scholarly journals Viral Pathogenesis, Recombinant Vaccines, and Oncolytic Virotherapy: Applications of the Canine Distemper Virus Reverse Genetics System

Viruses ◽  
2020 ◽  
Vol 12 (3) ◽  
pp. 339 ◽  
Author(s):  
Jianjun Zhao ◽  
Yanrong Ren ◽  
Jie Chen ◽  
Jiasan Zheng ◽  
Dongbo Sun
Keyword(s):  
Canine Distemper Virus ◽  
Reverse Genetics ◽  
Tumor Regression ◽  
Genetic System ◽  
Viral Pathogenesis ◽  
Oncolytic Viruses ◽  
Reverse Genetic System ◽  
Canine Distemper ◽  
Host Tropism ◽  
Signaling Lymphocytic Activation Molecule

Canine distemper virus (CDV) is a highly contagious pathogen transmissible to a broad range of terrestrial and aquatic carnivores. Despite the availability of attenuated vaccines against CDV, the virus remains responsible for outbreaks of canine distemper (CD) with significant morbidity and mortality in domesticated and wild carnivores worldwide. CDV uses the signaling lymphocytic activation molecule (SLAM, or CD150) and nectin-4 (PVRL4) as entry receptors, well-known tumor-associated markers for several lymphadenomas and adenocarcinomas, which are also responsible for the lysis of tumor cells and apparent tumor regression. Thus, CDV vaccine strains have emerged as a promising platform of oncolytic viruses for use in animal cancer therapy. Recent advances have revealed that use of the CDV reverse genetic system (RGS) has helped increase the understanding of viral pathogenesis and explore the development of recombinant CDV vaccines. In addition, genetic engineering of CDV based on RGS approaches also has the potential of enhancing oncolytic activity and selectively targeting tumors. Here, we reviewed the host tropism and pathogenesis of CDV, and current development of recombinant CDV-based vaccines as well as their use as oncolytic viruses against cancers.

scholarly journals The Hemagglutinin of Canine Distemper Virus Determines Tropism and Cytopathogenicity

2001 ◽  
Vol 75 (14) ◽  
pp. 6418-6427 ◽  
Author(s):  
Veronika von Messling ◽  
Gert Zimmer ◽  
Georg Herrler ◽  
Ludwig Haas ◽  
Roberto Cattaneo
Keyword(s):  
Vaccine Strain ◽  
Measles Virus ◽  
Canine Distemper Virus ◽  
Syncytium Formation ◽  
Genetic System ◽  
Reverse Genetic System ◽  
Canine Distemper ◽  
Large Plaque ◽  
Recombinant Viruses ◽  
H Protein

ABSTRACT Canine distemper virus (CDV) and measles virus (MV) cause severe illnesses in their respective hosts. The viruses display a characteristic cytopathic effect by forming syncytia in susceptible cells. For CDV, the proficiency of syncytium formation varies among different strains and correlates with the degree of viral attenuation. In this study, we examined the determinants for the differential fusogenicity of the wild-type CDV isolate 5804Han89 (CDV5804), the small- and large-plaque-forming variants of the CDV vaccine strain Onderstepoort (CDVOS and CDVOL, respectively), and the MV vaccine strain Edmonston B (MVEdm). The cotransfection of different combinations of fusion (F) and hemagglutinin (H) genes in Vero cells indicated that the H protein is the main determinant of fusion efficiency. To verify the significance of this observation in the viral context, a reverse genetic system to generate recombinant CDVs was established. This system is based on a plasmid containing the full-length antigenomic sequence of CDVOS. The coding regions of the H proteins of all CDV strains and MVEdm were introduced into the CDV and MV genetic backgrounds, and recombinant viruses rCDV-H5804, rCDV-HOL, rCDV-HEdm, rMV-H5804, rMV-HOL, and rMV-HOS were recovered. Thus, the H proteins of the two morbilliviruses are interchangeable and fully functional in a heterologous complex. This is in contrast with the glycoproteins of other members of the familyParamyxoviridae, which do not function efficiently with heterologous partners. The fusogenicity, growth characteristics, and tropism of the recombinant viruses were examined and compared with those of the parental strains. All these characteristics were found to be predominantly mediated by the H protein regardless of the viral backbone used.

scholarly journals Intratumoral Canine Distemper Virus Infection Inhibits Tumor Growth by Modulation of the Tumor Microenvironment in a Murine Xenograft Model of Canine Histiocytic Sarcoma

