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In vitro and in vivo properties of the virus causing natural canine distemper encephalitis.

A group of dogs with naturally occurring canine distemper developed prodromal systemic symptoms followed by neurological disorders. The post-infection courses of these diseases lasted approximately 2 months. A varying degree of demyelination and inclusion body formation was found mostly in the cerebella of virologically confirmed cases with little or no inflammatory response. The distribution of canine distemper virus antigen coincided with the histopathological lesions. The animals had moderate to high neutralizing titres to the virus in their sera and a low level of interferon-like activity in their cerebrospinal fluids. Isolation of viruses was most successful by the cocultivation method for brain specimens, but was possible by the direct method using lung homogenates. In infected Vero cells, the isolates derived from brain caused the formation of distinct plaques consisting of multinucleate giant cells, but the isolates from lung induced a cytopathic effect mainly consisting of cell rounding which eventually spread throughout the culture. The former infection produced less extracellular virus than the latter. The synthesis of the viral surface proteins H and F, and of M, was markedly reduced compared with that of the internal viral proteins such as NP, P and L. The SDS-PAGE migration pattern of the P protein varied from case to case, but was similar when isolates from different tissues of the same case were compared. In the affected tissues, the amount of viral polypeptides decreased markedly relative to that of the NP and there was also an absolute decrease compared to their abundance in Vero cells. This decrease was more obvious in the brain than in the lung. The relevance of these results is discussed.

Animals↗

Humoral immune response in dogs with old dog encephalitis and chromic distemper meningo-encephalitis.

The humoral immune response in sera and cerebrospinal fluids (CSFs) of dogs with various forms of canine distemper virus (CDV)-induced encephalitis was assessed by immunoprecipitation of radiolabelled nucleocapsid, phosphoprotein, membrane (M), haemagglutinin and fusion proteins. Sera from vaccinated dogs and hyperimmune sera contained antibodies to all the above antigens. In two cases of old dog encephalitis the sera and CSFs showed a restricted response to the M protein of CDV, whilst in three other cases of old dog encephalitis, two cases of chronic distemper (meningo-) encephalitis and experimentally induced encephalitis the humoral immune response appeared to be directed primarily to the nucleocapsid, phosphoprotein and the M protein but not the haemagglutinin or fusion proteins. Precipitation of the M protein by most of the sera was observed only when the antigen had been prepared by in vitro translation.

Animals↗

The biological characterization of field isolates of canine distemper virus from Japan.

Eight isolates of canine distemper virus (CDV) were obtained from seven dogs suffering from distemper by co-cultivation of their mononuclear cells with a marmoset B lymphoblastoid cell line, B95a. Six of the seven dogs had received one or more vaccinations. All of the isolates readily proliferated in B95a cells, but were not completely neutralized by anti-CDV canine plasma, which had high neutralizing activity against the Onderstepoort laboratory strain of CDV. Furthermore, different reactivities of monoclonal antibodies (MAbs) against CDV were observed between the field isolates and laboratory or vaccine strains of CDV in immunofluorescence studies. Immunoprecipitation analysis using MAbs detected the haemagglutinin protein of each new field isolate as 69K, 75K and 155K forms, and the fusion protein as 64K and 65K forms; the corresponding proteins of the Onderstepoort strain were detected as 75K and 61K proteins respectively. It is apparent from these results that the new field isolates of CDV have very different antigenic properties from the Onderstepoort vaccine strain.

Animals↗

Aspects of canine distemper virus and measles virus encephalomyelitis.

Canine distemper (CD) is a frequently fatal, systemic morbillivirus infection in the dog and other carnivores: encephalomyelitis is the common cause of death. Susceptibility to canine distemper virus (CDV) is now recognized in a wide range of non-domestic animals, most recently in captive lions, tigers and leopards. Furthermore, closely related viruses have produced CD-like diseases in marine mammals. CDV induces an inclusion-body encephalomyelitis in the dog and demyelination is often a conspicuous feature. Myelin injury is associated with the presence of virus but the mechanism of demyelination remains incompletely understood. Oligodendrocyte infection may be defective, as has been shown in vitro. CDV and measles virus (MV) produce similar systemic disorders in their respective hosts but differ markedly in the frequency of central nervous system (CNS) involvement, and in the pathogenesis of the more common neurological sequelae. Both CDV and MV have been considered as multiple sclerosis agents, and the association of CDV with other human disease has been suggested.

Animals↗

Vaccine-associated canine distemper infection in a litter of African hunting dogs (Lycaon pictus).

Four, 57 days old, African hunting dog puppies (Lycaon pictus) from one litter died within three weeks following vaccination with modified-live canine distemper virus (CDV) and killed canine adenovirus type 1, canine parvovirus and Leptospira icterohemorrhagiae and canicola. 18 days post vaccination, the animals developed neurologic disease characterized by episodes of grand mal seizures and circling. Macroscopic, histological and immunohistochemical studies revealed acute systemic CDV infection with acute encephalopathy. Virus isolation attempts using primary dog kidney cells, lung macrophages and Vero cells were negative. Therefore, the question whether the infection was the result of vaccination or natural infection remains open. The benefits and risks regarding the use of modified-live CDV vaccines and killed canine distemper vaccines in exotic carnivores are briefly discussed.

Animals↗

Canine distemper virus induces apoptosis through caspase-3 and -8 activation in vero cells.

We investigated the signal-transduction pathway of canine distemper virus-Onderstepoort (CDV-Ond) vaccine strain-mediated apoptosis in Vero cells. Canine distemper virus-Onderstepoort at a multiplicity of infection (MOI) of 0.1 induced DNA fragmentation 48 h after infection. Immunofluorescence staining revealed that 57% +/- 4% of the CDV-N-protein-positive cells were terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-positive, and all TUNEL-positive cells were CDV-N-protein-positive, indicating that CDV-Ond induced apoptosis only in the infected cells. We also found that CDV-Ond infection induced activation of caspase-3 and caspase-8. In the semi-quantitative reverse transcription-polymerase chain reaction assay for apoptosis-related genes, the expression of mRNA of the death receptor, Fas, was also increased in CDV-Ond-infected cells. In contrast, the expressions of Bcl-2 and Bax, regulators for intrinsic apoptotic signaling through the mitochondria, did not change. These results suggest that CDV-Ond induced apoptosis by activating caspase-3, possibly through caspase-8 signaling rather than through p53/Bax-mediated, mitochondrial signaling in the infected cells.

Animals↗

Cytopathic effect of canine distemper virus in tissue culture.

A characteristic cytopathic effect was obtained with canine distemper virus (Onderstepoort strain) propagated in chick embryo fibroblast monolayer tissue culture. In limiting dilutions the lesions were focal. The titer at the 20th tissue culture passage was approximately 10(5) TCD(50)/ml. Cytopathogenicity was specifically inhibited by distemper immune serum. Minute plaques were produced under agar overlay.

Animals↗

Distemper: not a new disease in lions and tigers.

In light of recent canine distemper virus (CDV) epidemics, we set out to determine the historical significance of CDV infection in captive lions and tigers in Switzerland. The retrospective case material consisted of 42 lion and tiger necropsy cases from 1972 to 1992. Necropsy reports for all lions and tigers were reviewed. All existing paraffin tissues were immunohistochemically examined with a polyclonal antibody raised against CDV. The results for 19 of the 42 lions and tigers were classified as positive by immunohistochemistry; 23 results were negative or questionable. The results for four animals (three positive and one negative ) were further tested by in situ hybridization, and the results concurred with the immunohistochemistry findings. CDV infection of large cats is older and more widespread than previously thought. All large cats in captivity should be immunized even if canine distemper is not believed to be a problem for large cats in the area.

Animals↗

Phylogenetic characterization of canine distemper viruses detected in naturally infected dogs in North America.

In 2004, six puppies and one adult dog from a total of four premises were subjected to necropsy evaluation. For five of the seven dogs, disease caused by canine distemper virus (CDV) infection was suspected based on clinical signs. In all of the dogs, a diagnosis of CDV infection was established by the presence of compatible gross and histologic lesions, immunohistochemical labeling for CDV antigen, and detection of CDV RNA by reverse transcription-PCR. To further characterize the CDV strains detected in the four cases, complete gene sequences were determined for the hemagglutinin (H) and fusion (F) protein genes, while partial gene sequencing was performed for the phosphoprotein gene. A total of 4,508 bases were sequenced for the CDV strains detected from each of the four cases. Two cases were found to have identical sequences except for 2 bases in the intergenic region of the F and H genes. Phylogenetic analysis strongly suggested an evolutionary relationship between sequences detected in these two cases and those of phocine distemper virus 2 and two other strains of CDV not previously detected in the continental United States. Clear phylogenetic relationships were not established for viruses detected in the two additional cases; however, one strain showed similarity to CDV strains detected in a panda from China. Importantly, the three CDV strains detected were demonstrated to be genetically distinct from known vaccine strains and strains previously reported in the continental United States.

Amino Acid Sequence↗

Protection against canine distemper virus in dogs after immunization with isolated fusion protein.

Canine distemper virus attachment (hemagglutinin [H] equivalent) and fusion (F) antigens were purified by affinity chromatography with monoclonal antibodies. The purified antigens were used to immunize groups of three dogs. Radioimmune precipitation assays with sera from these animals showed that the F antigen preparation was pure and induced only an F polypeptide-specific antibody response but that the H antigen preparation had a slight contamination by the F antigen. Immunized animals were challenged with virulent canine distemper virus. Two animals in each group developed pronounced humoral and cellular immune responses after challenge. Among these infected animals, only the dogs immunized with H antigen developed symptoms, albeit mild. In contrast, three nonimmunized control animals developed severe disease, with a fatal outcome in two cases. The complete resistance against challenge in two dogs was interpreted to reflect in one case anti-F immunity and in the other case most likely a high level of anti-H immunity. It is suggested that the F antigen may be of particular interest for the development of morbillivirus and possibly other paramyxovirus subunit or synthetic vaccines, because it can induce immunity capable of blocking virus infection and in situations of virus replication prevent the emergence of symptoms.

Animals↗

Canine distemper infection associated with acute nervous signs in dogs.

Eight vaccinated dogs suddenly developed progressive ataxia, paresis or paralysis of short duration. A histopathological examination revealed a non-suppurative meningoencephalitis suggestive of a viral infection, and immunohistochemical examination confirmed the presence of canine distemper virus antigen in five of the dogs. Distemper had not been suspected from the clinical examination of the dogs.

Acute Disease↗

Distemper encephalitis in pups after vaccination of the dam.

A five-year-old labrador bitch which had whelped 10 pups three days previously was given booster vaccination against distemper, adenovirus, parvovirus, parainfluenzavirus and leptospirosis. Eighteen days later, signs of central nervous system disease developed in some of the pups, five of which were ultimately euthanased. The cause of the nervous disease was found to be canine distemper, and serological studies showed that the infection was limited to some members of the litter, suggesting that the vaccinal rather than a field virus was more likely to have been responsible.

Animals↗

Outbreak off canine distemper in vaccinated dogs in Finland.

Canine distemper reappeared in dogs in Finland in 1990 after a 16-year absence. In 1994 to 1995 an outbreak occurred in areas with a high density dog population which involved dogs vaccinated against distemper. The estimated total number of cases was at least 5000, and 865 cases were confirmed by indirect fluorescent antibody testing of 3649 epithelial cell samples. The signs recorded by veterinary clinicians ranged from conjunctivitis, pyrexia and anorexia to signs of respiratory and gastrointestinal illness, with an estimated mortality of 30 per cent. Of the confirmed cases 631 (73 per cent) were between three and 24 months of age; 487 of these had been vaccinated at least once and 351 (41 per cent) had a complete vaccination history. Of these 351 fully vaccinated animals the proportion of dogs vaccinated with the most popular vaccine was significantly higher than would have been expected by its market share. In total, 4676 serum samples were collected from healthy vaccinated dogs during the peak and decline of the outbreak and tested for the presence of virus neutralising antibodies. The decrease in the proportion of young dogs with antibody titres < 1/8 coincided with the decline and end of the outbreak during the spring and summer of 1995. It was concluded that a critical decrease in the population's immunity during 1990 to 1994 was a major reason for the outbreak in the summer of 1994 and that the ultimate test for vaccines is an outbreak of disease.

Animals↗

Canine distemper antibodies in lions of the Masai Mara.

Canine distemper virus (CDV) has been implicated in some recent deaths of lions, which showed clinical signs of distemper, in the the Serengeti plain. Similar clinical findings have since been reported in lions of the Masai Mara. Fifty-five per cent of serum samples obtained from wild lions of the Masai Mara have been found to contain neutralising antibody to CDV, indicating that they had been exposed to the virus. Adult orphan lions kept in captivity, were vaccinated with the live attenuated Onderstepoort strain of CDV. The results indicated that the vaccine is both safe and immunogenic, and may be potentially useful for the prophylactic vaccination of lions at high risk.

Animals↗

Infection of canine mononuclear leucocytes by measles virus: possible mechanism of protection from canine distemper.

Measles virus was shown to be infectious to canine lymphocytes from peripheral blood as well as from different lymphoid tissues, and the same held true for canine macrophage cultures prepared from peripheral blood. The susceptibility of these leucocytes to measles virus was comparable with that of canine distemper virus. These observations supported the suggestion that interference with canine distemper virus by measles could be a possible mechanism of the heterotypic immunity observed in dogs.

Animals↗

Mechanism of persistence with canine distemper virus: difference between a laboratory strain and an isolate from a dog with chronic neurological disease.

The mechanism of persistence in culture was different with canine distemper virus (CDV) isolated from a dog with chronic neurological disease (ODE; old dog encephalitis) compared to a laboratory strain (CDV/Ond). CDV/Ond persistence was achieved after seven undiluted passages while CDV/ODE-8, the recent isolate, showed immediate persistence in Vero cells. Both persistent infections resisted challenge with lytic CDV/Ond but not with unrelated vesicular stomatitis virus. The medium from CDV/Ond infection showed interference, whereas the medium from CDV/ODE-8 infection did not. Ultracentrifugation of the CDV/Ond supernatant effectively removed the defective interfering (DI) particles. Fluorescent microscopy showed the presence of CDV antigen in the cytoplasm of both types of persistently infected cells. By electron microscopy, 1% of the CDV/Ond cells and 23% of the CDV/ODE-8 cells showed distemper viral nucleocapsids. Persistence of CDV/Ond appears to be due to classical DI particles, whereas persistence of CDV/ODE-8 appears to be due to cell-associated particles. The persistence of CDV in dogs with ODE may be due to this cell-associated phenomenon.

Animals↗

Histologic and immunocytochemical characterization of canine distemper-associated metaphyseal bone lesions in young dogs following experimental infection.

The proximal metaphyses of the humerus of weanling gnotobiotic dogs experimentally infected with canine distemper virus (CDV) were investigated histologically and immunocytochemically between 4 and 41 days after infection. Viral antigen was demonstrated in hematopoietic marrow and bone cells at postinfection day (PID) 5 and PID 7, respectively. Between PID 8 and 27, CDV antigen was abundantly present in marrow cells, osteoclasts, and osteoblasts and less frequently in osteocytes. Immunopositive cells in both osseous tissues and bone marrow declined between PID 29 and PID 36 and were absent by PID 41. Chondrocytes of the growth plate were negative for viral antigen throughout the observation period. In bone, viral antigen was more frequently observed in bone cells of the primary spongiosa than in the secondary spongiosa. There was a strong correlation between occurrence of CDV antigen and osseous changes. Associated metaphyseal bone lesions were mild and most prominent between PID 8 and PID 32. Lesions consisted of necrosis of osteoclasts, which was associated with subsequent persistence of the primary spongiosa (growth retardation lattice). Atrophy and necrosis of osteoblasts and marrow cells were also noted. Infection of metaphyseal bone cells appears to be common in young dogs with experimental systemic distemper. Bone cell infection is preceded by infection of marrow cells, and infected bone cells may experience degeneration and necrosis. This subtle viral effect may result in defects in bone modeling in CDV-infected dogs.

Animals↗

Serological survey of canine distemper virus infection using enzyme-linked immunosorbent assay.

For detection of antibodies against canine distemper virus (CDV), an enzyme-linked immunosorbent assay (ELISA) was developed using a crude extract from cells infected with the Onderstepoort strain of CDV as antigen. Twenty-six sera from dogs experimentally vaccinated with the Snyder-Hill strain of CDV were compared by ELISA and a standard virus neutralization (NV) test. Since a good correlation between the titers obtained by both tests was observed, ELISA was considered to be a rapid and reliable method for a serological survey of CDV infection. When a total of 167 sera from dogs suspected of canine distemper under natural conditions were examined by the ELISA, 29 of the sera (17%) were found to have low VN antibody titers and high ELISA titers. The reason for the discrepancy in the titers was discussed.

Animals↗