cord-000396-egy1d90x	2011	Prior studies using recombinant MHV strains that differ only in the spike gene, which encodes a glycoprotein involved in virus-host cell attachment, demonstrated that spike mediates anterograde axonal transport of virus to the spinal cord. A demyelinating MHV strain induces optic neuritis, but whether this is due to retrograde axonal transport of viral particles to the retina, or if it is due to traumatic disruption of retinal ganglion cell axons during intracranial inoculation is not known. The optic nerve inflammation, demyelination, and axonal loss observed after RSA59 infection, but not after RSMHV2 infection, demonstrate that the MHV-A59 spike protein is required for the induction of optic neuritis. This differential ability of MHV strains to induce optic neuritis and axonal injury is dependent on spike glycoprotein mediated retrograde transport of viral antigen along RGC axons.
cord-000409-lpf9lpky	2011	Fluorescence double-staining revealed that FGL2 and PD-1 were not co-expressed on the same cells, while quantitative RT-PCR demonstrated that higher levels of IFN-γ and TNF-α mRNA transcription occurred in PD-1-deficient mice in response to MHV-3 infection. In all, these results demonstrated that PD-1 deficiency led to enhanced pathological damage by MHV-3 in the liver, spleen, lymph node and thymus, where higher levels of PD-1-positive cells were found after infection. Results showed that the expression of FGL2 was principally associated with CD31-positive endothelial cells, CD68-positive macrophages and CD11c-positive DCs. Surprisingly, significantly higher levels of FGL2 were observed after infection in all of PD-1-deficient organs, including the liver, thymus, spleen, lymph nodes, and serum than that in those from WT littermates. However, the number of Foxp3-positive cells in the liver, spleen, lymph node or thymus was not significantly different between PD-1-deficient mice and their WT littermates after 72 h of MHV-3 infection (Fig. S3) .
cord-001017-4qfhltg4	2013	Previous studies demonstrated that microglia can be persistently infected by neurotropic mouse hepatitis virus (MHV) which causes meningoencephalitis, myelitis with subsequent axonal loss, and demyelination and serve as a virus-induced model of human neurological disease multiple sclerosis (MS). In our current studies, we have used RSA59 infection in vivo, in vitro, and ex vivo as a model to understand whether MHV can directly infect CNS resident microglia and the mechanism of microglial activation in the induction of chronic demyelination. To confirm the RSA59-induced CNS inflammation, brain and spinal cord sections from day 7 (peak of inflammation) and day 30 (peak of demyelination) postinfected mice were stained with H&E or LFB and examined. While Iba1 immunofluorescence was observed in both gray and white matter, double fluorescence/immunofluorescence demonstrated dual labelling of EGFP (viral antigen) positive Iba1 positive microglia/macrophages were present only in the white matter of RSA59 infected mice (Figure 1 ).
cord-001769-2sdg5ll7	2015	
cord-003199-03c9rx3o	2018	
cord-003378-0ozhye9q	2018	
cord-004663-a47pkh8q	1982	title: Ependymitis, leukoencephalitis, hydrocephalus, and thrombotic vasculitis following chronic infection by mouse hepatitis virus 3 (MHV 3) In semisusceptible mice, infection led first to a transient meningitis, ependymitis, and leukoencephalitis, followed by a permanent communicating hydrocephalus and, later on, to a chronic thrombotic vasculitis affecting meningeal and parenchymal vessels at the brain stem level. Identical lesions occurred in fully susceptible mice infected with a low dose of virus, but no neurologic disorder could be induced in genetically resistant mice even following immunosuppression or intracranial inoculation. When six susceptible BALB/c mice were injected i.p. with MHV 3 (103LD50), they died of an acute hepatic necrosis 5 -8 d a y s after M H V 3 infection, and no neuropatholigic lesion was observed except a slight degree of meningeal infiltration.
cord-004670-k1om7prn	1988	The mechanism of brain infection with mouse hepatitis virus-JHM was studied in BALB/cByJ mice following intranasal inoculation, and found to be a consequence of direct viral spread along olfactory nerves into olfactory bulbs of the brain. Lesions, antigen and virus were observed in the olfactory bulb and anterior brain as early as 2 days and posterior brain by 4 days after inoculation. The purpose of this study was to determine the route of virus entry into brain by examination of the sequential progression of infection with neurotropic MHV-JHM following i.n. inoculation of mice. To confirm that brain infection resulted from direct extension of virus along olfactory nerves and not due to viremia, groups of five mice were inoculated either i.n. or p.o. with MHV-JHM. Sequential studies of lesion, antigen and virus distribution in nose and brain indicated anterior to posterior progression of infection.
cord-004690-q38ogrem	1992	
cord-004728-rjl35dpa	1979	After intranasal inoculation of suckling rats mouse hepatitis virus multiplied mostly in the nasal epithelium; though there were no symptoms, antibodies were produced. Bar represents 0.1 mm Two-, 4-, 6-and t0-week-old rats were also inoculated i.n. with t05 PFU of MHV-S and infectious viruses were assayed in the lung, brain, salivary glands and liver. So far only mice have been considered to be natural hosts of MHV and rat; has been excluded because highly virulent MHV inoculated by various routes did not cause any sy~lptomatic infection like that produced in the mouse (7, 13) . B~tATT and his eoworkers showed that SDAV multiplied in mice producing some histopathological changes in the respiratory system after i.n. inoculation (3) and we demonstrated in the w e s e n t paper that 3{HV infected rats of any ages. Pathogenicity of mouse hepatitis virus for mice depending upon host age and route of infection
cord-004765-7e4yu2do	1992	
cord-007637-o2cijp5a	2005	In the present report we provide the strain distribution patterns of susceptibility to acute mouse hepatitis virus type-4 (MHV-4) encephalomyelitis, acute experimental allergic encephalomyelitis (EAE) and vasoactive amine sensitivity (VAAS) for 9 (CXJ) recombinant-inbred strains between BALB/cKe (C) and SJL/J (J) mice. Susceptibility to acute experimental autoimmune encephalomyelitis (EAE), an acute perivascular inflammatory demyelinating disease, is controlled by at least two genes, one, H-2 linked [responder haplotype (H-2 S, H-2 q or H-2d)]; the other associated with natural vasoactive amine sensitivity (VAAS) or that induced by Bordetella pertussis (Linthicum and Frelinger 1982) . Nine RI strains between BALB/cKe and SJL/J (CXJ) have been tested to provide the SDP of susceptibility to acute MHV-4 encephalomyelitis, acute EAE and VAAS. Finally, although some (CXJ) RI strains are susceptible to acute EAE, induced VAAS and acute M H V -4 encephalomyelitis EAE-like histopathological disease does not occur in these animals following MHV-4 intracerebral infection.
cord-009504-sn00p8iw	2013	The pathogenesis of mouse hepatitis virus (MHV‐S) infection in suckling and weanling mice was comparatively studied after intranasal inoculation. In the posterior part of the brain and spinal cord, virus was detected on days 3 to 4 postinoculation when viral growth was clearly demonstrable in the liver, spleen and intestines. In weanling mice, however, neither infectious virus nor viral antigen was detected in the liver or other visceral organs, while serum neutralizing antibody became detectable on day 5 postinoculation, increasing in titer thereafter. 2A and 2B , significant viral growth was observed in the brain, spinal cord ( Fig. 2A) and head without brain (Fig. 2B) , whereas no virus was demonstrated in the spleen or liver of the infected mice with a few exceptions (not included in the figures). In 4-week-old mice, however, no or little infectious virus was detected in the liver or other visceral organs, although high titered virus demonstrable in the brain was probably disseminated from the nasal mucosa as was observed in suckling mice.
