Host-derived intracellular immunization against mouse hepatitis virus infection.
Mouse hepatitis virus (MHV) utilizes the murine biliary glycoprotein receptor (BgpA) for entry into susceptible cells. The Bgp1 gene was transfected into a murine DBT cell clone that expressed little if any BgpA receptor and resisted wild-type MHV infection. Clones which expressed low levels of receptor were efficient hosts for MHV-A59 replication. Clones which expressed 20- to 69-fold more BgpA receptor than controls were also susceptible to MHV-A59 infection, yet few infectious progeny virions were released. Pretreatment of clones with monoclonal antibody CC1, which binds to the N-terminus of BgpA, blocked MHV-A59 replication in DBT clones that expressed wild-type levels of receptor protein. In the overexpressing cell clones, however, CC1 pretreatment reversed the BgpA overexpression blockade and increased virus titers by 3-4 logs. BgpA overexpression inhibited the formation of infectious extracellular and intracellular virions, but levels of virus transcription were equivalent to those of controls. Ultrastructural studies revealed normal cell cytopathology in both the MHV-A59-infected controls and the overexpressing cell clones. Although equivalent numbers of virions were released compared with the control, peplomer spike glycoproteins were rarely evident in virions derived from the BgpA-overexpressing cell clones. Consonant with these findings, purified virions from BgpA-overexpressing cell clones demonstrated a 70% reduction in the amount of S glycoprotein, but not of N or M proteins. These data suggest that BgpA receptor overexpression establishes an intracellular trap which blocks MHV replication during the maturation and release of infectious progeny virions.
Chen
W
W
Department of Epidemiology, School of Public Health, University of North Carolina, Chapel Hill 27599, USA.
Madden
V J
VJ
Bagnell
C R
CR
Baric
R S
RS
D000818:Animals; D015703:Antigens, CD; D015815:Cell Adhesion Molecules; D002999:Clone Cells; D006023:Glycoproteins / Q000235:genetics* / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D006517:Murine hepatitis virus / Q000235:genetics / Q000502:physiology*; D011991:Receptors, Virus / Q000235:genetics* / Q000378:metabolism; D064370:Spike Glycoprotein, Coronavirus; D014158:Transcription, Genetic; D014162:Transfection; D014759:Viral Envelope Proteins / Q000378:metabolism; D014779:Virus Replication
D015703:Antigens, CD; C069847:CD66 antigens; D015815:Cell Adhesion Molecules; D006023:Glycoproteins; D008562:Membrane Glycoproteins; D011991:Receptors, Virus; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins