Mouse hepatitis virus type 4 (JHM strains). induced fatal central nervous system disease. I. genetic control and murine neuron as the susceptible site of disease.

Abstract

Mouse hepatitis virus (JHM strain) type 4 induces acute encephalitis followed by death in many strains of laboratory mice. Immunohistochemical study in vivo and analysis of mouse neuronal cells in vitro both indicate that the target cells in this infection is the neuron. Further, examination of several inbred mouse strains and neuronal cells from them shows that disease expression is controlled by a single autosomal gene action at the level of the neuronal cell. Susceptibility is dominant but not H-2 linked. However, cultured neuronal cells and macrophages from SJL/J mice, which are resistant to this infection, fail to make significant amounts of infectious virus after an appropriate viral inoculation. Apparently the defect is not at the level of the virus-cell receptor, because these cells, in part, express viral antigens.

Host Genetic Control of Mouse Hepatitis Virus Type 4 (JHM Strain) Replication. I. Restriction of Virus Amplification and Spread in Macrophages from Resistant Mice

“…Cultured cells on glass coverslips were fixed with 4~ paraformaldehyde and 0.5% glutaraldehyde in 0.1 M-phosphate buffer pH 7-3, washed in phosphate-buffered saline (PBS), and incubated with either polyvalent mouse antibody to mouse hepatitis virus (provided by the Resource Branch of the National Cancer Institute, Bethesda, Md., U.S.A.) or monoclonal antibodies to MHV-4 (Collins et al, 1982) for 30 min. The coverslips were washed three times in PBS and incubated with the F(ab)~ fragment of rabbit antibody to mouse IgG, conjugated with fluorescein isothiocyanate or peroxidase (Cappel Laboratories, Cochranville, Pa., U.S.A.) for 30 min, as reported by Knobler et al (1981). All reactions took place at room temperature.…”

“…The peritoneal cavity was lavaged with 5 to 6 ml of sterile Eagle's MEM which contained 20 units/ml heparin, 1 glutamine, 100 ktg/ml penicillin and streptomycin. Cells were plated to provide 1 × 106 adherent cells/dish as reported previously (Brautigam et al, 1979;Knobler et al, 1981), and identified as macrophages by their ability to ingest zymosan particles or sheep erythrocytes previously reacted with antibody and C5-deficient mouse serum, and by their morphology (van Furth et al, 1978). Homogeneity was usually > 95~ with a range of 95 to 99~.…”

“…The handling of virus, culture and plaque assay conditions have been reported elsewhere (Haspel et al, 1978;Knobler et al, 1981). Vesicular stomatitis virus (VSV), Indiana strain, was titrated by plaque assay on L-929 cells (Casali et al, 1981).…”

“…Resistance to MHV-4-induced fatal central nervous system disease is inherited as a single autosomal recessive trait, independent of the major histocompatibility complex, expressed at the level of the neuronal cell (Knobler et al, 1981). Resistance is mirrored by peritoneal macrophages elicited from SJL/J mice (Stohlman & Frelinger, 1981;Knobler et al, 1981).…”

“…Mouse hepatitis virus type 4 (MHV-4, JHM strain), a member of the coronavirus family, is attracting increasing interest as an agent of both virus-induced encephalomyelitis and virusinduced demyelination (Bailey et al, 1949;Waksman & Adams, 1962;Lampert et al, 1973 ;Weiner, 1973;ter Meulen et al, 1981 ;Knobler et al, 1981Knobler et al, , 1982. Resistance to MHV-4-induced fatal central nervous system disease is inherited as a single autosomal recessive trait, independent of the major histocompatibility complex, expressed at the level of the neuronal cell (Knobler et al, 1981).…”