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PubMed1993

In situ hybridization of virulent canine distemper virus in brain tissue, using digoxigenin-labeled probes.

Only a few hybridization experiments have been performed for detection of canine distemper virus (CDV) nucleic acid sequences in tissue cultures and in various tissues. Those published studies used probes derived from tissue culture-adapted CDV, and hybridization signals were not obtained in the CNS tissue, although infective CDV and viral antigen were detectable in this tissue. We developed probes complementary to virulent CDV and were able to detect viral RNA not only in primary brain cell cultures, but also in brain tissues, by use of in situ hybridization. Sensitivity of the test at least equaled that of immunohistochemistry. We applied digoxigenin-labeled, strand-specific RNA probes complementary to the nucleoprotein-coding viral nucleic acid sequence. Our results indicate that to detect CDV nucleic acid sequences in brain tissues, it is essential to use probes derived from the virulent virus.

Animals↗
PubMedDate not supplied

[Ultrastructural analysis of pathological changes in ferrets with experimental canine distemper].

Morphologic study of the lungs, liver, spleen, and kidneys of ferrets infected with canine distemper virus has been carried out. Electron microscopy revealed virus reproduction in bronchial epithelial cells, types I and II alveolocytes, bile duct epitheliocytes, and hepatocytes. Mononuclear phagocyte system cells infected with the virus were found in all the examined organs. The most expressed pathological changes were observed in the lungs.

Animals↗
PubMed1996

[RT-PCR: a tool for the generation of clones for research on canine distemper].

In the present work an adapted method of the reverse transcriptase polymerase chain reaction (RT-PCR) is described. This procedure was an important tool for cloning a variety of gene sequences derived from oligodendrocytes and canine distemper virus (CDV). Among other applications, the obtained clones were used to produce probes for in situ hybridization and to sequence the inserted DNAs. Due to these molecularbiological techniques new insights were gained into the pathogenesis of CDV-induced demyelination and CDV-persistence in the central nervous system.

Animals↗
PubMed1997

Inhibition of tyrosine hydroxylase expression within the substantia nigra of mice infected with canine distemper virus.

Experimental infection of mouse brain with a neuroadapted strain of canine distemper virus (CDV) leads to early acute encephalitis, followed by late neurological diseases such as motor pathologies (paralysis and turning behavior) or obesity syndrome. We have previously shown that, during the early stage of infection, CDV replicates transiently in selective structures of the brain including the substantia nigra, a structure known to play a critical role in motor control. In this study we demonstrate that CDV replication in the substantia nigra induces an early decrease in transcript level of tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine synthesis. The CDV infection of neuroblastoma cell culture, constitutively expressing TH, results in downregulation of TH transcription in the absence of cell death. In the few surviving mice with motor deficiencies, a pronounced decrease in TH expression is associated with a loss of dopaminergic cell bodies in the absence of any viral transcripts and proteins, suggesting that the initial CDV infection was sufficient to trigger irreversible neurodegenerative processes.

Animals↗
PubMed1998

Serum distemper virus and parvovirus antibody titers among dogs brought to a veterinary hospital for revaccination.

OBJECTIVE: To determine serum canine distemper virus (CDV) and canine parvovirus (CPV) antibody titers in healthy dogs brought to a veterinary hospital for revaccination. DESIGN: Case series. ANIMALS: 122 dogs. PROCEDURE: Serum antibody titers were measured by means of hemagglutination inhibition (CPV titers) or serum neutralization (CDV titers) at the time dogs were brought to the hospital for revaccination. All dogs had been vaccinated between 271 and 1,665 days previously. Dogs were grouped by age, breed (purebred vs mixed breed), sex, and weight to determine whether these factors were associated with antibody titers. Serum CPV titers > or = 1:80 and serum CDV titers > or = 1:96 were considered protective. RESULTS: Breed, sex, and weight were not significantly associated with serum CPV and CDV titers. Age was significantly associated with CPV titer, with younger dogs having higher titers, but was not associated with CDV titer. Thirty-three of 122 (27%; 95% confidence interval, 19.0 to 34.9%) dogs had a less-than-protective CPV titer. Twenty-five of 117 (21%; 95% confidence interval, 13.6 to 28.4%) dogs had a less-than-protective CDV titer. CLINICAL IMPLICATIONS: Results suggest that, on the basis of serum antibody titers, the current practice of annual revaccination of dogs against CPV and CDV infection should be maintained. Measurement of antibody titers to determine whether revaccination is truly needed would seem justifiable in those dogs that have previously had an adverse reaction to vaccination.

