Recovery of a persistent Canine distemper virus expressing the enhanced green fluorescent protein from cloned cDNA

Plattet, Philippe; Zweifel, C.; Wiederkehr, Christa; Belloy, Luc; Cherpillod, Pascal; Zurbriggen, Andreas; Wittek, Riccardo

doi:10.1016/j.virusres.2004.01.002

Cited by 31 publications

(24 citation statements)

References 45 publications

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“…Recombinant MV wild-type strain IC323-eGFP [rMV-IC323-eGFP (Hashimoto et al, 2002)] was propagated using Vero-hSLAM. CDV strain Onderstepoort large plaque was propagated in Vero cells and recombinant CDV wild-type strain A75/17, expressing tomato red [rCDV-A75/17red, (Plattet et al, 2004)] was propagated in Vero-dogSLAM cells.…”

Section: Methodsmentioning

confidence: 99%

N-(3-Cyanophenyl)-2-phenylacetamide, an effective inhibitor of morbillivirus-induced membrane fusion with low cytotoxicity

Singethan

Hiltensperger

Kendl

et al. 2010

Journal of General Virology

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Based on the structural similarity of viral fusion proteins within the family Paramyxoviridae, we tested recently described and newly synthesized acetanilide derivatives for their capacity to inhibit measles virus (MV)-, canine distemper virus (CDV)-and Nipah virus (NiV)-induced membrane fusion. We found that N-(3-cyanophenyl)-2-phenylacetamide (compound 1) has a high capacity to inhibit MV-and CDV-induced (IC 50 53 mM), but not NiV-induced, membrane fusion. This compound is of outstanding interest because it can be easily synthesized and its cytotoxicity is low [50 % cytotoxic concentration (CC 50 )¢300 mM], leading to a CC 50 /IC 50 ratio of approximately 100. In addition, primary human peripheral blood lymphocytes and primary dog brain cell cultures (DBC) also tolerate high concentrations of compound 1. Infection of human PBMC with recombinant wild-type MV is inhibited by an IC 50 of approximately 20 mM. The cell-tocell spread of recombinant wild-type CDV in persistently infected DBC can be nearly completely inhibited by compound 1 at 50 mM, indicating that the virus spread between brain cells is dependent on the activity of the viral fusion protein. Our findings demonstrate that this compound is a most applicable inhibitor of morbillivirus-induced membrane fusion in tissue culture experiments including highly sensitive primary cells. INTRODUCTIONThe genus of closely related viruses Morbillivirus, contains measles virus (MV) and canine distemper virus (CDV), which cause devastating diseases in their hosts. MV, one of the most contagious aerosol-transmitted viruses, still causes more than 100 000 deaths each year. After entering the upper respiratory tract, MV exhibits a pronounced tropism for mono-and lymphocytic cells and soon viral replication is detected in draining lymph nodes (Pfeuffer et al., 2003; de Swart et al., 2007;Ferreira et al., 2010). During measles, patients develop a pronounced leukopenia that may be due to enhanced adhesion of lymphocytes in secondary lymphoid organs (Nanan et al., 1999; Dittmar et al., 2008). In immunocompetent patients, MV infection is usually cleared by the virus-specific immune response, while the general immune response to other antigens is suppressed for several weeks after the rash (reviewed by ). Following replication in lymphoid tissues, virus spreads to various organs and can be detected in the skin, the gastrointestinal tract, the lungs and the eyes. It may also enter the brain (reviewed by Ludlow et al., 2009). CNS complications are acute post-infectious measles encephalitis and, based on a persistent infection, subacute sclerosing pan-encephalitis (SSPE), which is always lethal. SSPE does not occur after measles vaccination (Duclos & Ward, 1998).CDV causes systemic infections similar to but distinct from human measles in carnivores such as canines, felids, ferrets, raccoons and seals, with lethality rates, depending on the host, of up to 100 % (Appel et al., 1972;Harder & Osterhaus, 1997;von Messling et al., 2003). In a high percentage of animals the CNS is inf...