2021 ◽  
Vol 22 (7) ◽  
pp. 3578
Author(s):  
Federico Armando ◽  
Adnan Fayyad ◽  
Stefanie Arms ◽  
Yvonne Barthel ◽  
Dirk Schaudien ◽  
...  
Keyword(s):  
Tumor Microenvironment ◽  
Virus Infection ◽  
Tumor Growth ◽  
Canine Distemper Virus ◽  
Xenograft Model ◽  
Histiocytic Sarcoma ◽  
Oncolytic Viruses ◽  
Canine Distemper ◽  
Murine Xenograft Model ◽  
The Impact

Histiocytic sarcomas refer to highly aggressive tumors with a poor prognosis that respond poorly to conventional treatment approaches. Oncolytic viruses, which have gained significant traction as a cancer therapy in recent decades, represent a promising option for treating histiocytic sarcomas through their replication and/or by modulating the tumor microenvironment. The live attenuated canine distemper virus (CDV) vaccine strain Onderstepoort represents an attractive candidate for oncolytic viral therapy. In the present study, oncolytic virotherapy with CDV was used to investigate the impact of this virus infection on tumor cell growth through direct oncolytic effects or by virus-mediated modulation of the tumor microenvironment with special emphasis on angiogenesis, expression of selected MMPs and TIMP-1 and tumor-associated macrophages in a murine xenograft model of canine histiocytic sarcoma. Treatment of mice with xenotransplanted canine histiocytic sarcomas using CDV induced overt retardation in tumor progression accompanied by necrosis of neoplastic cells, increased numbers of intratumoral macrophages, reduced angiogenesis and modulation of the expression of MMPs and TIMP-1. The present data suggest that CDV inhibits tumor growth in a multifactorial way, including direct cell lysis and reduction of angiogenesis and modulation of MMPs and their inhibitor TIMP-1, providing further support for the concept of its role in oncolytic therapies.

scholarly journals Probing Morbillivirus Antisera Neutralization Using Functional Chimerism between Measles Virus and Canine Distemper Virus Envelope Glycoproteins

Viruses ◽  
2019 ◽  
Vol 11 (8) ◽  
pp. 688 ◽  
Author(s):  
Miguel Angel Muñoz-Alía ◽  
Stephen J. Russell
Keyword(s):  
Neutralizing Antibodies ◽  
Measles Virus ◽  
Canine Distemper Virus ◽  
Reverse Genetics ◽  
Canine Distemper ◽  
Virus Envelope ◽  
Live Virus ◽  
Virus Challenge ◽  
Virus Attachment ◽  
Globular Head

Measles virus (MeV) is monotypic. Live virus challenge provokes a broadly protective humoral immune response that neutralizes all known measles genotypes. The two surface glycoproteins, H and F, mediate virus attachment and entry, respectively, and neutralizing antibodies to H are considered the main correlate of protection. Herein, we made improvements to the MeV reverse genetics system and generated a panel of recombinant MeVs in which the globular head domain or stalk region of the H glycoprotein or the entire F protein, or both, were substituted with the corresponding protein domains from canine distemper virus (CDV), a closely related morbillivirus that resists neutralization by measles-immune sera. The viruses were tested for sensitivity to human or guinea pig neutralizing anti-MeV antisera and to ferret anti-CDV antisera. Virus neutralization was mediated by antibodies to both H and F proteins, with H being immunodominant in the case of MeV and F being so in the case of CDV. Additionally, the globular head domains of both MeV and CDV H proteins were immunodominant over their stalk regions. These data shed further light on the factors constraining the evolution of new morbillivirus serotypes.

scholarly journals Recovery of NanoLuc Luciferase-Tagged Canine Distemper Virus for Facilitating Rapid Screening of Antivirals in vitro

2020 ◽  
Vol 7 ◽  
Author(s):  
Fuxiao Liu ◽  
Qianqian Wang ◽  
Yilan Huang ◽  
Ning Wang ◽  
Youming Zhang ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Canine Distemper Virus ◽  
Reverse Genetics ◽  
Rapid Screening ◽  
Canine Distemper ◽  
Virus Propagation ◽  
The Family ◽  
Conventional Methods

