DNA could be efficiently packaged into VP1 VLP, and the packaged DNA was transferred into COS-7 cells as shown by the expression of a marker gene. Thus, VP1 VLP are useful for PML vaccine development and represent a potential new transporter system for human gene therapy.

A replication-incompetent Rift Valley fever vaccine: Chimeric virus-like particles protect mice and rats against lethal challenge

“…Several VLP-based vaccines are in human clinical trials or are FDAapproved, including those for hepatitis B virus (Andre and Safary, 1987;McAleer et al, 1984;Sitrin et al, 1993), trivalent influenza H1N1, H3N2 and B vaccine ("FluBlok") (Cox et al, 2008;Treanor et al, 2007), H5N1 "bird flu" (Perrone et al, 2009), human papillomavirus Giannini et al, 2006;Harper et al, 2004;Harro et al, 2001;Joura et al, 2007), human immunodeficiency virus (Young et al, 2006) and Norwalk virus (Tacket et al, 2003). Other VLP-based vaccines with very promising pre-clinical results include VLPs for the severe acute respiratory syndrome (SARS) coronavirus (Lokugamage et al, 2008), human polyomavirus (Goldmann et al, 1999), rotavirus (Ciarlet et al, 1998;El-Attar et al, 2009;Jiang et al, 1999), and Ebola and Marburg viruses (Swenson et al, 2008;Swenson et al, 2005;Warfield et al, 2003;Warfield et al, 2005b;Warfield et al, 2007). Additionally, VLP vaccines also have important agricultural applications, including promising vaccines for livestock diseases including bluetongue (Noad and Roy, 2003;Roy, 2000;Roy et al, 2009;Roy and Noad, 2008) and foot-and-mouth (Li et al, 2008;Remond et al, 2009).…”

Virus-like particle-based vaccines for animal viral infections

“…9,10,23,24 However, this statement should be tempered by the fact that some VLP-based candidate vaccines require formulation with potent adjuvants in order to induce efficient immune responses, indicating that the relative ability of diverse VLP types to induce the different branches of the immune response is influenced by a number of factors that are VLP-specific. 25,26 Overall, VLPs have been shown to stimulate strong B-cell-mediated immune responses and can be highly effective at stimulating CD4 + T cell proliferative responses and cytotoxic T lymphocyte (CTL) responses, [27][28][29][30] the fundamental goal for any vaccine. The immune system has multiple mechanisms to robustly respond to virus particles, 10,31 which may be exploited by VLP-based vaccines.…”

Virus-like particles: The new frontier of vaccines for animal viral infections

“…The fact that VLPs present overall suitable characteristics for their uptake by DCs and subsequent processing and presentation by MHC-II and MHC-I (cross-presentation) pathways, led to describe VLPs as "self-adjuvanting" immunogen delivery systems (Grgacic and Anderson, 2006;Liu et al, 2000;Ludwig and Wagner, 2007;Yan et al, 2004). However, this statement should be tempered by the fact that some VLP-based candidate vaccines require formulation with potent adjuvants in order to induce efficient immune responses ( Table 2), indicating that the relative ability of diverse VLP types to induce the different branches of the immune response is influenced by a number of factors that are VLP-specific (Gedvilaite et al, 2006;Goldmann et al, 1999).…”