cord-009793-t5bz4kmk	2005	
cord-009821-19dxy56e	2004	The present study examined the effects of MHV-JHM on cultured brain cells derived from Balb/c mice and from Wistar and from Lewis rats. To study the infection of astrocytes with MHV-JHM in vitro, cultures of brain cells derived from either newborn mice or rats were prepared and infected with virus. Infection of these oligodendrocyte-enriched cultures with MHV-JHM showed that 1 day after infection only a few GalC-positive (Fig. 4A,B) and no A2B5-positive cells (results not shown) contained viral proteins. When MHV-JHM infection caused an acute encephalitis in young rat pups, antigen could be found throughout the brain, mainly in astrocytes (Fig. 1A,B) . This susceptibility to MHV-JHM infection is found also in cultures of mouse astrocytes and Lewis rats (Massa et al., 1986) . Analysis of murine hepatitis virus (JHM strain) tropism toward Lewis rat glia cells in vitro: Type 1 astrocytes and brain macrophages (microglia) a s primary glial cell targets
cord-010252-go8cmgpo	2006	The hepatoprotection induced by synthetic muramyl peptides was investigated using a model of lethal murine mouse hepatitis MHV-3 virus infection. MDP and a nonpyrogenic analog, Murametide, inhibited the steep elevation of serum transaminases induced by MHV-3 irrespective of whether the immunomodulators were administered before or after the infection. In the present study, the effect of MDP and a nonpyrogenic analog, Murametide, on biochemical and other parameters was investigated using a model of lethal routine MHV-3 virus infection. In contrast, the MHV-3 infection induced increased GOT and GPT activities on day 3 and the enzyme levels were elevated even further on day 4, a time period when many of the animals were dying. Protective effect of muramyl dipeptide analogs in combination with trehalose dimycolate against aerogenic influenza and Mycobacterium tuberculosis infections in mice
cord-017613-va4ft5we	2005	The major receptor for murine coronavirus, mouse hepatitis virus (MHV), is identified as a protein, cell-adhesion molecule 1 in the carcinoembryonic antigen family (CEACAM1), which is classified in the immunoglobulin superfamily. This domain is also responsible for binding to the MHV spike (S) protein; the CC'' face protruding in this domain interacts with an N terminal region of the S protein composed of 330 amino acids (called S1N330). found that the plasma membranes isolated from MHV-susceptible BALB/c mouse hepatocytes or enterocytes contained a 110 to 120-kDa protein that binds to MHV particles, but those derived from MHV-resistant SJL mice lacked such a protein (2). Although CEACAM1 consisting of N domain alone bound MHV, it did not work as a functional receptor when expressed on CEACAMl-negative cells (18) . Communication between S1N330 and a region in $2 of murine coronavirus spike protein is important for virus entry into cells expressing CEACAM I b receptor
cord-019076-4qu9j953	2009	
cord-021413-1ht1xm88	2013	
cord-021499-up5vftj4	2007	
cord-022324-tcltmhi7	2012	
cord-022328-woktjl8h	2012	
cord-048204-6lvn10f4	2000	
cord-103306-1wc3f1rl	2020	Elevated mRNA and protein levels of tissue inhibitors of metalloproteinases 1 (TIMP-1) in MHV-A59 infection are suggestive of a TIMP-1 mediated host antiviral response. Total RNA was isolated from mock and virus-infected brain tissues 238 for expression analysis of viral nucleocapsid and Mmp genes through RT-qPCR. MMPs. To understand the regulation of MMPs upon MHV-A59 infection, we also 254 considered the gene expression of TIMPs. As described above, total RNA from brain samples 255 of mock and MHV-A59 infected mice were subjected to RT-qPCR using specific primers 256 12 (Table 1) to determine the transcript levels of Timp1, Timp2, Timp3, and Timp4. While Timp1 mRNA 260 followed a similar expression pattern as the Mmps following MHV-A59 infection-induced 261 inflammation, its protein levels remained high throughout post-infection, as shown in the 262 representative figure (Fig. 2, B) . Transcript levels of Parkinson''s disease 7 312 (Park7) gene were significantly upregulated following RSA59 infection and remained 313 elevated p.i compared to mock-infected samples (Fig. 7, A; p<0.05).
cord-104226-bb4lyvhy	1992	
cord-104253-v1r0idg0	1981	
cord-252882-2qhoqa88	2002	Targeted recombination was used to introduce amino acid substitutions into the cleavage signal of the fusion glycoprotein (spike or S protein) of MHV strain A59. The mutant cleavage and fusion phenotypes were seen only when the H716D substitution was present (Hingley et al, 1995; Leparc-Goffart et al, 1997 ; however, none of the glial cell mutants had the H716D mutation alone, so it was necessary to isolate recombinant viruses expressing the H716D mutation alone in order to assess the potential affect of this particular mutation on cleavage of S, fusogenicity, and virulence. Previous studies from this laboratory using mutants isolated from persistently infected glial cells (Gombold et al, 1993) have suggested that an H716D mutation, which changes the RRAHR cleavage signal to an RRADR sequence, is associated with an uncleaved S protein, but did not directly assess the affect of that mutation on virulence.
cord-253024-b393ea2u	1992	
cord-254871-qmx74umk	1987	Abstract Mouse hepatitis virus (MHV)-JHM infection was studied in genetically susceptible (BALB/cByJ) and resistant (SJL/J) mice following intranasal inoculation at 1, 3, 6 or 12 wk of age. Mouse hepatitis virus (MH~-JH~ infection was studied in genetically susceptible (BALB/cBy~ and resistant (SJL/J) mice follo~ng intranasal inoculation at 1, 3, 6 or 12 wk of age. The following tissues were specifically examined for MHV antigen and lesions in mice of both genotypes, all ages and all intervals: nose, eye, brain, spinal cord, lung, liver, spleen, submaxillary and mesenteric lymph nodes, salivary glands, bone and bone marrow, small intestine, cecum, colon, kidney, urinary bladder and gonad. Lesions and antigen of 1-wk-old SJL mice resembled BALB mice, and were found in nose, olfactory bulb, brain, liver, spleen, lymph nodes, bone marrow and gut associated lymphoid tissue (Fig. 1 ). Titers of MHV-JHM were determined in brain, liver, spleen and intestine at 0, 3, 5,10,20 and 30 days after inoculation of Gwk-old BALB and SJL mice (Fig. 8) .
cord-256149-btjq84q7	2006	Synthetic decapeptides (N = 206) covering the entire sequence of mouse liver fumarylacetoacetate hydrolase (FAH) were used to analyze the specificities of the autoantibodies (autoAb) elicited towards this enzyme in mice infected with mouse hepatitis virus (MHV). Results indicated that the induction of the autoAb is not only related to molecular or structural mimicry, but rather supports the Danger model, in which any aggression, in this case the MHV infection, is susceptible to trigger the production of autoAb. Mouse hepatitis virus strain A59 (MHV-A59) is a coronavirus that triggers various pathologies in susceptible mice, including hepatitis and thymus involution, IgG2a-restricted hypergammaglobulinaemia and transient demyelination [1, 2] . Overlapping decapeptides corresponding to the entire mouse FAH sequence were prepared using the PEPSCAN method and their reactivities with sera from MHV-infected mice at different times was determined by ELISA. Results indicated that various regions of the enzyme, including sequence 1e20, are recognized as soon as 15 days after infection and that the autoimmune response is not restricted to peptides homologous to viral proteins.
cord-256444-grw5s2pf	1997	Publisher Summary This chapter discusses the manipulation of clones of coronavirus and of complementary DNAs (cDNAs) of defective-interfering (DI) RNAs to study coronavirus RNA replication, transcription, recombination, processing and transport of proteins, virion assembly, identification of cell receptors for coronaviruses, and processing of the polymerase. This decade has seen the manipulation of these clones, and of complementary DNAs (cDNAs) of defective-interfering (DI) RNAs, to study coronavirus RNA replication, transcription, recombination, processing and transport of proteins, virion assembly, identification of cell receptors for coronaviruses, and processing of the polymerase. The species and tissue specificity of a coronavirus infection is a t least partially dictated by the nature and distribution of cellular receptors and other related molecules that regulate virus entry, as evidenced by the viral replication that results when viral RNA is directly introduced into cell types of other animal species.
cord-258286-lodjcj8c	1997	Previous studies have shown that immunocoma large plaque variant derived from the parental JHM petent mice infected with MHV exhibited increased exstrain (Stohlman et al., 1982) ; the small plaque variant pression of a number of cytokines, including IL-1, IL-6, DS (Stohlman et al., 1982) ; the neutralization-escape mu-TNF-a, and IFN-g, in the CNS at the time of viral cleartant 2.2-V-1 (Fleming et al., 1987; Wang et al., 1992) , and ance (Pearce et al., 1994) . The expressed IFNg exhibited antiviral activity, prevented virus spread in layers of DBT cells grown at approximately 70% confluence in 60-mm petri dishes were infected with MHV at vitro, and altered viral pathogenesis in mice. Expression of IFN-g using an MHV DI RNA vector cell metabolism prior to infection or it may be that interferon acts at an early stage of viral replication.