Age Factors↗
PubMed1976

Fatal vaccine-induced canine distemper virus infection in black-footed ferrets.

Four black-footed ferrets that were live-trapped in South Dakota and transported to the Patuxent Wildlife Research Center died within 21 days after vaccination with modified live canine distemper virus. Immunofluorescence, European ferret inoculation, virus isolation attempts, and serum-neutralization tests indicated insufficient attenuation of the vaccine for this species.

Animals↗
PubMed1981

Measles and canine distemper antibody. Presence in sera from patients with multiple sclerosis and matched control subjects.

Antibody to measles virus and canine distemper virus (CDV) was demonstrated in sera from patients with multiple sclerosis (MS) and from carefully matched control subjects. Elevated measles and CDV antibody titers were found in patients with MS when compared with the matched control subjects. The correlation between the measles and CDV antibody titers was quite high, suggesting that the antibody levels between the two viruses are very closely related. Based on the results of our study and a review of the literature, our conclusion is that the CDV antibody levels in patients with MS and matched control subjects are associated with occurrence of measles virus antibodies.

Antibodies, Viral↗
PubMed1981

Examination of the immunological relationship between measles virus and canine distemper virus using monospecific measles antisera.

Indirect immunofluorescence titrations were performed with measles virus, the Rockborn strain of canine distemper virus (CDV), and a large plaque variant of the Onderstepoort strain of CDV (Ond-LP) using monospecific antisera prepared against either the haemagglutinin (anti-HA), the haemolysin (anti-HL), or the ribonucleoprotein (anti-RNP) of measles virus. Tests with anti-HA showed that the Rockborn strain of CDV was more closely related to measles virus than Ond-LP. The ribonucleoprotein antigens of the CDV strains were closely related to each other but were both related to and distinct from measles virus RNP. The use of measles anti-HL serum demonstrated that CDV possesses an antigenically related acetone-sensitive component equivalent to the haemolysin of measles virus. Absorption of human convalescent serum with excess quantities of acetone-fixed CDV antigens had no effect on measles-specific anti-HA, HL, or RNP activity in the serum. Absorption with measles antigens on the other hand, totally removed all measles and CDV-specific HA and RNP activity. CDV was not neutralised by any of the monospecific antisera when tested either as individual antisera or as mixtures. Our results demonstrate the occurrence of antigenic variation between different strains of CDV, they also reveal unique antigenic determinants in both measles virus and CDV.

Antigens, Viral↗
PubMed1993

Prevalence of canine distemper antibodies in the pagetic population.

Recent molecular evidence has implicated canine distemper virus (CDV) as a possible aetiologic agent in Paget's disease. However, previous serological studies have shown no differences in levels of anti-CDV antibodies between Paget's patients and controls. In this study, the technique of enzyme linked immunosorbent assay was used to measure anti-CDV antibodies in a group of Paget's patients from the North West of England. Some patients were undergoing treatment with 3-amino-hydroxypropylidene (APD), and the pre-treatment levels of antibody were compared with those following treatment. With several patients, it was also possible to compare the antibody levels with results from in situ hybridisation studies. No significant difference was found between the levels of anti-CDV antibodies in patients and controls. However, several patients and some of the controls did have markedly elevated levels of anti-CDV antibody. Antibody levels remained fairly constant following treatment with APD, except for two patients who showed marked changes. The patients positive for CDV by in situ hybridisation had significantly lower levels of anti-CDV antibodies when compared with those that were negative by in situ hybridisation. These results suggest that if CDV does cause Paget's disease, anti-CDV antibodies play little or no part in the disease pathogenesis. High levels of anti-CDV antibodies in both Paget's patients and controls suggest that a canine virus can infect humans. The fact that those patients that had CDV transcripts detectable in their bone cells had low levels of anti-CDV antibodies might suggest failure to clear the virus during an initial infection.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1998

Modelling the spread of phocine distemper virus among harbour seals.

