Characterization of Ebola Virus Entry by Using Pseudotyped Viruses: Identification of Receptor-Deficient Cell Lines

Abstract

Studies analyzing Ebola virus replication have been severely hampered by the extreme pathogenicity of this virus. To permit analysis of the host range and function of the Ebola virus glycoprotein (Ebo-GP), we have developed a system for pseudotyping these glycoproteins into murine leukemia virus (MLV). This pseudotyped virus, MLV(Ebola), can be readily concentrated to titers which exceed 5 × 106 infectious units/ml and is effectively neutralized by antibodies specific for Ebo-GP. Analysis of MLV(Ebola) infection revealed that the host range conferred by Ebo-GP is very broad, extending to cells of a variety of species. Notably, all lymphoid cell lines tested were completely resistant to infection; we speculate that this is due to the absence of a cellular receptor for Ebo-GP on B and T cells. The generation of high-titer MLV(Ebola) pseudotypes will be useful for the analysis of immune responses to Ebola virus infection, development of neutralizing antibodies, analysis of glycoprotein function, and isolation of the cellular receptor(s) for the Ebola virus.

Highly infectious SARS-CoV pseudotyped virus reveals the cell tropism and its correlation with receptor expression

“…Pseudotyped viruses with heterogenic glycoproteins incorporated into retroviral particles were proved to be a safe viral entry model, which only go through a single-cycle infection and acquired the host range of the parent viruses where the glycoproteins were derived [4][5][6], thus they could facilitate the research on viral entry mechanism, cell tropism, neutralization antibody analysis, and receptor identification [7][8][9][10].…”

Characterization of retrovirus-based reporter viruses pseudotyped with the precursor membrane and envelope glycoproteins of four serotypes of dengue viruses

“…Data are means of duplicate wells of one representative experiment and the error bars represent the standard errors of the means. Matsuura et al, 2001;Moore et al, 2004;Nie et al, 2004;Simmons et al, 2004;Wool-Lewis and Bates, 1998). In this study, we have successfully established a retrovirus-based pseudotype reporter virus for four serotypes of dengue viruses.…”

“…Like other flaviviruses, dengue virus is believed to enter cells through receptor-mediated endocytosis, a process sensitive to low pH environment in the endosome (Guirakhoo et al, 1993;Lindenbach and Rice, 2001;Mellman et al, 1986;Mukhopadhyay et al, 2005;. To investigate entry of the pseudotype reporter virus, Huh7 cells were treated with increasing concentrations of two lysosomotropic agents, Bafilomycin A and ammonium chloride, and infected with D2(HIVluc) and VSV(HIVluc), which has been shown to enter cells through a pH-dependent route Nie et al, 2004;Simmons et al, 2004;Wool-Lewis and Bates, 1998). As shown in Fig.…”

“…Pseudotype reporter viruses that contain the E protein of one virus and the core and genome with reporter gene of another virus have been shown to be a convenient molecular tool to study the functions of E protein, such as tropism, mechanism of entry, sensitivity to neutralization, enhancement and entry inhibitors (Bartosch et al, 2003;Buonocore et al, 2002;Chan et al, 2001;Cronin et al, 2005;Deng et al, 1997;Fukushi et al, 2005;Giroglou et al, 2004;Hanika et al, 2005;Hsu et al, 2003;Ma et al, 1999;Matsuura et al, 2001;Moore et al, 2004;Nie et al, 2004;Simmons et al, 2004;Wool-Lewis and Bates, 1998). Retrovirus-based reporter viruses pseudotyped with the E proteins of different families of RNA viruses, including simian immunodeficiency virus, hepatitis C virus (HCV), Ebola virus, La Crosse virus, Hantaan virus, and SARS-CoV, have been successfully established (Bartosch et al, 2003;Chan et al, 2001;Deng et al, 1997;Giroglou et al, 2004;Hsu et al, 2003;Ma et al, 1999;Moore et al, 2004;Nie et al, 2004;Simmons et al, 2004;Wool-Lewis and Bates, 1998). In this study, we developed a lentivirus-based pseudotype reporter virus for dengue virus, which contains the core and defective genome of human immunodeficiency virus type 1 (HIV-1), the luciferase reporter gene, and the PrM/ E proteins of one of the four serotypes of dengue viruses.…”

Optimal control application to an Ebola model

“…It belongs to the genus Ebolavirus, within the family of Filoviridae. Its envelope glycoprotein facilitates the entry of the virus to living cells [7,8] . Till date five strains of the virus are known: Zaire, Sudan, Tai Forest, Bundibugyo, and Reston with Zaire stain being the most virulent with up to 90% fatalities [9].…”