cord-259095-mfptcw8t	1997	
cord-259374-m7q1roay	2019	
cord-259580-fn5pkec9	2020	
cord-259603-bh198xgl	2016	
cord-259671-7de21oaq	2014	Overall, the conservation pattern identified for 5′ and 3′-terminal RNA structural elements in the genomes of alphaand betacoronaviruses is in agreement with the widely used replicase polyprotein-based classification of the Coronavirinae, suggesting co-evolution of the coronavirus replication machinery with cognate cis-acting RNA elements. Other internal cis-acting elements include specific RNA signals required for genome packing, which have been characterized in a small number of coronaviruses Escors et al., 2003; Morales et al., 2013; Penzes et al., 1994) , and a complex RNA pseudoknot structure located in the ORF1a-ORF1b overlap region that mediates a (−1) ribosomal frameshift event and thus controls the expression of the second large ORF on the coronavirus genome RNA (ORF1b) (Brierley et al., 1987 (Brierley et al., , 1989 de Haan et al., 2002; Namy et al., 2006) .
cord-260177-xu0elmak	1982	
cord-261291-0lntii22	1985	Abstract A panel of 28 monoclonal antibodies (MAb) against the structural proteins of murine hepatitis virus-4, strain JHM (MHV-4) was used in three antigen binding assays to determine the extent of antigenic homology among six strains of murine coronaviruses. Antigenic preparations for dot immunoblotting and enzyme immunoassays (EIA) consisted of microsomal fractions from MHV infected or uninfected control cells, as described previously (Talbot et al., 1984a) . (1983) , extensive conservation of the El glycoprotein was evident and similar levels of antibody binding were seen on each MHV strain, with the exception of the ts8 mutant of MHV-4. 28 monoclonal antibodies against the JHM strain of murine hepatitis virus (MHV-4) were used in three different antigen binding assays to delineate antigenic relationships among the structural proteins of six MHV strains.
cord-262505-1ufgwxxg	1983	
cord-263658-dlmzcl9g	1970	authors: MCINTOSH, KENNETH; KAPIKIAN, ALBERT Z.; TURNER, HORACE C.; HARTLEY, JANET W.; PARROTT, ROBERT H.; CHANOCK, ROBERT M. Table 1 shows the proportion of infants and children with and without LRTD showing fourfold or greater antibody responses to the three related coronavirus antigens OC38, OC43, and MHV, strain A-59. It is evident from the studies of hospitalized children that infection with coronaviruses was not significantly associated in this survey with pediatric lower respiratory tract disease. It is of interest that in the seroepidemiologic studies of Hartley and others, where several strains of MHV were tested with sera from military recruits, no epidemiologic association of MHV or MHV-like virus infection with respiratory tract disease was made (11) . Antigenic studies of human coronaviruses have shown that those members of the group which were originally recovered in tissue culture are all closely related to the prototype virus strain 229E (2, 5) .
cord-263678-z94utwbk	2004	The authors have previously shown that the spike (S) glycoprotein gene of MHV contains determinants of virulence, hepatitis, and demyelination. In the present study, the authors produced new recombinant viruses with each one of these S gene mutations by site-directed mutagenesis and targeted recombination and studied the effect of each individual mutation on the pathogenesis of the virus. The use of the virus with feline spike protein (fMHV) also provided a Coronavirus S gene demyelination determinants L Fu et al 43 useful method to select recombinant viruses against the background of parental viruses. Viral RNA persistence of all recombinant and parental viruses in mouse organs (brain, spinal cord and liver) was analyzed by reverse transcriptase L Fu et al Figure 3 Pathologic analysis of recombinant viruses during acute infection. Targeted recombination within the spike gene of murine coronavirus mouse hepatitis virus-A59: Q159 is a determinant of hepatotropism
cord-264848-wl29jk16	2004	Transplantation of glial-committed progenitor cells into the T8 spinal cord of MHV-infected mice demonstrating complete hindlimb paralysis resulted in migration of cells up to 12 mm from the implantation site and remyelination of up to 67% of axons. Counts of the total number of axons (normally myelinated, demyelinated, and remyelinated) within the region extending 8 mm cranial and 6 mm caudal to the transplantation site (the extent of spinal cord examined) suggest that transplanted animals had significantly more axons ( P < 0.01) within the ventral and lateral columns as compared to non-transplanted animals (Figs. Multipotential PSA-NCAM + neural precursors isolated from the postnatal rat brain have been shown to differentiate into oligodendrocytes, Schwann cells, and astrocytes following transplantation, to completely remyelinate regions of acute demyelination in the adult rat induced by ethidium bromide injection into x-irradiated dorsal column white matter .
cord-264884-ydkigome	2008	
cord-265895-ck7eto16	1987	These data, coupled with the high frequency of RNA recombination during MHV infection, suggest that the viral polymerase may pause in or around regions of secondary structure, thereby generating pools of free leader-containing RNA intermediates which can reassociate with the template, acting as primers for the synthesis of full-length or recombinant RNAs. These data suggest that MHV transcription uses a discontinuous and nonprocessive mechanism in which RNA polymerase allows the partial RNA products to be dissociated from the template temporarily during the process of transcription. These data, coupled with the presence of discrete large leader-containing RNAs which range from 84 to 1000 nucleotides in length in MHV-infected cells (Baric et a/., 1985) suggest that discontinuous RNA intermediates may be dissociated and reassert between viral RNA templates to generate recombinant viruses by a copy-choice mechanism (Makino eta/., 1986a). The leader-containing RNAs larger than 1 10 nucleotides in length detected by the leader-specific cDNA probe were more heterogeneous (Fig. 2) (Baric et al., 1985) suggesting that multiple RNA species in this size range were present.
cord-266018-8bhnlsgy	2004	
cord-266138-yibbiiij	1995	Murine coronaviruses (mouse hepatitis virus, MHV) can spread inapparently or may hide as persistent infections that modulate the immune response [38, 100] . Suppression of immune response induction in Peyer''s patch lymphoid cells from mice infected with mouse hepatitis virus Infection of BALB/cByJ mice with the JHM strain of mouse hepatitis virus alters in vitro splenic T cell proliferation and cytokine production Interaction of immune and central nervous systems: contribution of anti-viral Thy-1 + cells to demyelination induced by coronavirus JHM Impaired T and B cell subpopulations involved in a chronic disease induced by mouse hepatitis virus type 3 Identification of antigenic sites mediating antibody-dependent enhancement of feline infectious peritonitis virus infectivity The pathogenic role of virus-specific antibody-secreting cells in the central nervous system of rats with different susceptibility to coronavirus-induced demyelinating encephalitis Demyelination induced by murine hepatitis virus JHM strain (MHV-4) is immunologically mediated
cord-267671-ys43n672	2015	Clinical Signs MCMV causes subclinical infection in adult immunocompetent mice, but experimental inoculation of neonates can cause lethal disease due to multisystemic necrosis and inflammation. Diagnosis Because infected mice do not manifest signs or lesions and the virus is very difficult to propagate in cell culture, detection and diagnosis rely on serology and molecular methods. Differential Diagnosis Reovirus infection must be differentiated from other diarrheal diseases of infant mice, including those caused by mouse coronaviruses, EDIM virus, Salmonella spp., or Clostridium piliforme. Epizootiology EDIM virus appears to be infectious only for mice and occurs episodically in mouse colonies, and infection is probably widespread geographically (Livingston and Riley, 2003; Pritchett-Corning LABORATORY ANIMAL MEDICINE et al., 2009) . Sentinel mouse surveillance, using soiled bedding, is an effective strategy for detecting MNV (Manuel et al., 2008) Differential Diagnosis The mild change in fecal consistency associated with MNV in adult mice may mimic rotavirus, coronavirus, Helicobacter spp., Citrobacter rodentium, or other enteric diseases.
cord-268139-tgpsu4qz	2005	title: Mutagenesis of the murine hepatitis virus nsp1-coding region identifies residues important for protein processing, viral RNA synthesis, and viral replication In the current study, a reverse genetic approach was used to define the role of the 28-kDa amino-terminal product (nsp1) of the gene 1 polyprotein during replication of the coronavirus murine hepatitis virus (MHV) in cell culture. To determine whether nsp1 is required for MHV replication and to identify residues critical for protein function, mutant viruses that contained deletions or point mutations within the nsp1-coding region were generated and assayed for defects in viral replication, viral protein expression, protein localization, and RNA synthesis. Mutagenesis of the murine hepatitis virus nsp1-coding region identifies residues important for protein processing, viral RNA synthesis, and viral replication Experiments were next performed to determine if viral gene expression occurred in cells electroporated with RNA for mutants that did not produce infectious virus (VUSB5, VUSB6, nsp1DFL, and nsp1DMid).
