Persistence of mouse hepatitis virus A59 RNA in a slow virus demyelinating infection in mice as detected by in situ hybridization (original) (raw)
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Mouse hepatitis virus nasoencephalopathy is dependent upon virus strain and host genotype
Archives of Virology, 1986
Mouse hepatitis virus (MHV) S induced typical MHV spongiibrm lesions in brainstem 28 days following intranasal inoculation of adult A/J, BALB/ cByJ, CBA/J, C 3 H/HeJ and C 3 H/RV, but not SJL mice. In all but SJL mice, brain lesions occurred at or near the infectious dose level, based on seroconversion by the indirect immunofluorescence assay. During the acute phase of infection (day 5), lesions were limited to the nose and brain in most genotyoes. Exceptions were BALB mice, which had mild hepatitis and SJL mice, which had lesions restricted to the nose. No mortality occurred in any genotype. Following intranasal inoculation of adult mice, MHV-1, -3, -A 59, -JHM and -S all caused brain lesions at 28 days after inoculation. MHV-1 and -3 caused lesions that were usually restricted to the anterior offactory tracts, while MHV-A 59, -S and ~IHM also caused more generalized and pronounced lesions involving the midbrain and pons. These studies suggest that avirulent MHV-S given intranasally to most mouse genotypes is a good model for induction of brain infection in the absence of mortality. They also confirm observations made by others in which MHV-JHM, -S and -A 59 are relatively more neurotropic than other MHV strains, such as MHV-1 and -3.
In vitro replication of mouse hepatitis virus strain A59
Journal of Virology, 1987
An in vitro replication system for mouse hepatitis virus (MHV) strain A59 was developed using lysolecithin to produce cell extracts. In extracts of MHV-infected cells, radiolabeled UMP was incorporated at a linear rate for up to 1 h into RNA, which hybridized to MHV-specific cDNA probes and migrated in denaturing formaldehyde-agarose gels to the same position as MHV genomic RNA. The incorporation of [32P]UMP into genome-sized RNA in vitro correlated with the observed increase of [3H]uridine incorporation in MHV-infected cells labeled in vivo. Incorporation of [32P]UMP into genome-sized RNA was inhibited when extracts were incubated with puromycin. The addition to the assay of antiserum to the MHV-A59 nucleocapsid protein N inhibited synthesis of genome-sized RNA by 90% compared with the addition of preimmune serum. In contrast, antiserum to the E1 or E2 glycoproteins did not significantly inhibit RNA replication. In vitro-synthesized RNA banded in cesium chloride gradients as a ribo...
Journal of Virology, 2008
that mimics certain pathological features of multiple sclerosis. We have examined neural cell tropism of demyelinating and nondemyelinating strains of MHV in order to determine whether central nervous system (CNS) cell tropism plays a role in demyelination. Previous studies demonstrated that recombinant MHV strains, isogenic other than for the spike gene, differ in the extent of neurovirulence and the ability to induce demyelination. Here we demonstrate that these strains also differ in their abilities to infect a particular cell type(s) in the brain. Furthermore, there is a correlation between the differential localization of viral antigen in spinal cord gray matter and that in white matter during acute infection and the ability to induce demyelination later on. Viral antigen from demyelinating strains is detected initially in both gray and white matter, with subsequent localization to white matter of the spinal cord, whereas viral antigen localization of nondemyelinating strains is restricted mainly to gray matter. This observation suggests that the localization of viral antigen to white matter during the acute stage of infection is essential for the induction of chronic demyelination. Overall, these observations suggest that isogenic demyelinating and nondemyelinating strains of MHV, differing in the spike protein expressed, infect neurons and glial cells in different proportions and that differential tropism to a particular CNS cell type may play a significant role in mediating the onset and mechanisms of demyelination. on March 22, 2016 by guest http://jvi.asm.org/ Downloaded from 5520 DAS SARMA ET AL. J. VIROL. on March 22, 2016 by guest http://jvi.asm.org/ Downloaded from FIG. 4. Colocalization of EGFP-positive cells with astrocyte marker at day 5 postinfection.
2014
Neurotropic mouse hepatitis virus (MHV) is a member of the Coronavirus family induces hepatitis, meningitis, encephalitis, myelitis, neuritis, demyelination and axonal loss which mimics certain pathology of human neurological diseases multiple sclerosis (MS). Natural and genetically constructed recombinant MHV strains (generated by targeted RNA recombination) with differential pathological properties are used to understand the mechanisms of demyelination and concomitant axonal loss in a strain specific manner. These encompass both demyelinating (DM) and nondemyelinating (NDM) strains of MHVs, on which several comparative studies have been performed correlating the phenotypes, genomic sequences, and their pathogenicity. During acute infection, neurotropic DM strain infect a number of cell types in the CNS such as neurons, astrocytes, oligodendrocytes and microglia in the brain but in the spinal cord they preferentially infect neuron in the gray matter and oligodendrocytes and microgl...
