MHV-A59 fusion mutants are attenuated and display altered hepatotropism.
Mouse hepatitis virus strain A59 causes a persistent productive, but nonlytic, infection of cultured glial cells. We have mutants isolated from persistently infected glial cell cultures which have been shown to be fusion-defective due to a histidine to aspartic acid mutation (H716D) near the cleavage site of the peplomer protein, S. Here, we examine the pathogenicity of these mutants and show differences in hepatotropism and virulence compared to wild-type virus (WT). Two mutants chosen for detailed study, B11 and C12, were impaired in their abilities to cause hepatitis and/or replicate in the liver of susceptible mice. Furthermore, B11 and C12 display two separate hepatotropic phenotypes. The ability of B11 to replicate in the liver was dependent on infectious dose and route of inoculation, while C12 consistently displayed decreased hepatotropism regardless of dose and route of inoculation. However, B11 and C12 were shown to replicate in the CNS of infected animals similarly to WT. Like WT, the mutants produced meningoencephalitis during acute infection, with viral antigen exhibiting a similar distribution in the brain, and demyelination during chronic infection. Sequence analysis of wild-type, mutant, and revertant S proteins indicates that (1) a mutation in the N terminal subunit of S (S1), resulting in a glutamine to leucine amino acid substitution (Q159L), may affect hepatotropism and (2) a cleavage site mutation which determines fusogenicity is not responsible for altered hepatotropism. Furthermore, since B11, C12, and a nonattenuated fusion mutant (B12) have identical S protein sequences, there must be additional mutations outside of S which influence both virulence and hepatotropism.
Hingley
S T
ST
Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia 19104-6076.
Gombold
J L
JL
Lavi
E
E
Weiss
S R
SR
D000595:Amino Acid Sequence; D000818:Animals; D000956:Antigens, Viral / Q000235:genetics*; D001921:Brain / Q000382:microbiology / Q000473:pathology; D018352:Coronavirus Infections / Q000235:genetics*; D003711:Demyelinating Diseases / Q000382:microbiology; D008099:Liver / Q000382:microbiology* / Q000473:pathology; D008561:Membrane Fusion / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D006517:Murine hepatitis virus / Q000235:genetics / Q000472:pathogenicity*; D009154:Mutation; D009457:Neuroglia / Q000382:microbiology; D009928:Organ Specificity; D014764:Viral Proteins / Q000235:genetics*; D014774:Virulence
D000956:Antigens, Viral; D014764:Viral Proteins; C064484:spike protein S, Transmissible gastroenteritis virus