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Section: Methodsmentioning

confidence: 99%

N-(3-Cyanophenyl)-2-phenylacetamide, an effective inhibitor of morbillivirus-induced membrane fusion with low cytotoxicity

Singethan

Hiltensperger

Kendl

et al. 2010

Journal of General Virology

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“…Recombinant viruses were generated as described previously (26,43). Briefly, BsrT7 cells in 6-well plates were cotransfected with 2.5 g of full-length cDNA of the relevant clones together with the expression vectors encoding the N, P, and L proteins (0.2, 0.2, and 0.05 g, respectively).…”

Section: Vol 85 2011 Insights Into CDV Multiple-receptor Usage 11243mentioning

confidence: 99%

“…To gain further insight into CDV-mediated multiple-receptor usage, we used the virulent CDV strain A75/17, which was isolated directly from lymphoid tissues of a naturally CDV infected dog and, importantly, was never adapted to grow in any cell lines. This virus, for which efficient reverse genetics is available (26,27), grows well in several primary canine cell cultures, where it closely reproduces the selective noncytolytic cell-to-cell spread seen in vivo. In this study, we propose that CDV-H may interact with receptors similar to those for MeV.…”

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confidence: 99%

Canine Distemper Virus Infects Canine Keratinocytes and Immune Cells by Using Overlapping and Distinct Regions Located on One Side of the Attachment Protein

Langedijk

Janda

Origgi

et al. 2011

J Virol

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The morbilliviruses measles virus (MeV) and canine distemper virus (CDV) both rely on two surface glycoproteins, the attachment (H) and fusion proteins, to promote fusion activity for viral cell entry. Growing evidence suggests that morbilliviruses infect multiple cell types by binding to distinct host cell surface receptors. Currently, the only known in vivo receptor used by morbilliviruses is CD150/SLAM, a molecule expressed in certain immune cells. Here we investigated the usage of multiple receptors by the highly virulent and demyelinating CDV strain A75/17. We based our study on the assumption that CDV-H may interact with receptors similar to those for MeV, and we conducted systematic alanine-scanning mutagenesis on CDV-H throughout one side of the ␤-propeller documented in MeV-H to contain multiple receptor-binding sites. Functional and biochemical assays performed with SLAM-expressing cells and primary canine epithelial keratinocytes identified 11 residues mutation of which selectively abrogated fusion in keratinocytes. Among these, four were identical to amino acids identified in MeV-H as residues contacting a putative receptor expressed in polarized epithelial cells. Strikingly, when mapped on a CDV-H structural model, all residues clustered in or around a recessed groove located on one side of CDV-H. In contrast, reported CDV-H mutants with SLAM-dependent fusion deficiencies were characterized by additional impairments to the promotion of fusion in keratinocytes. Furthermore, upon transfer of residues that selectively impaired fusion induction in keratinocytes into the CDV-H of the vaccine strain, fusion remained largely unaltered. Taken together, our results suggest that a restricted region on one side of CDV-H contains distinct and overlapping sites that control functional interaction with multiple receptors.

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“…After digestion with NotI and PmeI, the M, F, and H genes of CDV A75/17 and a red fluorescence marker gene (tD-tomato; kindly offered by D. Garcin, University of Geneva, Switzerland) (23) placed between the first two genes were cloned into the shuttle vectors, which contained the mutated P genes, using T4 DNA ligase (New England BioLabs). To rescue the recombinant viruses we proceeded as previously described (33). Briefly, Bsr-T 7 cells were cotransfected with plasmids containing the relevant full-length cDNA and helper plasmids coding for the N, P, and L proteins of CDV.…”

Section: Cells and Virusesmentioning

confidence: 99%

“…84, 2010 MECHANISMS OF CDV-V AS IFN ANTAGONIST 6329 pCI-HA-VCT, and pCI-HA-Vwt Y110D. The green fluorescent protein (GFP) gene was amplified by PCR technology from a plasmid pgA75/17-V cDNA clone (33) and subsequently digested and ligated into the pCI-cleaved plasmid, thus generating pCI-GFP. Luciferase assay and transient transfections.…”

