RESUMO
Theiler's murine encephalomyelitis virus (TMEV) belongs the family Picornaviridae. TMEV not only replicates in the gastrointestinal tract but also spreads to the central nervous system (CNS) either by a hematogenous or a neural pathway during natural infection. The DA strain of TMEV infects neurons during the acute phase, and glial cells and macrophages during the chronic phase, leading to a demyelinating disease similar to multiple sclerosis. Different virus-host receptor interactions in the peripheral and the neuronal cells could explain the pathways of viral spread from the peripheral to the CNS and neurons to glial cells. However, the receptor for TMEV remains unknown. P0 protein, a 28-31 kD glycoprotein, belongs to the immunoglobulin superfamily and constitutes 50% of the total myelin protein in the peripheral nerve. Other picornaviruses use members of the immunoglobulin superfamily as receptors. Thus we hypothesized P0 protein could act as a receptor for TMEV. In a virus overlay assay, radiolabeled TMEV bound to a 28-30 kD protein from the peripheral nerve of wild-type C57BL/6, but no binding was found in the peripheral nerve from P0-knockout mice. TMEV replicated fourfold higher in P0-transfected BW5147.G.1.4 cells than in mock-transfected cells. The increase in virus replication in the P0-transfected cell line was blocked by preincubation of the cells with anti-P0 antibody. A virus binding study showed that TMEV bound to P0-transfected cells but not to mock-transfected cells. The use of the P0 protein in Schwann cells as a receptor may be one mechanism by which TMEV spreads from the gastrointestinal tract to the CNS.
Assuntos
Infecções por Cardiovirus/metabolismo , Proteína P0 da Mielina/metabolismo , Nervo Isquiático/virologia , Theilovirus , Sequência de Aminoácidos , Animais , Anticorpos/farmacologia , Capsídeo/genética , Proteínas do Capsídeo , Células Cultivadas , Cricetinae , Expressão Gênica/fisiologia , Rim/citologia , Camundongos , Camundongos Knockout , Dados de Sequência Molecular , Proteína P0 da Mielina/genética , Proteína P0 da Mielina/imunologia , Nervo Isquiático/citologia , TransfecçãoRESUMO
The Gene Expression Database (GXD) is a community resource of gene expression information for the laboratory mouse. By combining the different types of expression data, GXD aims to provide increasingly complete information about the expression profiles of genes in different mouse strains and mutants, thus enabling valuable insights into the molecular networks that underlie normal development and disease. GXD is integrated with the Mouse Genome Database (MGD). Extensive interconnections with sequence databases and with databases from other species, and the development and use of shared controlled vocabularies extend GXD's utility for the analysis of gene expression information. GXD is accessible through the Mouse Genome Informatics web site at http://www.informatics.jax.org/ or directly at http://www.informatics.jax.org/menus/expression_menu. shtml.
Assuntos
Bases de Dados Factuais , Perfilação da Expressão Gênica , Camundongos/genética , Animais , Serviços de Informação , InternetRESUMO
Theiler's murine encephalomyelitis viruses are picornaviruses that can infect the central nervous system. The DA strain produces an acute polioencephalomyelitis followed by a chronic demyelinating disease in its natural host, the mouse. The ability of DA virus to induce a demyelinating disease renders this virus infection a model for human demyelinating diseases such as multiple sclerosis. Here we describe the generation and characterization of DA virus mutants that contain specific mutations in the viral capsid protein VP1 at sites believed to be important contact regions for the cellular receptor(s). A mutant virus with a threonine-to-aspartate (T81D) substitution in VP1 loop I adjacent to the putative virus receptor binding site exhibited a large-plaque phenotype but had a slower replication cycle in vitro. When this mutant virus was injected into susceptible mice, an altered tropism was seen during the acute stage of the disease and the chronic demyelinating disease was not produced. A virus with a threonine-to-valine substitution (T81V) did not cause any changes in the pattern or extent of disease seen in mice, whereas a virus with a tryptophan substitution at this position (T81W) produced a similar acute disease but was attenuated for the development of the chronic disease. A change in amino acids in a hydrophobic patch located in the wall of the pit, VP1 position 91, to a hydrophilic threonine (V91T) resulted in a profound attenuation of the acute and chronic disease without persistence of virus. This report illustrates the importance of the loop I of VP1 and a site in the wall of the pit in pathogenesis and that amino acid substitutions at these sites result in altered virus-host interactions.