cord-268238-ipfs7hcb	2004	
cord-268416-8hw80qx8	2018	While screening for ADP-ribosylated proteins during coronavirus (CoV) infection, we detected a ~55 kDa ADP-ribosylated protein in mouse hepatitis virus (MHV)-infected cells and in virions, which we identified as the viral nucleocapsid (N) protein. To identify potential targets of the CoV macrodomain, we analyzed infected cells for changes in ADP-ribosylation patterns utilizing antibodies specific for ADPr. We focused on cells infected with a murine CoV, mouse hepatitis virus (MHV). If the macrodomain was indeed removing the ADPr from the N protein, we would expect to see increased N protein ADP-ribosylation in cells infected with mutant virus compared to wild-type infected cells. To determine if N protein expressed in the absence of CoV infection could be ADP-ribosylated in cell culture, we transduced Vero cells with VEEV replicon particles (VRPs) encoding the MERS-CoV N protein or control GFP at different MOIs .
cord-269011-230p8rsf	2005	Biochemical and theoretical studies led to a topological model for the MHV M protein Rottier et al., 1984 Rottier et al., , 1986 , in which the polypeptide spans the lipid bilayer three times, leaving a small amino-terminal domain (15-35 residues) in the lumen of intracellular organelles (or outside the virus), whereas the carboxyterminal half of the protein is located on the cytoplasmic side of the membrane (or inside the virion). Using a mutational approach including deletions, insertions, and substitutions, both the transmembrane domain and the cysteine-rich region immediately downstream thereof, but not the carboxy-terminal part of the cytoplasmic tail, were shown to be important for MHV S protein-induced cell-cell fusion (Bos et al., 1995; Chang and Gombold, 2001; Chang et al., 2000) The cleavage requirements of the S proteins for the biological activities of the coronavirus spike remain enigmatic.
cord-270473-5tok4mqk	2003	We have previously shown that mitochondrial-aconitase binds specifically to the 3′ terminal 42 nucleotides of the Murine hepatitis virus (MHV) RNA along with three additional proteins of 70, 58 and 40 kDa to form a stable RNA-protein complex. The assays were done essentially as previously described [22] except that the partially purified post-mitochondrial lysates (approximately 3 µg total protein) were incubated at 22 • C with purified m-apo-aconitase (8-10 µg, graciously supplied by M.C. Kennedy, Gannon University, Erie, PA) and MHV-JHM 3 UTR RNA in ATPase buffer (20 mM Hepes, pH 7.4, 100 mM KCl, 5 mM MgCl 2 , 0.1% NP-40, 1 mM DTT, 1 mM ATP containing 2 µCi [γ-32 P]ATP plus cocktails of protease and phosphatase inhibitors). Gel supershift assays with anti-phosphotyrosine antibodies (Fig. 2B ) demonstrate that at least one of the proteins in the complex formed with MHV 3 (+)42 containing RNAs and m-aconitase, mtHSP70, HSP60, and HSP40 is tyrosine phosphorylated.
cord-270814-krw8zmr5	1997	
cord-271526-14nfqusv	1997	
cord-271763-cual2qv4	1990	Abstract The sequence of the spike (also called peplomer or E2) protein gene of the Mebus strain of bovine coronavirus (BCV) was obtained from cDNA clones of genomic RNA. Amino terminal amino acid sequencing of the virion-derived gp100 spike subunit confirmed the location of the predicted cleavage site, and established that gp120 and gp100 are the glycosylated virion forms of the S1 and S2 subunits, respectively. The deduced amino acid sequence of the extended ORF demonstrated high sequence similarity to the C-terminal end of the antigenically related MHV-A59 (22) and MHV-JHM (32) S proteins, both antigenic homologs of the BCV S protein (13). The N-terminal sequence of the lOO-kDa subunit could be obtained, however, and was determined to be X-l-T-T-G-Y-X-F-, identifying the first amino acids downstream from the predicted internal cleavage site.
cord-271815-yr1dq258	2011	
cord-274247-5qiwui6u	1999	We have investigated the effects of a natural acute outbreak of MHV in our otherwise specific-pathogen-free (SPF) inbred mouse colonies, and of enzootic chronic MHV infection on cytokine production and resistance to the intracellular pathogen Trypanosoma cruzi. IL-10 that were serologically MHV+ or MHV− production, on day 11 after infection, in BALB/c (but not in C57BL/6) mice was CD4-activation dependent as treatment Our preliminary observation, suggestive of an infection with GK1.5 mAb suppressed most of its synthesis outbreak in the mouse colonies, was increased susceptibility (63 units/ml in untreated versus 6 units/ml in GK1.5 treated to T. cruzi determined by the situation of acute MHV+ C57BL/6 and BALB/c mice cells to Con A stimulation simultaneous infection with blood parasites derived from was not significantly higher than in spleen cell cultures from acutely infected MHV+ donors as described above, no statis-MHV− animals (Fig. 2) .
cord-276198-psjua913	2015	Many nidovirus and all coronavirus TM1 proteins contain one or more ubiquitin-like domains which may help to anchor the viral RNA to the membranes where replication takes place (Hurst et al., 2013) . The maze-like paired-membrane structures that resulted from coexpression of SARS-CoV TM1 and TM2 have not ever been reported in coronavirus-infected cells, suggesting that this should be interpreted as a conditional, or perhaps partial phenotype, that is not observed when the full viral replicase polyprotein is expressed. Since DMVs are reminiscent of the double-membranes of autophagosomes, several lines of controversial evidence hypothesized a diversion of Atg (autophagyrelated) proteins and autophagosome function during coronavirus replication, as it is the case for other +RNA viruses (Prentice et al., 2004; Snijder et al., 2006; Zhao et al., 2007; Maier and Britton, 2012; Richards and Jackson, 2013) . Expression of hepatitis C virus proteins induces distinct membrane alterations including a candidate viral replication complex
cord-277566-j3ehiwn9	2008	
cord-281552-zfjy3m3i	2020	Here, we test E-derived peptides for membrane binding activity in vitro and confirm those identified as positive in the context of the full length protein expressed in two different cell types. Relative densitometry of the HSP and LSP bands revealed significant differences among the mutants with regard to their localisation to the different membrane fractions of E expressing insect cells ( Figure 5B ) and confirmed a role for the amphipathic MHV CoV E 50-64 peptide in membrane interaction. An amphipathic helix, EPTM, detected in the post-TM region of E, was suggested by bioinformatics analysis and assessed for direct membrane interaction in vitro by binding to GUVs. For comparison, the predicted E protein TM domain, ETM, and an established membrane active peptide from the influenza M2 protein were also included. Following expression of the complete E protein with mutations in the same identified peptide, altered membrane binding in two distinct cell types, mammalian and insect, was apparent.
cord-283132-rfw8njpo	1993	
cord-284707-72vx11aq	1988	For mouse hepatitis virus (MHV), one of the most extensively studied coronaviruses, there are seven species of MHV-specific mRNA present in infected cells, the largest of which is indistinguishable from virion RNA (Leibowitz et a/., 1981; Lai et a/., 1981; Spaan et a/., 1981) . In this paper we report the characteristics of a permeabilized cell system and demonstrate that it incorporates ribonucle-otide triphosphates into RNA molecules which appear identical to the virus-specific mRNAs synthesized in MHV-infected cells. Protease inhibitors and RNase inhibitors have both been reported to increase the RNA-dependent RNA polymerase activity present in extracts of West Nile virus-infected cells (Grun and Brinton, 1986) . The kinetics of the development of the MHV-specific RNA polymerase activity over the course of infection was determined in permeabilized cells. In this work we report the development and characterization of a permeabilized cell system for assaying MHV-specific RNA polymerase activity.
cord-285676-4kgy20o9	1997	The overlapping ORFs 1a and b found at the 58 end of the nidoviral genome are frequently referred to as the ''''polymerase gene.'''' However, there is little doubt that the processing of the encoded polyproteins yields proteins required for RNA synthesis as well as a number of products involved in other aspects of virus replication. The sequence conservation between the more closely related corona-and toroviruses is clustered in six domains, four of which are also found in the arterivirus POL1b: the ''''classical'''' RNA-dependent RNA polymerase (RdRp) and helicase (H) domains, which are also present in the polymerases of most other viruses, a zinc finger motif (zf), and a short region of 80-100 residues, which has not yet been identified in other viral polymerases and was called the ''''coronavirus-like'''' (CVL) domain (3) (motif 2 in Fig. 1b) .