Data presented in earlier publications on the 1988 epizootic among seals in North West Europe show a pattern that is somewhat inconsistent with the predictions of the standard mathematical model of epidemics. We argue that for animals living in herds or colonies, such as seals, the mutual contact behaviour is such that models for the transmission of infectious diseases should be applied with special care for the distinction between numbers and densities. This is demonstrated by using a mechanistic description of the contacts among seals, which leads to a slightly different formulation of the model. Results of the analysis of this formulation are more in line with the data. The model introduced here can be applied to epidemics among all kinds of animals living in herds and in fact to any species with constant local density, independent of the total population size (i.e., occupying a variable area). Application of the traditional formulation, using different parameters for herds of different sizes, will give equally good results for non-lethal diseases. However, especially for diseases with a low R0 and high death rates, such as the phocine distemper virus (PDV) disease, the two model formulations give quite different results. Further analysis of the model is performed to determine the most important factors influencing such an epidemic. The survival of infected animals turns out to have a disproportionately great influence on the intensity of the epidemic. Therefore in the case of the PDV epizootic we conclude that marine pollution may not only have contributed to the high death rates, but, if so, it has intensified the epizootic as well.

Animals↗
PubMed2002

Detection and genetic characterization of canine distemper virus (CDV) from free-ranging red foxes in Italy.

Fragments of the genes encoding the haemoagglutinin (H) and the nucleocapsid protein (N) of a canine distemper (CDV)-like virus affecting a red fox (Vulpes vulpes) were sequenced and analysed. The CDV-like virus detected in the fox was found to be not dissimilar, in both the H and N gene, from other CDVs spreading in Italy, as well as all over the world, and phylogenetic analysis on the H protein-encoding gene allowed to include all the Italian CDVs in the H European genotype.

Animals↗
PubMed1993

Canine distemper virus L gene: sequence and comparison with related viruses.

We cloned the genomic RNA of canine distemper virus (CDV) and determined the nucleotide sequence of the large (L) protein-coding gene. The L gene is 6573 nucleotides long and contains a single open reading frame coding for a polypeptide of 2161 amino acids (MW 246,354). The precise 5' end of the viral genome consists of a 38-nucleotide leader region. The CDV L protein shows over 77% amino acid similarity with its morbilliform relative measles virus (MV) with nearly 67% of their amino acids conserved. The sequence homology of 11 negative strand viruses L proteins is compared and relatedness was found in the following decreasing order: CDV, MV, Sendai virus, parainfluenza virus type 3, simian virus 5, parainfluenza virus type 2, mumps virus, Newcastle disease virus, respiratory syncytial virus, vesicular stomatitis virus, rabies virus. The consensus sequence of proposed functional domains involved in L gene catalytic activities was well conserved in the CDV L protein.

Amino Acid Sequence↗
PubMed1993

Canine distemper terminal and intergenic non-protein coding nucleotide sequences: completion of the entire CDV genome sequence.

Sequences critical for the transcription and replication functions of canine distemper virus (CDV) RNA polymerase were analyzed. The sequence was obtained from polymerase chain reaction (PCR) products using either c-DNA clones from a genomic library as template or in most instances genomic CDV RNA. Clones coding for the precise 3'- and 5'-ends of the CDV genome were sequenced and the results confirmed by additional PCR experiments. The virtual identity of terminal sequences and spacing at the two noncoding ends speak to the importance of these areas in replication and transcription. The sequence for each of the CDV gene boundaries was defined and all were compared to related viruses. This report completes the sequence determination of the CDV genome.

Base Sequence↗
PubMed1994

Serotype-specific and canine distemper virus cross-reactive H-2Kk-restricted cytotoxic T lymphocyte epitopes in the measles virus nucleoprotein.

Immunization of C3H mice with a vaccinia virus (VV) recombinant expressing the measles virus (MV) nucleoprotein (NP) induces an H-2Kk-restricted cytotoxic T lymphocyte (CTL) response. With reference to the predicted peptide epitope motifs binding to this MHC class I molecule, we have used synthetic peptides derived from the primary sequence of the MV NP to establish the identity of the Kk-restricted CTL epitopes. Two octameric peptides, LDRLVRLI (aa 52-59) and VESPGQLI (aa 81-88) sensitized P815-Kk cells to lysis by MV NP-induced CTLs. In contrast to LDRLVRLI, the sequence VESPGQLI is also present in the primary sequence of the NP from the closely related canine distemper virus (CDV). In vitro stimulation of spleen cells from VV NP-immunized mice with the peptides showed that peptide VESPGQLI induced CTLs which could also lyse CDV-infected cells, whereas peptide LDRLVRLI could only lyse cells presenting the MV protein. Different concentrations of peptides were used, and the lysis efficiencies for both epitopes were shown to be of the same order. The value of predicative motifs for determining MHC class I CTL epitopes is discussed.