Experimental and Molecular Pathology, 2001
Infection with mouse hepatitis virus (MHV) strain A59 produces and Cavanagh, 1997). Infections with nidoviruses, including acute hepatitis, encephalitis, and chronic demyelination in mice. Howboth coronaviruses and arteriviruses, have been used as ever, little is known about a closely related strain, MHV-2, which is experimental model systems for a variety of neurologic disonly weakly neurotropic. To better understand the molecular basis of neurotropism of MHVs, we compared the pathogenesis and genomic eases . MHV infection of mice is a useful sequence of MHV-2 with that of MHV-A59. Intracerebral injection of laboratory model system for virus-induced demyelination MHV-2 into 4-week-old C57B1/6 mice produces acute meningitis and that mimics many of the pathologic features of multiple hepatitis without encephalitis or chronic inflammatory demyelination. sclerosis Sequence comparison between MHV-2 and MHV-A59 reveals 94-98% sequence identity of the replicase gene, 83-95% sequence identity of Lavi et al., 1984a,b;; Houtman genes 2a, 3, 5b, 6, and 7, and marked difference in the sequence of and Fleming, 1996a,b). Studies of the neurotropic demyelingenes, 2b, 4, and 5a. This information provides the basis for further ating strains MHV-JHM and MHV-A59 provide important studies exploring the mechanism of viral neurotropism and virus-ininsight into the process of CNS infection; however, the mechduced demyelination. ᭧
Immunologic Research, 2007
Infection of mice with variants of mouse hepatitis virus, strain JHM (MHV-JHM), provide models of acute and chronic viral infection of the central nervous system (CNS). Through targeted recombination and reverse genetic manipulation, studies of infection with MHV-JHM variants have identified phenotypic differences and examined the effects of these differences on viral pathogenesis and anti-viral host immune responses. Studies employing recombinant viruses with a modified spike (S) glycoprotein of MHV-JHM have identified the S gene as a major determinant of neurovirulence. However, the association of S gene variation and neurovirulence with host ability to generate anti-viral CD8 T cell responses is not completely clear. Partially protective anti-viral immune responses may result in persistent infection and chronic demyelinating disease characterized by myelin removal from axons of the CNS and associated with dense macrophage/ microglial infiltration. Demyelinating disease during MHV-JHM infection is immunemediated, as mice that lack T lymphocytes fail to develop disease despite succumbing to encephalitis with high levels of infectious virus in the CNS. However, the presence of T lymphocytes or anti-viral antibody can induce disease in infected immunodeficient mice. The mechanisms by which these immune effectors induce demyelination share an ability to activate and recruit macrophages and microglia, thus increasing the putative role of these cells in myelin destruction.
Archives of Virology, 1993
To study the host-dependent genetic variations in murine hepatitis virus type 3 (MHV 3) induced diseases, we localized the sites of MHV 3 (Mill Hill strain) expression within liver and brain by immunohistochemistry or hybridization in situ. Two strains of mice were studied: BALB/c mice, which develop an acute and lethal hepatitis and C3H mice which develop a chronic brain infection. In BALB/c mice, viral RNA and antigens appeared during the first 24 h post infection (p.i.) in liver, whereas viral RNA was barely detectable in brain, up until death at day 3 p.i. In C3H mice, viral RNA and antigens were detected simultaneously in liver and brain only at day 2 p.i. In brain, the virus was detected in meningeal and ependymal cells and in perivascular cortical areas (days 5 and 7 p.i.). After day 49, the virus was no longer detected in brain parenchyma, but persisted in meningeal cells. Two host-dependent genetic differences in viral processing were observed in the liver: (1) the virus was first detected in Kupffer cells in BALB/c mice and mostly in hepatocytes in C3H mice; (2) in BALB/c mice, the 180kDa S viral glycoprotein appeared more frequently cleaved in 90 kDa form than in C3H mice.
Journal of Neuroimmunology, 1981
Demyelinadon may be induced by several different pathogenetic mechanisms. We have been utilizing mouse hepatitis virus (MHV) t o study virus-induced demyalination in the central net'vous system (CNS). To learn whether the different disease phenotypes in 4-week-old mice caused by wild type (a modal for fatal encephalomyditis) or mutaf.t 158 (a model for primary demyelination), is due to an altered cellular tropism, we have developed an immnnolabeling technique to evaluate critically the localization of MHV antigens in the unique ceils of the CNS. Using mouse derived L-cells and primar.~ neuronal cells in vitro, we determined an appropriate fixative (4% paraformaldehyde and 0.5% glutaraldehyde) that both preserved MHV antigenicity and cell structure. These studies in vitro showed the presence of MHV antigens on the surface of cells.