Canine distemper virus (CDV), belonging to the genus Morbillivirus in the family Paramyxoviridae, is a highly contagious pathogen, affecting various domestic, and wild carnivores. Conventional methods are too cumbersome to be used for high-throughput screening of anti-CDV drugs. In this study, a recombinant CDV was rescued using reverse genetics for facilitating screening of anti-CDV drug in vitro. The recombinant CDV could stably express the NanoLuc® luciferase (NLuc), a novel enzyme that was smaller and “brighter” than others. The intensity of NLuc-catalyzed luminescence reaction indirectly reflected the anti-CDV effect of a certain drug, due to a positive correlation between NLuc expression and virus propagation in vitro. Based on such a characteristic feature, the recombinant CDV was used for anti-CDV assays on four drugs (ribavirin, moroxydine hydrochloride, 1-adamantylamine hydrochloride, and tea polyphenol) via analysis of luciferase activity, instead of via conventional methods. The result showed that out of these four drugs, only the ribavirin exhibited a detectable anti-CDV effect. The NLuc-tagged CDV would be a rapid tool for high-throughput screening of anti-CDV drugs.

scholarly journals Tet-Inducible Production of Infectious Zika Virus from the Full-Length cDNA Clones of African- and Asian-Lineage Strains

Viruses ◽  
2018 ◽  
Vol 10 (12) ◽  
pp. 700 ◽  
Author(s):  
Lizhou Zhang ◽  
Wei Ji ◽  
Shuang Lyu ◽  
Luhua Qiao ◽  
Guangxiang Luo
Keyword(s):  
Zika Virus ◽  
Reverse Genetics ◽  
Genetic System ◽  
Full Length ◽  
Reverse Genetic System ◽  
Cdna Clones ◽  
Viral Pathogen ◽  
Stable Plasmid ◽  
Asian Lineage ◽  
Delta Virus

Zika virus (ZIKV) is a mosquito-borne flavivirus that has emerged as an important human viral pathogen, causing congenital malformation including microcephaly among infants born to mothers infected with the virus during pregnancy. Phylogenetic analysis suggested that ZIKV can be classified into African and Asian lineages. In this study, we have developed a stable plasmid-based reverse genetic system for robust production of both ZIKV prototype African-lineage MR766 and clinical Asian-lineage FSS13025 strains using a tetracycline (Tet)-controlled gene expression vector. Transcription of the full-length ZIKV RNA is under the control of the Tet-responsive Ptight promoter at the 5′ end and an antigenomic ribozyme of hepatitis delta virus at the 3′ end. The transcription of infectious ZIKV RNA genome was efficiently induced by doxycycline. This novel ZIKV reverse genetics system will be valuable for the study of molecular viral pathogenesis of ZIKV and the development of new vaccines against ZIKV infection.

scholarly journals A Ferret Model of Canine Distemper Virus Virulence and Immunosuppression

2003 ◽  
Vol 77 (23) ◽  
pp. 12579-12591 ◽  
Author(s):  
Veronika von Messling ◽  
Christoph Springfeld ◽  
Patricia Devaux ◽  
Roberto Cattaneo
Keyword(s):  
Canine Distemper Virus ◽  
Reverse Genetics ◽  
Virulent Strain ◽  
Consensus Sequence ◽  
Disease Onset ◽  
Small Animal ◽  
Canine Distemper ◽  
Animal System ◽  
Leukocyte Number ◽  
Matrix Fusion

ABSTRACT Canine distemper virus (CDV) infects many carnivores, including ferrets and dogs, and is the member of the Morbillivirus genus most easily amenable to experimentation in a homologous small-animal system. To gain insights into the determinants of CDV pathogenesis, we isolated a strain highly virulent for ferrets by repeated passaging in these animals. Sequence comparison of the genome of this strain with that of its highly attenuated precursor revealed 19 mutations distributed almost evenly in the six genes. We then recovered a virus from a cDNA copy of the virulent CDV strain's consensus sequence by using a modified reverse genetics system based on B cells. We infected ferrets with this virus and showed that it fully retained virulence as measured by the timing of rash appearance, disease onset, and death. Body temperature, leukocyte number, lymphocyte proliferation activity, and cell-associated viremia also had similar kinetics. We then addressed the question of the relative importance of the envelope and other viral constituents for virulence. Viruses in which the envelope genes (matrix, fusion, and hemagglutinin) of the virulent strain were combined with the other genes of the attenuated strain caused severe rash and fever even if the disease onset was delayed. Viruses in which the nucleocapsid, polymerase, and phosphoprotein genes (coding also for the V and C proteins) of the virulent strain were combined with the envelope genes of the attenuated strain caused milder signs of disease. Thus, virulence-inducing mutations have accumulated throughout the genome.