cord-285869-jwflooop	2008	
cord-286250-bq1u3d4z	2001	
cord-286332-cdg4im5h	2017	
cord-287487-qeltdch7	2017	
cord-289045-vft163v0	2005	
cord-290877-dap0zo2m	2018	
cord-292178-bd8u8udl	1993	title: Interleukin-6 induction in vitro in mouse brain endothelial cells and astrocytes by exposure to mouse hepatitis virus (MHV-4, JHM) Abstract Interleukin-6 (IL-6) induction, as detected by bioassay and Northern analysis, was examined in vitro in endothelial cells or astrocytes derived from BALB/c (susceptible) or SJL (resistant) mice following exposure to mouse hepatitis virus (MHV-4) or UV inactivated MHV-4 (UV-MHV-4). BALB/c (MHV-4 susceptible) derived endothelial cells can produce up to 16-fold higher levels of IL-6, and release this earlier than SJL (MHV-4 resistant), as determined both by bioassay and Northern analysis. Tables 1 and 2 demonstrate that IL-6 is induced in both BALB/c and SJL derived endothelial ceils following exposure to MHV-4 as determined by bioassays, albeit at very different levels. We report on the induction of IL-6 in cultures of cerebral endothelial cells or astrocytes following infection with MHV-4.
cord-292424-daj4zcm1	2020	Together these results reveal a new role of receptor binding in MHV entry: in addition to its well-characterized role in viral attachment to host cells, receptor binding also induces the conformational change of the spike and hence the fusion of viral and host membranes. In this study, we determined the cryo-electron microscopic (cryo-EM) structure of pre-fusion MHV spike in complex with CEACAM1a (D1-D4), which reveals the structural change of MHV spike associated with receptor binding. Using proteolysis and negative-stain EM assays, we further investigated the impact of receptor binding on proteases sensitivity and the final structural transitions of MHV spike. The result showed that receptor treatment of the trypsin-cleaved MHV S-e led to a protease K-resistant S2'' fragment, suggesting that CEACAM1a binding facilitated the already cleaved MHV S-e to transition from pre-fusion to post-fusion conformation.
cord-293417-oqusfhei	2010	The higher resolution provides more detailed structural information than previous reports, showing that the NTD structure from MHV shares a similar overall and topology structure with that of SARS-CoV and IBV, but varies in its potential surface, which indicates a possible difference in RNA-binding module. To gain insight into the precise mechanism of N protein, several crystallographic or NMR structural results were reported, including MHV N-terminal RNA binding domain (residues 60-195) (Grossoehme et al., 2009) , two proteaseresistant domains of the N protein from SARS-CoV (Huang et al., 2004; Luo et al., 2006; Yu et al., 2006; Chen et al., 2007; Saikatendu et al., 2007; Takeda et al., 2008) , and IBV (Beaudette strain and Gray strain) (Fan et al., 2005; Jayaram et al., 2006) . In overall ribbon posture, the high resolution structure of MHV-NTD determined using two forms of crystals with different packing modes is similar to previously reported SARS-CoV and IBV structures, with a remarkable difference in surface electrostatic distribution.
cord-293913-frkb8iso	1996	
cord-293975-np9xdag5	1993	The data also demonstrate that two viruses can enter the brain via the olfactory system and localize to different structures, suggesting that neurological diseases involving disparate regions of the brain could be caused by different viruses, even if entry occurred at a common A number of viruses have been shown to spread transneuronally into and throughout the rodent CNS following intranasal inoculation, including herpes simplex virus type 1 (HSV-I)," vesicular stomatitis virus,43 Borna disease virus," mouse hepatitis virus (MHV),-pseudorabies viru? Abbreviafions: HSV-1, hernes simplex virus, type 1; LC, locus coeruleus; MHV''-JHM, -mouse hepatitis virus, strain JHM; MOB, main olfactorv bulb; PFU, plaque forming unit; p.i. post-inoculation; TH, tyrosine hydroxylase; TH + , tyrosine hydroxylase immunoreactive; TH -, tyrosine hydroxylase immunonegative; VTA, ventral tegmental area; WGA-HRP, wheatgerm agglutinin-horseradish peroxidase.
cord-294467-kq5wmavt	2014	title: Characterization of haemagglutinin-esterase protein (HE) of murine corona virus DVIM by monoclonal antibodies We analyzed the characteristics of seven monoclonal antibodies (mAbs) raised against purified HE (hemagglutinin-esterase) glycoprotein of the murine coronavirus DVIM (diarrhea virus of infant mice). Neutralizing activity was shown in group A mAbs exclusively, suggesting that the ratio of HA and/or AE activities may play important roles in the cell fusion activity of DVIM-infected cells. As described in a previous report [8] , by treatment of DVIM with the non-ionic detergent NP-40 we isolated the HE protein, which carries the functional sites for both acetylesterase (AE) and the haemagglutination (HA) activities. We examined the cytopathic effect of mAbs with respect to the formation of syncytia in DVIM-infected cells. The antibodies that had low titers to both activities did not delay the fusion of infected cells.
cord-295307-zrtixzgu	2020	Through the integration of differential expression analyses and reconstructed networks exploration, significant differences in the immune response to virus were observed in Ly6E [Formula: see text] compared to wild type animals. Among the different types of GNs, gene co-expression networks (GCNs) are widely used in the literature due to their computational simplicity and good performance in order to study biological processes or diseases [8] [9] [10] . In the present work mice samples were compared organ-wise depending on whether these corresponded to control, 3 d p.i. and 5 d p.i. The identification of DEG was performed using the Limma [63] R package, which provides non-parametric robust estimation of the gene expression variance. In this work four gene networks were reconstructed to model the genetic response MHV infection in two tissues, liver and spleen, and in two different genetic backgrounds, wild type and Ly6E ∆HSC .
cord-295781-b831y105	2017	title: Lung epithelial cells have virus-specific and shared gene expression responses to infection by diverse respiratory viruses We used microarray analysis to compare changes in gene expression of murine lung epithelial cells infected individually by three respiratory viruses causing mild (rhinovirus, RV1B), moderate (coronavirus, MHV-1), and severe (influenza A virus, PR8) disease in mice. While these models are invaluable for evaluating pathology and host responses to infection, parallel in vitro studies can be used to identify gene expression and signaling pathway changes that occur in infected cells to mediate pathogenesis. In this study, we compare the gene expression response of murine lung epithelial cells to infection by three respiratory viruses used in murine models: rhinovirus (serotype RV1B) in the family Picornaviridae, mouse hepatitis virus (MHV strain 1) in the family Coronaviridae, and influenza A virus (strain PR8) in the family Orthomyxoviridae.
cord-296416-q0rsfzgw	1996	
cord-298326-f5q7j3iu	2020	In this study, we describe the identification of an essential, conserved horseshoe-shaped region in the nsp5 interdomain loop (IDL) of mouse hepatitis virus (MHV), a common coronavirus replication model. Structural modeling and sequence analysis of these sites in other coronaviruses, including all 7 human coronaviruses, suggests that the identified structure and sequence of this horseshoe regions is highly conserved and may represent a new, non-active-site regulatory region of the nsp5 (3CLpro) protease to target with coronavirus inhibitors. To evaluate whether any of 207 the recovered MHV nsp5 IDL mutants may exhibit a temperature-sensitive phenotype, we 208 performed an efficiency of plating (EOP) analysis by comparing the titers of each IDL virus by 209 plaque assay determined at a physiologic (37°C) and elevated temperature (40°C) (Fig. 3A) .