Section: Cells and Virusesmentioning

confidence: 99%

Two Domains of the V Protein of Virulent Canine Distemper Virus Selectively Inhibit STAT1 and STAT2 Nuclear Import

Röthlisberger

Wiener

Schweizer

et al. 2010

J Virol

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Canine distemper virus (CDV) causes in dogs a severe systemic infection, with a high frequency of demyelinating encephalitis. Among the six genes transcribed by CDV, the P gene encodes the polymerase cofactor protein (P) as well as two additional nonstructural proteins, C and V; of these V was shown to act as a virulence factor. We investigated the molecular mechanisms by which the P gene products of the neurovirulent CDV A75/17 strain disrupt type I interferon ( Virulent canine distemper virus (CDV) causes a severe systemic infection in dogs that is characterized by high fever, diarrhea, and pneumonia. Large-scale immunosuppression is a hallmark of infection, and some virus strains additionally invade the central nervous system to cause chronic demyelinating encephalitis. The molecular mechanisms differentiating virulent from attenuated strains are poorly understood. However, the fact that dogs can be protected from infection with virulent CDV by vaccination with attenuated strains suggests that reliable induction of adaptive immunity is possible, provided that the critical early stage of infection is successfully mastered by the host. During the early stage of infection, host defense depends on the innate immune system, which is also responsible for generating signals that activate the adaptive immune response (27). The interferons of type I (IFN-I, e.g., IFN-␣/␤) are a critical element of the innate immune defense against viruses (13,36,41). Virtually all nucleated cells are capable of sensing viral infection by receptors such as Rig-I, MDA-5, or Toll-like receptor-3 (16). Activation of these receptors initiates a signal cascade that results in transcription, translation, and release from the cells of IFN-␣/␤. This part of the IFN defense is referred to as the induction stage. IFN action is initiated by the binding of IFN to type I IFN receptors that activates the receptor-associated tyrosine kinases JAK1 and Tyk2, which, in turn, phosphorylate the signal transducers and activators of transcription (STATs) (21, 41). Subsequently, the activated STAT1 and STAT2 together with IFN regulatory factor 9 (IRF9) form a complex, the IFN-stimulated gene factor 3 (ISGF3), which, once translocated to the nucleus, binds the IFN-stimulated response element (ISRE) sequence (39,45). This initiates the expression of well over 100 proteins which are responsible for the antiviral effect of IFN (36). In recent years, gene products targeting specific steps of IFN induction or action have been found in virtually all viruses studied, indicating the crucial role of IFN evasion in any successful interaction of viruses with their hosts.CDV, a Morbillivirus of the Paramyxoviridae, contains a nonsegmented, single-stranded, negative-sense RNA genome. The genome consists of six genes expressing the structural nucleocapsid (N), matrix (M), fusion (F), hemagglutinin (H), and large (L) proteins and the phospho-protein (P) (20). The P and the L proteins together form the RNA polymerase. In addition to the P protein, the nonstructural C and V p...

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Recovery of a persistent Canine distemper virus expressing the enhanced green fluorescent protein from cloned cDNA

Cited by 31 publications

References 45 publications

N-(3-Cyanophenyl)-2-phenylacetamide, an effective inhibitor of morbillivirus-induced membrane fusion with low cytotoxicity

N-(3-Cyanophenyl)-2-phenylacetamide, an effective inhibitor of morbillivirus-induced membrane fusion with low cytotoxicity

Canine Distemper Virus Infects Canine Keratinocytes and Immune Cells by Using Overlapping and Distinct Regions Located on One Side of the Attachment Protein

Two Domains of the V Protein of Virulent Canine Distemper Virus Selectively Inhibit STAT1 and STAT2 Nuclear Import

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