Amino Acid Sequence↗
PubMed1997

Phosphorylation of canine distemper virus P protein by protein kinase C-zeta and casein kinase II.

Transcription by nonsegmented negative-strand RNA viruses is mediated by the viral RNA-dependent RNA polymerase and transcriptional cofactor P. The P protein is activated by phosphorylation, an event initiated by cellular kinases. The kinase used differs among this group of RNA viruses; vesicular stomatitis virus and respiratory syncytial virus utilize casein kinase II (CKII), whereas human parainfluenza virus type 3 utilizes PKC isoform zeta (PKC-zeta) for activation of its P protein. To identify the cellular kinase(s) involved in the phosphorylation of the canine distemper virus (CDV) P protein, we used recombinant CDV P in phosphorylation assays with native kinase activities present in CV1 cell extracts or purified CKII and PKC isoforms. Here, we demonstrate that the CDV P protein is phosphorylated by two cellular kinases, where PKC-zeta has the major and CKII the minor activities. In contrast, the P protein of another member of the morbillivirus genus, measles virus, is phosphorylated predominantly by CKII, whereas PKC-zeta has only minor activity. Selective inhibition of PKC-zeta activity within CV1 cells eliminated permissiveness to CDV replication, indicating an in vivo role for PKC-zeta in the virus replication cycle. The broad tissue expression of PKC-zeta parallels the pantropic nature of CDV infections, suggesting that PKC-zeta activity is a determinant of cellular permissiveness to CDV replication.

Animals↗
PubMed1990

Ultrastructural and biochemical findings in brain cell cultures infected with canine distemper virus.

To study the pathomechanism of demyelination in canine distemper (CD), dog brain cell cultures were infected with virulent A75/17-CD virus (CDV) and examined ultrastructurally. Special attention was paid to the oligodendrocytes, which were specifically immunolabelled. In addition, cerebroside sulfotransferase (CST), an enzyme specific for oligodendrocyte activity was assayed during the course of the infection. Infection and maturation as well as CDV-induced changes were found in astrocytes and brain macrophages. Infection of oligodendrocytes was rarely seen, although CST activity of the culture markedly decreased and vacuolar degeneration of these cells occurred, resulting in their complete disappearance. We concluded that the degeneration of oligodendrocytes and demyelination is not due to direct virus-oligodendrocyte interaction, but due to CDV-induced events in other glial cells.

Animals↗
PubMed1978

Encephalitis following vaccination against distemper and infectious hepatitis in the dog. An optical and ultrastructural study.

A 4-month-old, male, healthy dog developed CNS-symptoms 10 days after the second vaccination with live, attenuated distemper and canine hepatitis virus. The animal was euthanized after 2 weeks of illness. Light and electron microscopic examination of the CNS showed a partly necrotizing encephalomyelitis with numberous intranuclear and intracytoplasmic inclusion bodies, and the presence of probable pseudomyxovirus nucleocapsids and of crystalloid and tubuloreticular aggregates. Although there was conformity of inclusion bodies and probable viral structures as revealed by electron microscopy, the latter showed a much wider distribution. In addition, viral structures of a different type were found in polymorphonuclear leukocytes and vascular endothelial cells. Several morphological criteria led to the supposition that they are Picornaviruses, or possibly Arenaviruses. It is thought that their role in the disease process was at best an indirect one.

Animals↗
PubMed1989

Evidence of oligodendrocyte infection and degeneration in canine distemper encephalomyelitis.

Canine distemper encephalomyelitis is an important neurological disease of the dog and is also of comparative medical interest. With some viral strains, demyelinating encephalomyelitis is seen; whether or not oligodendrocyte infection occurs has remained controversial. By examining very early white matter lesions unequivocal oligodendrocyte infection has been identified. Accordingly the direct effect of virus on oligodendrocyte viability must be weighed in considering the pathogenesis of this canine CNS infection.

Animals↗