cord-298847-szezd2vb	2003	
cord-298934-vtrfqozl	1988	
cord-299756-m0va36er	2009	
cord-301942-ppa7gb95	2006	The current understanding of coronavirus ultrastructure relies heavily on transmission electron microscopy of negatively stained images. Each virus appeared approximately round in cryo-EM images, with a fringe of spikes protruding from the viral membrane and a region of lower density near the virion center ( Fig. 1A-B) . Spike-depleted SARS-CoV particles appeared similar to spike-depleted MHV particles in negative stain, but were produced in lower yield, not suitable for effective cryo-EM imaging. The arrangement of spike densities near the center of some particles approximates a rhombus, which would not be inconsistent with a paracrystalline organization of spikes as observed in the virions of pleomorphic arenavirus particles, 17 or a local hexagonal close-packing of structural proteins as observed in retroviral particles. Fine structure of influenza A virus observed by electron cryo-microscopy Cryo-electron microscopy reveals ordered domains in the immature HIV-1 particle Supramolecular organization of immature and mature murine leukemia virus revealed by electron cryo-microscopy: implications for retroviral assembly mechanisms
cord-303153-z7bdiuvx	2010	This approach has allowed us to establish that MHV induces the formation of six membranous rearrangements in the following order: DMVs, CMs, virions, LVCVs, TBs and CMSs. Importantly, we were able to show that most membrane rearrangements (LVCVs, TBs, CMSs and possibly CMs) observed in addition to the key structures in the infection (DMVs and virions) actually appear to be the consequence of a massive synthesis of viral proteins. In order to be able to correlate our EM and IEM analyses with the progression of a CoV infection inside the host cells, we first measured important known parameters that reflect the CoV life cycle: viral RNA replication/transcription, viral protein synthesis and secretion of progeny virus. In the centre of the DMV clusters, we frequently observed a small network of membranes with a diameter varying from 200 to 600 nm, which have recently been described in SARS-CoV-infected cells and called CMs [ Fig. 2A and B, arrowheads; (Knoops et al., 2008) .
cord-303238-us3dybue	2007	The papain-like protease (PLpro) encoded by the coronavirus that causes severe acute respiratory syndrome (SARS-CoV) processes three sites in the replicase polyprotein (Harcourt et al., 2004) , and has recently been shown to have de-ubiquitinating activity (Barretto et al., 2005; Lindner et al., 2005) . To extend these studies of membrane association of coronavirus replicase products, we analyzed the amino acid sequence of MHV-JHM nsp3 (from glycine-833 to glycine-2840) for probability of transmembrane helices using the five different programs designed to search for putative membrane-spanning sequences: Phobius, TMHMM, HMMTOP, SOSUI and TMpred (Fig. 2) . In contrast, when CMMs were added to the mixture, protein products that included all or part of nsp3-TM (PLP2-2485, -2390 and -2258) were detected predominantly in the pelleted fraction, consistent with membrane association (Fig. 3B ).
cord-304421-xpj6c0vx	1999	
cord-304855-7v0cncid	2009	
cord-304954-5b4yji8n	1991	title: Production of mice from a lethal coronavirus infection in the central nervous system by adoptive transfer of virus-specific T cell clones Abstract The protective effect of a mouse hepatitis virus type-4 (MHV-4)-specific CD8+ cytotoxic T cell clone and a CD4+ helper T cell clone was examined by the adoptive transfer into brains of mice lethally infected with MHV-4. Although the adoptive transfer of both types of the T cell clones suppressed viral growth and viral antigen-positive cells in the brains, a significant inhibition of virus replication by the cytotoxic T cell clone was detected prior to that induced by the helper T cell clone. In the present study we demonstrate that the adoptive transfer of virus-specific CD8 ÷ CTL clones also protects mice from a lethal MHV-4 infection. In this study we have confirmed that the adoptive transfer of virus-specific CD4 + Th cells protects mice from a MHV-4 lethal infection.
cord-307098-oq7zrnuv	2014	Co-cultivation of MHV-nonpermissive hamster BHK cells devoid of mouse Bgp with mouse DBT cells infected with MHV-A59 or JHMV induces syncytia formation on BHK cells (Bgp-independent fusion). Under a phase contrast microscopy, JHMV (cl-2, sp-4) induced the Bgp-independent syncytia on BHK cells similar to those observed on DBT cells, while such syncytia were not seen with the infection of other MHV strains (MHV-1, MHV-3, MHV-A59, MHV-S, srr7, srr11 and srr18). Interestingly, no syncytia formation was demonstrated on BHK cells with other strains or srr mutants under the phase contrast microscopy, though all of them produced syncytia on Bgp-positive DBT cells [10] . The cl-2 S protein could have a very strong Bgp-independent fusion activity, but its replication in BHK cells could be less efficient than the other MHV strains. Present study showed that JHMV (cl-2 and sp-4) could induce syncytia on BHK cells detectable by microscopy, whereas the other MHVs and srr mutants failed.
cord-309109-c5hajb6k	2002	Although activated , MHV-speci c, CD8 C T cells transferred to infected mice can control viral replication, they are not suf cient to clear infectious virus from oligodendrocytes (Stohlman et al, 1998b) . Clearance of an MHV strain that primarily infects neurons is delayed in the absence of IFN-gamma, suggesting that it may affect viral control in other cell types as well without being absolutely required for clearance (Lane et al, 1997) . The contribution of humoral immunity to viral clearance and persistent infection in the CNS has been investigated using mice de cient in secreted antibodies or B cells Bergmann et al, 2001; Matthews et al, 2001) . These data suggest that T cells play a role in controlling viral titers, even before 5 d.p.i. Beta2-microglobulinde cient mice, which lack CD8 C T cells, have delayed clearance of virus from the liver .
cord-312051-2dfc9xjt	2013	
cord-312294-c9e18rai	1994	title: Increased viral titers and enhanced reactivity of antibodies to the spike glycoprotein of murine coronavirus produced by infection at pH 6 Abstract Infection of cell monolayers by murine coronavirus A59 at pH 6 rather than 7 yielded a ten-fold increase in the infectious titer and a remarkable enhancement of the reactivities of monoclonal and polyclonal antibodies against the spike glycoprotein in immunoblotting, immuno-precipitation and enzyme-linked immunosorbent assays. In this study, we show that infection by MHV-A59 in culture medium at pH 6, instead of the usual pH 7, not only increased infectious viral titers, but also greatly enhanced the reactivity of the spike glycoprotein to antibodies. Metabolically labeled virus-infected cell lysates were immunoprecipitated with MHV-specific antibodies ((Y-AS9: hyperimmune serum; 7-10A: anti-S MAb; 653: control ascites fluid) and resolved by SDS-PAGE on a 7-15% gel. Monoclonal antibodies to murine hepatitis virus-4 (strain JHM) define the viral glycoprotein responsible for attachment and cell-cell fusion
cord-313263-epylkxdx	2010	
cord-317169-qlqavi4t	2015	(Saururaceae) and three of its constituent flavonoids (quercetin, quercitrin and rutin) against murine coronavirus and dengue virus (DENV). cordata and flavonoids were assessed using virus neutralization tests against mouse hepatitis virus (MHV) and DENV type 2 (DENV-2). Certain flavonoids exhibited comparatively weaker antiviral activity, notably quercetin which could inhibit both MHV and DENV-2. This was followed by quercitrin which could inhibit DENV-2 but not MHV, whereas rutin did not exert any inhibitory effect on either virus. cordata contribute to the superior antiviral efficacy of the EA fraction which lacked cytotoxicity in vitro and acute toxicity in vivo. cordata exhibited anti-MHV activity at a MIC of 0.98 mg/mL without any apparent cytotoxic effects on CCL9.1 cells. cordata and its flavonoid component, quercetin, could inhibit both MHV and DENV-2 in vitro. Houttuynia cordata extracts and constituents inhibit the infectivity of dengue virus type 2 in vitro
cord-317635-jal2tkra	2005	
cord-320165-1b6sycgv	2020	
cord-324321-y96x8x3h	2003	Furthermore, treatment of cells with chloroquine, which blocks the acidification of endosomes, significantly inhibited the repression of the BNip3 promoter activity induced by the acidic pH-dependent MHV mutant OBLV60, which enters cells via endocytosis, indicating that the down-regulation of BNip3 expression is mediated by fusion between viral envelope and cell membranes during entry. To establish further the specificity of the down-regulation of BNip3 gene expression, we used vesicular stomatitis virus (VSV) in a parallel experiment, because VSV is also an enveloped RNA virus whose infection has a broad host cell range and is mediated by the interaction between the envelope G protein and cell membranes (Riedel et al., 1984) . By extrapolating the data from the mutant virus, we can envision that fusion between viral envelope and plasma/endosomal membranes during wild-type MHV infection is involved in the down-regulation of BNip3 gene expression.
cord-325624-6anybxnk	2009	
cord-329036-4bf8eiix	1994	
cord-330266-uypjqif7	2020	
cord-330907-srb8ac7l	1997	We have previously shown that C12 has two amino acid substitutions relative to wild type virus in the spike protein, Q159L (within a region of S1 shown to bind to viral receptor in anin vitroassay) and H716D (in the proteolytic cleavage recognition site). for the altered pathogenic properties of the mutant viruses, we compared the sequence of the spike (S) genes The murine coronavirus, mouse hepatitis virus strain of wild type and mutant viruses isolated from persistently A59 (MHV-A59), produces both hepatitis and neurologiinfected glial cells cultures. We have shown previously that the spike (S) protein from the supernatant of either the ''''C'''' culture of persisencoded by the C12 mutant has two amino acid substitutently infected glial cells at 1 week (C3), 6 weeks (C5), tions as compared with the wild type S protein.