scholarly journals Recovery of Recombinant Canine Distemper Virus That Expresses CPV-2a VP2: Uncovering the Mutation Profile of Recombinant Undergoing 50 Serial Passages In Vitro

2022 ◽  
Vol 11 ◽  
Author(s):  
Fuxiao Liu ◽  
Jiahui Lin ◽  
Qianqian Wang ◽  
Youming Zhang ◽  
Hu Shan
Keyword(s):  
Nonsense Mutation ◽  
Canine Distemper Virus ◽  
Reverse Genetics ◽  
Test Strip ◽  
Immunofluorescence Assay ◽  
High Morbidity ◽  
Missense Mutations ◽  
Canine Distemper ◽  
Reading Frame

Canine distemper and canine parvoviral enteritis are infections caused by the canine distemper virus (CDV) and canine parvovirus type 2 (CPV-2), respectively. They are two common infectious diseases that cause high morbidity and mortality in affected dogs. Combination vaccines have been broadly used to protect dogs from infections of CDV, CPV-2, and other viruses. VP2 is the most abundant protein of the CPV-2 capsid. It elicits potent immunity in animals and, therefore, is widely used for designing subunit antigen-based vaccines. In this study, we rescued a recombinant CDV (QN vaccine strain) using reverse genetics. The recombinant CDV (rCDV-VP2) was demonstrated to express stably the VP2 in cells for at least 33 serial passages in vitro. Unfortunately, a nonsense mutation was initially identified in the VP2 open reading frame (ORF) at passage-34 (P34) and gradually became predominant in rCDV-VP2 quasispecies with passaging. Neither test strip detection nor indirect immunofluorescence assay demonstrated the expression of the VP2 at P50. The P50 rCDV-VP2 was subjected to next-generation sequencing, which totally identified 17 single-nucleotide variations (SNVs), consisting of 11 transitions and 6 transversions. Out of the 17 SNVs, 1 and 9 were identified as nonsense and missense mutations, respectively. Since the nonsense mutation arose in the VP2 ORF as early as P34, an earlier rCDV-VP2 progeny should be selected for the vaccination of animals in future experiments.

scholarly journals Establishment of a reverse genetic system from a bovine derived Influenza D virus isolate

2021 ◽  
Author(s):  
Melle Holwerda ◽  
Laura Laloli ◽  
Manon Wider ◽  
Lutz Schönecker ◽  
Jens Becker ◽  
...  
Keyword(s):  
Genetic System ◽  
Influenza Viruses ◽  
Reverse Genetic System ◽  
Sequence Length ◽  
Reverse Genetic ◽  
Host Tropism ◽  
Culture Models ◽  
Conventional Cell ◽  
Viral Determinants

AbstractThe ruminant-associated Influenza D virus (IDV) has a broad host tropism and was shown to have zoonotic potential. To identify and characterize molecular viral determinants influencing the host spectrum of IDV, a reverse genetic system is required. For this, we first performed 5′ and 3′ rapid amplification of cDNA ends (RACE) of all seven genomic segments, followed by assessment of the 5′ and 3′ NCR activity prior to constructing the viral genomic segments of a contemporary Swiss bovine IDV isolate (D/CN286) into the bidirectional pHW-2000 vector. The bidirectional plasmids were transfected in HRT-18G cells followed by viral rescue on the same cell type. Analysis of the segment specific 5′ and 3′ non-coding regions (NCR) highlighted that the terminal 3′ end of all segments harbours an Uracil instead of a Cytosine nucleotide, similar to other Influenza viruses. Subsequent analysis on the functionality of the 5′ and 3′ NCR in a minireplicon assay revealed that these sequences were functional and that the variable sequence length of the 5′ and 3′ NCR influences reporter gene expression. Thereafter, we evaluated the replication efficiency of the reverse genetic clone on conventional cell lines of human, swine and bovine origin by use of an in vitro model recapitulating the natural replicate site of IDV in bovine and swine. This revealed that the reverse genetic clone D/CN286 replicates efficiently in all cell culture models. Combined, these results demonstrate the successful establishment of a reverse genetic system from a contemporary bovine IDV isolate that can be used for future identification and characterization of viral determinants influencing the broad host tropism of IDV.

scholarly journals Génétique inverse pour les virus à ARN à double brin

2009 ◽  
Vol 62 (2-4) ◽  
pp. 150
Author(s):  
H. Attoui ◽  
F. Mohd Jaafar ◽  
S. Maan ◽  
P. P.C. Mertens
Keyword(s):  
Reverse Genetics ◽  
Genetic System ◽  
Virus Genome ◽  
Dsrna Virus ◽  
Reverse Genetic System ◽  
Viral Dsrna ◽  
Viral Genomes ◽  
Virus Core ◽  
The Individual