Assuntos
Capsídeo/genética , Infecções por Cardiovirus/virologia , Theilovirus/genética , Animais , Proteínas do Capsídeo , Linhagem Celular , Humanos , Imuno-Histoquímica , Camundongos , Modelos Moleculares , Mutação , Theilovirus/patogenicidade , Virulência/genética , Replicação Viral/genéticaRESUMO
Theiler's murine encephalomyelitis viruses, which are murine picornaviruses, can cause central nervous system inflammatory disease. To study the role of loop II in capsid protein VP1, two mutant viruses of strain DA in which DA loop II amino acids were replaced with strain GDVII amino acids were constructed. Infection of mice with the two mutant viruses led to dramatically different patterns of disease.
Assuntos
Capsídeo/genética , Capsídeo/fisiologia , Doenças Desmielinizantes/patologia , Poliomielite/patologia , Theilovirus/fisiologia , Animais , Encéfalo/virologia , Capsídeo/química , Proteínas do Capsídeo , Linhagem Celular , Cricetinae , Camundongos , Mutagênese Sítio-Dirigida , Poliomielite/virologia , Conformação Proteica , Medula Espinal/patologia , Theilovirus/genética , Theilovirus/patogenicidade , Virulência , Latência ViralRESUMO
The etiology of hydrocephalus is never established in the majority of clinical cases, while various agents, nutritional deficiencies, and genetic factors have been shown to play a role. Viral infection has been recognized as one of the causative factors in the development of hydrocephalus. The wild-type DA strain of Theiler's murine encephalomyelitis virus (TMEV), which belongs to the family Picornaviridae, causes a chronic demyelinating disease in mice with viral persistence that resembles multiple sclerosis. We found that a DA virus variant, hydrocephalus 101 virus (H101 virus), caused hydrocephalus in mice, a condition previously never described for TMEV. To clarify the relationship between DA virus infection and hydrocephalus, we compared H101 virus and wild-type DA virus infection in mice. Using immunohistochemistry and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL), we found that during the acute phase of infection, H101 virus caused macrocephaly and meningitis with the presence of apoptosis, while parenchymal involvement was not evident. In contrast, wild-type DA virus caused an acute polioencephalomyelitis with parenchymal infection and apoptosis. During the chronic phase, H101 virus infection caused communicating hydrocephalus without viral persistence. No demyelination and little or no anti-TMEV antibodies were observed in H101 virus-infected mice. Sequence analysis revealed that H101 virus had mutations in the 5'UTR and capsid protein coding region. Characterization of this new hydrocephalus model gives insight into the possible viral involvement in human hydrocephalus cases of obscure etiology.
Assuntos
Variação Genética/genética , Hidrocefalia/virologia , Poliomielite/complicações , Theilovirus/genética , Animais , Anticorpos Antivirais/análise , Sequência de Bases , Sistema Nervoso Central/virologia , Feminino , Predisposição Genética para Doença , Genoma Viral , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos , Poliomielite/patologia , Poliomielite/fisiopatologia , Theilovirus/imunologiaRESUMO
The DA strain of Theiler's murine encephalomyelitis virus (TMEV) is a picornavirus which infects the murine central nervous system (CNS). During the acute stage versus the chronic stage, different cell-types are infected. To clarify the shift seen during persistent infection, we have screened various cell lines for their ability to support TMEV infection and used virus binding assays to determine if there was a correlation between permissiveness and virus binding. Cell lines of different origins were able to support virus infection to varying degrees. Infectious center assays and viral binding assay demonstrated that permissiveness to virus infection and the ability of virus to attach to the cell surface did not always correlate.