cord-331267-j9ld7q70	2010	The effects of telbivudine on virus replication and cytokine production were investigated in vitro using MHV‐3‐infected macrophages, and the effects on T‐cell response were investigated in vivo in an MHV‐3‐induced viral hepatitis model. The aims of the present study were to characterize the effects of telbivudine on cytokine profile and T-cell response in vitro and in vivo using a previously characterized mouse model of viral hepatitis induced by the coronavirus mouse hepatitis virus strain 3 (MHV-3) [18, 19] . Cytokine level measurement by PCR and enzyme-linked immunosorbent assay To determine the effect of telbivudine on tumour necrosis factor (TNF)-a and interleukin (IL)-12 cytokine production, one million macrophages from BALB/cJ mice were stimulated with MHV-3 (multiplicity of infection, 2.5) and telbivudine was added at indicated concentrations. In summary, the present study demonstrates that the beneficial effects of telbivudine in MHV-3-induced hepatitis may be mediated by its effect on the immune response rather than the inhibition of viral replication in this animal model.
cord-333473-c1lykari	2016	Also, it has been employed in the study of the replication of large DNA viruses; namely, human cytomegalovirus [17] [18] , Kaposi''s sarcoma-associated herpesvirus [19] , herpes simplex virus 1 [20] , vaccinia virus [21] , and bacteriophage lambda [22] , providing insights into the temporal regulation of gene expression in these viruses and identifying numerous previously unrecognized translated ORFs, including novel protein-coding ORFs and short regulatory uORFs. In this paper, we describe the first analysis of RNA virus replication and gene expression by ribosome profiling (and parallel RNASeq), using MHV as a model system. The latter are calculated on a per gene (rather than per transcript) basis, using RNASeq and RiboSeq reads contained entirely within annotated CDS regions (i.e. excluding 5 0 and 3 0 UTRs and also RPFs accumulating at or near to initiation or termination sites), and, like the virus values, are expressed relative to the mean levels for the cell (due to normalization by library size).
cord-333525-67bbmo4m	2013	In the current study, we analyzed the effects of the Endo charge-rich motif on virion incorporation of MHV S protein through substitutions of the homologous regions from the alphacoronavirus porcine transmissible gastroenteritis virus (TGEV), the betacoronaviruses bovine coronavirus (BCoV) and SARS-CoV, or the gammacoronavirus avian infectious bronchitis virus (IBV). Although the S2 portions of coronavirus S proteins show some degree of conservation, the Tm and Endo domains are highly divergent, with the exception of a conserved cluster of seven hydrophobic residues (WPWYVWL) at the start of Tm. To evaluate the functionality of different C-terminal sequence motifs in the MHV S protein, we constructed two sets of mutants in which the ectodomain of either S protein or HK protein was fused to the Tm and Endo domains from TGEV (an alphacoronavirus), BCoV (a betacoronavirus), SARS-CoV (a betacoronavirus), or IBV (a gammacoronavirus) ( Fig. 2A) . Reverting mutations in TGEV chimeras improved S assembly by eliminating positively charged residues in the endodomain MHV S protein mutants containing the entire carboxy terminus or just the charge-rich motif of TGEV S (Mut-TGEV and Mut-MMT) produced irregular plaques (Figs.
cord-334134-fhie2m3u	2012	
cord-334499-fz7vrnb1	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). 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 immune-mediated, as mice that lack T lymphocytes fail to develop disease despite succumbing to encephalitis with high levels of infectious virus in the CNS. Demyelinating disease induced during MHV-JHM infection is partly immune mediated, as mice lacking the ability to generate T cell responses fail to develop demyelination, despite high viral loads and widespread inflammation in the CNS of infected mice [19, 20] . Further studies involve introduction of other chemoattractants into recombinant MHV-JHM, to evaluate their role in demyelinating disease, cell recruitment, generation of immune responses, and clearance of infectious virus in MHV-JHM-infected mice.
cord-336416-vas0b6dt	1980	ABSTRACT The murine coronavirus JHM induces in weanling rats different types of central nervous diseases ranging from an acute panencephalitis to a late demyelinating encephalomyelitis. Of particular interest are the central nervous system (CNS) diseases associated with this virus group, especially the mouse hepatitis virus strain JHM reveals a distinct neurovirulence for mice and rats (1, 2, 3, 4) . In the present communication the neurovirulence of four murine coronavirus strains (MHV 1, MHV 2, MHV 3 and MHV A59) for rats is compared to the JHM virus. The occurrence and rate of the different types of CNS disease induced by JHM virus is associated with the properties of the virus preparation used as inoculum as summarized in table 2. Reisolated mutants from diseased animals with LDE did not always maintain their temperature sensitivity but were different from revertants by the type of neurovirulence.
cord-337976-c2auspti	1983	A cDNA probe representing the genome of mouse hepatitis virus (MHV) strain A59 (MHV-A59) was used to measure nucleotide sequence homologies among murine and human coronaviruses and the SD and SK coronaviruses isolated by Burks et al. Since SD and SK were isolated by inoculation of multiple sclerosis (MS) central nervous system (CNS) tissue into mice or cultured mouse cells, it is important to determine their relationships to other murine and human coronavirus isolates. Some strains of HCV such as 0C43, are antigenically related to murine coronaviruses such as MHV strain JHM (McIntosh, 1974; Gerdes et al., 1981a, b) and may be grown in the brains of suckling mice (McIntosh et al., 1967) . We have further compared murine and human coronaviruses and SD and SK by using molecular hybridization of virus-specific RNA with cDNA probes. RNA extracted either from brain homogenates of OC43-infected suckling mice or from HRT cells infected with OC43 shows homology with A59 cDNA when assayed by blot hybridization.
cord-338165-mgrf1odm	2012	Virus titers were determined in the liver tissue of MHV-3 infected C3H/HeJ mice at various time points by standard plaque assay as described before [32] . Cell suspensions of processed spleens from C3H/HeJ mice on days 4, 10, 15, 20, and 30 post MHV-3 infection were stained with PEcy5.5-anti-CD3, FITC-anti-CD4, FITC-anti-CD8, APC-anti-CD25, APC-anti-TCRb, PE-anti-TCRcd, PE-anti-CD28/PE-anti-CD30, PE-anti-CD44, PE-anti-CD95L, PEanti-CD95 monoclonal antibodies or isotype control Ab (eBioscience, San Diego, CA, USA). Adoptive transfer of DN T cells from MHV-3 infected C3H/HeJ mice increased the survival rate and improved liver histology of recipient mice infected by the same virus strain but had little impact on the virus titer of liver tissue Fig. 3A showed that no significant difference in virus titer of liver tissue was observed among DN T cells group, splenocytes group, DNT-depleted splenocytes group and PBS control group.
cord-338307-vfutmwxq	1983	The pattern of three major structural proteins and their organization in the virion as shown for MHV-A59 in Fig. 3 is generally applicable to most other species of coronaviruses (reviewed in detail by Siddell et al., 1982) . (1981b) , using a cDNA probe prepared against the mRNA of MHV-A59 which coded for the nucleocapsid protein, obtained evidence of 7 0 4 0 % homology by analysis of hybridization kinetics of viral RNA from cells infected with MHV-1, -3, -S, and -JHM. t 1980) demonstrated that different size classes of poly(A)containing intracellular MHV-JHM RNAs fractionated on sucroseformamide gradients directed the synthesis of different structural proteins in cell-free translation systems. The studies showed that, like avian coronaviruses, for the murine coronavirus MHV-A59, the oligonucleotides of each of the subgenomic RNAs were included within the next larger species, starting from the 3'' end of the genome.
cord-341278-klv9jdm8	1991	
cord-341342-kyavg4vu	1992	The RNase sensitivity of the ability of N protein to migrate into nondenaturing gels was not due to possible contaminating protease activity in the RNase preparations, since the same RNase-treated samples of translated N 148 P.S. Masters protein showed no degradation when analyzed by SDS-PAGE (Fig. 3, lanes an, lower) . Since the N mRNAs synthesized from the transcription vectors shown in Fig. 1 contained this portion of the leader sequence, it seemed possible that the N-RNA complex observed in nondenaturing PAGE was that of translated N protein binding to its own mRNA. That the observed complex was not due to translated N protein binding to the leader portion of its own mRNA was also supported by the observation that full-length N protein translated from a construct in which the MHV leader sequence was entirely deleted was also able to migrate into nondenaturing gels (data not shown).
