title: An investigation of incorporation of cellular antigens into vaccinia virus particles.
abstract: Vaccinia virus (VV) infection produces several types of virus particle called intracellular mature virus (IMV), intracellular enveloped virus (IEV), cell-associated enveloped virus (CEV) and extracellular enveloped virus (EEV). Some cellular antigens are associated with EEV and these vary with the cell type used to grow the virus. To investigate if specific cell antigens are associated with VV particles, and to address the origin of membranes used to envelope IMV and IEV/CEV/EEV, we have studied whether cell antigens and foreign antigens expressed by recombinant VVs are incorporated into VV particles. Membrane proteins that are incorporated into the endoplasmic reticulum (ER), intermediate compartment (IC), cis/medial-Golgi, trans-Golgi network (TGN) or plasma membrane were not detected in purified IMV particles. In contrast, proteins present in the TGN or membrane compartments further downstream in the exocytic pathway co-purify with EEV particles when analysed by immunoblotting. Immunoelectron microscopy found only low levels of these proteins in IEV, CEV/EEV. The incorporation of foreign antigens into VV particles was not affected by loss of individual IEV or EEV-specific proteins or by redirection of B5R to the ER. These data suggest that (i) host cell antigens are excluded from the lipid envelope surrounding the IMV particle and (ii) membranes of the ER, IC and cis/medial-Golgi are not used to wrap IMV particles to form IEV. Lastly, the VV haemagglutinin was absent from one-third of IEV and CEV/EEV particles, whereas other EEV antigens were present in all these virions.
authors.affiliation: Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK1.
authors.forename: Oliver
authors.initials: O
authors.lastname: Krauss
authors.affiliation: Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK1.
authors.forename: Ruth
authors.initials: R
authors.lastname: Hollinshead
authors.affiliation: Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK1.
authors.forename: Michael
authors.initials: M
authors.lastname: Hollinshead
authors.affiliation: Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK1.
authors.forename: Geoffrey L
authors.initials: GL
authors.lastname: Smith
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D000944:Antigens, Differentiation, B-Lymphocyte / Q000378:metabolism; D018958:CD55 Antigens / Q000378:metabolism; D018957:CD59 Antigens / Q000378:metabolism; D002460:Cell Line; D002522:Chlorocebus aethiops; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D000077614:Golgi Matrix Proteins; D006367:HeLa Cells; D006389:Hemagglutinins, Viral; D015395:Histocompatibility Antigens Class I / Q000378:metabolism; D006801:Humans; D019012:Integrin beta1 / Q000378:metabolism; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D016253:Microscopy, Immunoelectron; D011990:Receptors, Transferrin; D060225:Tetraspanin 28; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D014616:Vaccinia virus / Q000235:genetics / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism; D015678:Viral Structural Proteins / Q000378:metabolism; D014771:Virion / Q000378:metabolism; D001613:beta 2-Microglobulin / Q000378:metabolism
chemical_list: C446940:A33R protein, Vaccinia virus; C081254:A34R protein, Vaccina virus; C400287:A36R protein, Vaccinia virus; C110361:A56R protein, Orthopoxvirus; D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; C497857:CD46 protein, human; D018958:CD55 Antigens; D018957:CD59 Antigens; C068126:CD71 antigen; C554798:CD81 protein, human; C081876:CKAP4 protein, human; D006023:Glycoproteins; D000077614:Golgi Matrix Proteins; D006389:Hemagglutinins, Viral; D015395:Histocompatibility Antigens Class I; D019012:Integrin beta1; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D011990:Receptors, Transferrin; D060225:Tetraspanin 28; D016159:Tumor Suppressor Protein p53; D014759:Viral Envelope Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; D001613:beta 2-Microglobulin; C087574:macrogolgin; C074512:42kDa protein, Vaccinia virus