Since the recognition by Sabin of the specific nature of reovi­ruses in 1959 and the characterisation of their genome as double-stranded ribonucleic acid (dsRNA) by Gomatos in 1960, there have been many attempts to rescue viruses by transfecting cells with viral dsRNA. These attempts were largely unsuccessful. In 1990, it was proposed by Roner and Joklik that messenger (m) RNA transcribed from orthoreovirus cores was infectious when that mRNA was transfected into cells together with rabbit reticu­locyte lysates that were pre-incubated with denatured viral dsR­NAs. However, attempts to reproduce these ‘rescue’ experiments failed in the hands of other scientists. A breakthrough came in 2007 when Kobayashi and Dermody established a plasmid-based reverse genetics system for the orthoreoviruses, which represents the first reliable, synthetic based, reverse genetic system for a dsRNA virus. The system made it possible to study the role of specified amino acids in the outer capsid protein by generating ‘designer’ mutants. A second and potentially even more signifi­cant breakthrough came in the same year when Boyce and Roy showed that mRNA transcribed from the bluetongue virus core was infectious, allowing rescue of the virus in BSR cells, a clone of BHK-21 cells. This same group extended their work to the generation of synthetic transcripts from complementary (c) DNA copies of each of the 10 genome segments cloned into plasmids, driven by the T7 polymerase. We report refinement of a T7 base transcription approach, which uses simple linear polymerase chain reaction (PCR) amplicons of the individual virus genome segments from any reovirus, for transcription of full length, fully capped mRNA transcripts. These transcripts were used to transfect BSR cells and permitted rescue of the corresponding viruses. There was no requirement for a second transfection of either capped or uncapped messages. The efficiency of the system was significantly improved, using modulators of the cells’ innate immunity; particu­larly, 2-aminopurine considerably enhanced the rescue and short­ened the time for appearance of lysis plaques. This system was successfully used for 12 segmented coltiviruses, the 12 segmented seadornaviruses and 10 segmented orbiviruses. The authors sug­gest that this simplified rescue-strategy should be applicable to any dsRNA virus, particularly the members of the 15 recognized genera of the family Reoviridae. The system is currently being tested for mono-partite dsRNA viral genomes.

scholarly journals Establishment of a Reverse Genetic System from a Bovine Derived Influenza D Virus Isolate

Viruses ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 502
Author(s):  
Melle Holwerda ◽  
Laura Laloli ◽  
Manon Wider ◽  
Lutz Schönecker ◽  
Jens Becker ◽  
...  
Keyword(s):  
Genetic System ◽  
Influenza Viruses ◽  
Reverse Genetic System ◽  
Sequence Length ◽  
Reverse Genetic ◽  
Host Tropism ◽  
Culture Models ◽  
Conventional Cell ◽  
Viral Determinants

The ruminant-associated influenza D virus (IDV) has a broad host tropism and was shown to have zoonotic potential. To identify and characterize molecular viral determinants influencing the host spectrum of IDV, a reverse genetic system is required. For this, we first performed 5′ and 3′ rapid amplification of cDNA ends (RACE) of all seven genomic segments, followed by assessment of the 5′ and 3′ NCR activity prior to constructing the viral genomic segments of a contemporary Swiss bovine IDV isolate (D/CN286) into the bidirectional pHW2000 vector. The bidirectional plasmids were transfected in HRT-18G cells followed by viral rescue on the same cell type. Analysis of the segment specific 5′ and 3′ non-coding regions (NCR) highlighted that the terminal 3′ end of all segments harbours an uracil instead of a cytosine nucleotide, similar to other influenza viruses. Subsequent analysis on the functionality of the 5′ and 3′ NCR in a minireplicon assay revealed that these sequences were functional and that the variable sequence length of the 5′ and 3′ NCR influences reporter gene expression. Thereafter, we evaluated the replication efficiency of the reverse genetic clone on conventional cell lines of human, swine and bovine origin, as well as by using an in vitro model recapitulating the natural replication site of IDV in bovine and swine. This revealed that the reverse genetic clone D/CN286 replicates efficiently in all cell culture models. Combined, these results demonstrate the successful establishment of a reverse genetic system from a contemporary bovine IDV isolate that can be used for future identification and characterization of viral determinants influencing the broad host tropism of IDV.

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