cord-342151-1e6x589e	2008	Erythrocytes from twelve mammalian and avian sources in ten different buffers at three incubation temperatures could not be hemagglutinated with murine hepatitis virus (MHV) strains 3, A59, or S grown on DBT cells. Viral antigen preparation in the absence of fetal calf serum, partial virus purification, or various concentrations of red blood cells still failed to yield detectable hemagglutinating activity. For use as antigen for hemag glutination assays, MHV-3 was grown in the absence of fetal calf serum (FCS) and har vested from clarified medium by precipita tion with 10% (w/v) polyethylene glycol in 0.5 M NaCl. After centrifugation at 10,000 g for 30 min, the pellet was resuspended and dialyzed against TMEN buffer: 50 mM Tris-HCi (pH 6.2), 0.1 M NaCl, 1 m M EDTA. Control hemagglutinating antigens were either rabbit enteric coronavirus (titer 1/64 with rabbit red blood cells ) or pneumonia virus of mice (titer 1/320 with CDI mouse erythrocytes).
cord-343221-e29of29o	2017	Specifically, we show that genetically engineered mutants of mouse hepatitis virus (MHV) and human coronavirus 229E (HCoV-229E), respectively, that encode an EndoU active-site substitution known to abolish nucleolytic activity, were severely attenuated and elicited an immediate and simultaneous activation of host cell dsRNA sensors. The severe attenuation of MHV H277A and HCoV-229E H250A replication in wild-type murine and human macrophages, respectively, precluded the use of primary macrophages to assess the sensitivity of EndoU-deficient coronaviruses to IFN-I pre-treatment. Our data suggest that direct restriction of replication of EndoU-deficient coronaviruses is mediated by RNase L-mediated RNA degradation and inhibition of host cell translation through activation of PKR. Notably, also in this case Xrn1 is required to further process the de-capped mRNAs. Our data show that early during coronavirus infection the EndoU activity conceptually fulfils the same task, namely the removal of dsRNA that would otherwise trigger host cell dsRNA responses, such as IFN-β expression and activation of PKR and OAS/RNase L.
cord-345630-bam3pa70	1991	authors: Lee, Han-Jung; Shieh, Chien-Kou; Gorbalenya, Alexander E.; Koonin, Eugene V.; La Monica, Nicola; Tuler, Jeremy; Bagdzhadzhyan, Anush; Lai, Michael M.C. title: The complete sequence (22 kilobases) of murine coronavirus gene 1 encoding the putative proteases and RNA polymerase Third, the 3''-half of the gene 1 sequences of IBV and MHV-A59 contains the sequence motifs for RNA polymerase and helicase, which are the activities expected to be involved in RNA synthesis Gorbalenya et a/., 198913; Bredenbeek et a/., 1990) . The alignment of amino acids in ORF la of MHV-JHM and IBV showed that there are four possible stretches of moderate homology which are separated by highly diverged sequences (Fig. 8) . Although ORF la is highly diverged between MHV-JHM and IBV, common functional domains could be identified in this ORF of both viruses by detailed amino acid sequence analysis (see Materials and Methods) (Fig. 9 ).
cord-348746-yaf61cmx	2008	F eline infectious peritonitis (FIP) is a fatal, immune-mediated disease produced as a result of infection of macrophages by mutant feline coronavirus strains (FIPVs). In acute MHV-A59 infection in CD8Ï© T-cell deficient mice, periventricular encephalitis occurs with lymphocytic infiltration into the choroid plexus, ependyma, and subependymal brain tissue. Depending on mouse strain and immunological status, MHV-JHM produces meningeal inflammation associated with T-cells and macrophages and demyelination but relatively little disease in axons. If mice are pretreated with passive infusions of antibodies or T-cells or if they receive neuroattenuated MHV strains, they develop chronic, but not fatal, disease after MHV-JHM infection. 62, 63 Immunocompetent C57BL/6 mice clear MHV-JHM virus from the brain but develop severe immune-mediated demyelination and paralysis. Two related strains of feline infectious peritonitis virus isolated from immunocompromised cats infected with a feline enteric coronavirus
cord-349135-it5ahzj3	2006	
cord-351934-g7tgo5cn	2006	authors: Deming, Damon J.; Graham, Rachel L.; Denison, Mark R.; Baric, Ralph S. Through use of an efficient MHV-A59 reverse genetics system, 6 we ablated each of the M pro cleavage sites associated with the nsp7-nsp10 cassette, and evaluated whether the mutated genome was capable of supporting a viable virus, and if so, characterized the M pro processing of the mutated protein, transcription function, and in vitro growth fitness. Surprisingly, the recovered virus did not revert to wild-type sequence at the nsp9/nsp10 M pro cleavage site, indicating that an as of yet unidentified mutation(s) has compensated for the virus''s inability to properly process the nsp9-nsp10 precursor protein. Serial passage of this virus restored wild-type replication but did so without reverting the mutated cleavage site or the ability to process the nsp9-nsp10 protein. The data demonstrate that with the exception of cleavage between the nsp9 and nsp10 proteins, M pro processing of the nsp7-nsp10 cassette is essential in coronavirus RNA transcription and replication.
cord-351964-hduv0ur4	1987	
cord-352379-q5inrxcm	2003	Nevertheless, the lack of a firm association of coronaviruses with any serious human illnesses had dampened the public''s interest in this virus family until the sudden emergence of the SARS coronavirus [24, 41, 62] , which caused the first new infectious disease of this millennium. In the SARS virus genome, the organization of gene la-lb, which accounts for more than two-thirds of the viral RNA, is very similar to that of the murine coronavirus MHV, except that it contains only one papain-like protease (PLpro-2) ( fig. Based on the predicted cleavage site specificity, the SARS virus gene la-lb is likely processed into thirteen final protein products. However, the published sequence analysis indicated that the entire SARS virus RNA resembled that of group II viruses; no evidence of recombination was noted [55, 66] . Comparative full-length genome sequence analysis of 14 SARS coronavirus isolates and common mutations associated with putative origins of infection
cord-353190-7qcoxl81	2012	This chapter covers infections of mice with the following viruses: herpesviruses, mousepox virus, murine adenoviruses, polyomaviruses, parvoviruses, lactate dehydrogenase-elevating virus, lymphocytic choriomeningitis virus, mammalian orthoreovirus serotype 3, murine hepatitis virus, murine norovirus, murine pneumonia virus, murine rotavirus, Sendai virus, and Theiler''s murine encephalomyelitis virus. These results are very difficult to summarize because the outcome of experimental infection in laboratory mice depends on various factors such as mouse strain and age, virus strain and passage history [26] , virus dose and route of inoculation [24] . Experimental infection of laboratory mice with MHV-68 is a frequently used model system for the study of human gammaherpesvirus pathogenesis, e.g. of Kaposi''s sarcoma-associated herpesvirus or Epstein-Barr virus (EBV) [62, 63] which are members of the same subfamily. Early descriptions of naturally occurring disease may have been complicated by concurrent infections such as MHV (murine hepatitis virus) or murine rotavirus A (MuRV-A)/epizootic diarrhoea of infant mice (EDIM) virus that contributed to the severity of the lesions especially in liver, pancreas, CNS and intestine.
cord-354726-b9xvycyk	1995	
cord-356013-pl3tmky8	2005	
cord-356115-vblgotjn	2005	The coronavirus replicase-transcriptase complex is an assembly of viral and cellular proteins that mediate the synthesis of genome and subgenome-sized mRNAs in the virus-infected cell. Here, we report a genetic and functional analysis of 19 temperature-sensitive (ts) mutants of Murine hepatitis virus MHV-A59 that are unable to synthesize viral RNA when the infection is initiated and maintained at the non-permissive temperature. We focused our phenotypic analysis on the eight MHV-A59 ts mutants that had been genotyped and began by measuring ''''total'''' viral RNA synthesis in infected cells prior to and following shift from the permissive to the non-permissive temperature. This phenotype resembled that seen with MHV-A59-infected cells treated with CH, and we conclude that the complementation group I mutants are defective in their ability to form active replicase-transcriptase complexes at 40 8C but retain the positive-strand synthesis activity of the complexes formed at the permissive temperature.