Mouse Macrophages as Host Cells for the Mouse Hepatitis Virus and the Genetic Basis of Their Susceptibility

Abstract

Much of the mystery of pathogenesis of animal virus disease lies in the realm of specificity of virus effect on cell type and in the differing effect of a given virus on different hosts. Tissue culture, despite its great contributions to other aspects of cytopathology, has not so far furnished significant clues to these basic problems. We have recently found that an acute virus disease of mice, mouse hepatitis," 2 has a selective destructive effect for cells derived from the reticulo-endothelial system (macrophages) and that the apparent genetic difference in susceptibility of different strains of mice is reflected in the behavior of macrophages from these strains in tissue culture. Tests of hybrids resulting from crosses between resistant and susceptible strains indicate that susceptibility is inherited and that genetic segregation of susceptibility and resistance occurs in the F2 and backeross generations.Materials and Methods.-The macrophages were obtained by explanting fragments of liver from newborn (1-3 days old) mice into roller tubes either directly onto the glass or onto a reconstituted collagen substrate.3' 4 The collagen was prepared according to the method of Ehrmann and Gey by extraction of 0.1 per cent acetic acid, dialysis against distilled water, and reconstitution to an agar-like slant with ammonium hydroxide vapors. The supernatant medium, except in otherwise specified cases; consisted of 60 per cent Gey's balanced salt solution, 10 per cent chick embryo extract (50 per cent), and 30 per cent horse serum (obtained from Microbiological Associates), with 0.004 per cent phenol red, 100 units of penicillin, and 10 micrograms of streptomycin. In some experiments, as designated in the text, chicken serum or a combination of chicken serum and horse serum was used. The chicken serum was obtained from White Leghorns kept for routine bleeding in our laboratory. The cultures were incubated in a roller drum and maintained in this manner at 370C throughout the experiments.The cultures were inoculated with the virus three to four days following explantation. The medium was renewed every two to three days until the end of the particular experiment. Supernatant fluids were frozen at -40'C and reserved for titration in mice. The cells were observed directly in the roller tube and their

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

“…C3H mice are resistant to MHV-2 (Bang & Warwick, 1960), A/J mice are resistant to MHV-3 (Virelizier & Allison, 1976) and SJL/J mice are resistant to MHV-4 (Stohlman & Frelinger, 1978;Knobler et al, 1981). Interferon has been implicated in the limitation of MHV-2 replication (Bang, 1981) and protects in vivo and in vitro against MHV-3 infection (Virelizier et al, 1977;Virelizier, 1981;Arnheiter & Hailer, 1981;Schindler et al, t982;Sorensen et al, 1982).…”

Studies on the Mechanism of Action of Riley Virus

“…The ability of the cells of the RES to phagocytose virus particles is well documented (15). Some viruses will replicate in macrophage cultures, for example mouse hepatitis virus (16,17), while others are phagocytosed without subsequently replicating, for example vaccinia virus (18,19). It is not possible to say at this stage what happens to Riley virus or the cells at the time of infection and during replication.…”

In Vitro Interaction of Mouse Hepatitis Virus and Macrophages From Genetically Resistant Mice

“…Methods of culture of mouse peritoneal macrophages have been described previously (2). Peritoneal washings were made 72 hr after the injection of sterile thioglycolate medium (Difco Laboratories, Inc., Detroit, Mich.) into the peritoneal cavity of mice.…”

“…The genetic resistance of C3H mice to infection with a mouse hepatitis virus (MHV) 1 grown in Princeton (PRI) mice (1) apparently resides to a large extent in the macrophage system, since macrophage cultures from the resistant mice are resistant in vitro while cultures from susceptible mice grown under identical conditions are susceptible (2,3). This can be tested by culturing macrophages from a group of mice and then backcrossing the mice whose macrophages are susceptible, to the inbred strain of C3H mice.…”