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PubMed1990

Sequence characterization of the membrane protein gene of paramyxovirus simian virus 5.

The complete nucleotide sequence of the membrane (M) protein gene of the paramyxovirus simian virus 5 (SV5) was determined from cDNA clones of viral mRNAs. The M gene boundaries were determined by (i) primer extension sequencing on M mRNA; (ii) nuclease S1 analysis; and (iii) primer extension sequencing on viral genomic RNA. The M gene mRNA consisted of 1371 templated nucleotides. It contains a single large open reading frame that can encode a protein of 377 amino acids with a predicted Mr = 42,253. The authenticity of the predicted M protein coding sequence was confirmed by synthesis of the M protein from mRNA synthesized from cDNA. The predicted M amino acid sequence indicated it is an overall hydrophobic protein carrying a net positive charge. Alignment of the SV5 protein amino acid sequence with the M protein sequences of other paramyxoviruses indicated that these viruses fall into the following two groups: (1) SV5, mumps virus, and Newcastle disease virus; or (2) Sendai, parainfluenza virus type 3, measles virus, and canine distemper virus, with mumps virus M sequence being the most closely related to SV5.

Amino Acid Sequence↗
PubMed1990

Sequence analysis of the phosphoprotein (P) genes of human parainfluenza type 4A and 4B viruses and RNA editing at transcript of the P genes: the number of G residues added is imprecise.

We cloned and sequenced the cDNAs against genomic RNAs and mRNAs for phosphoproteins (Ps) of human parainfluenza virus types 4A (PIV-4A) and 4B (PIV-4B). The PIV-4A and -4B P genes were 1535 nucleotides including poly(A) tract and were found to have two small open reading frames, neither of which was apparently large enough to encode the P protein. A cluster of G residues was found in genomic RNA and the number of G residues was 6 in both PIV-4A and -4B. However, the number of G residues at the corresponding site in the mRNAs to the genomic RNA was not constant. Three different mRNA cDNA clones were obtained; the first type of mRNA encodes a larger (P) protein of 399 amino acids, the second type encodes V protein of 229 or 230 amino acids, and the third type encodes the smallest protein (156 amino acids). Comparisons on the nucleotide and the amino acid sequences P and V proteins between these two subtypes revealed extensive homologies. However, these homology degrees are lower than that of NP protein. The C-terminal regions of the P and V proteins of PIV-4s could be aligned with all other Paramyxoviruses, PIV-2, mumps virus (MuV), simian virus 5 (SV 5), Newcastle disease virus (NDV), measles virus (MV), canine distemper virus (CDV), Sendai virus (SV), and PIV-3. On the other hand, the P-V common (N-terminal) regions showed no homology with MV, CDV, SV, and PIV-3. Seven phylogenetic trees of Paramyxoviruses were constructed from the entire and partial regions of P and V proteins.

Amino Acid Sequence↗
PubMed1989

X-linked severe combined immunodeficiency in the dog.

This study documents the occurrence of a form of X-linked severe combined immunodeficiency (SCID) in the dog with clinical, immunologic, and pathologic features similar to those of X-linked SCID with B cells in man. The disease in the dog is characterized by growth retardation and increased susceptibility to bacterial and viral infections in young pups. Affected pups have all died or were euthanatized by 5 months with signs of canine distemper, infectious hepatitis, or bacterial pneumonia. Laboratory findings include normal numbers of circulating B lymphocytes and normal concentrations of serum IgM, but low to absent concentrations of serum IgG and IgA, indicating a defect in the terminal differentiation of IgG and IgA B cells into immunoglobulin-secreting plasma cells. This is supported by the failure of peripheral lymphocytes to produce IgG or IgA plaque-forming cells in response to polyclonal activation. There are low-to-normal numbers of circulating T cells, but a severely depressed blastogenic response to T cell mitogens. Postmortem findings include thymic dysplasia and hypoplasia of lymphoid tissue. Family studies and breeding experiments are consistent with an X-linked recessive mode of inheritance.

Animals↗
PubMed1982

[Production of rabies vaccine in animal diploid cells].

Modalities for production of inactivated rabies vaccine derived from diploid hamster cell cultures are reported. The inactivated concentrated virus, purified by zonal centrifugation, is utilised for the preparation of vaccines destinated to carnivores, either in the form of monovalent vaccine or associated with distemper and canine contagious hepatitis vaccines. The inactivated concentrated virus is utilised for the preparation of bovine vaccine. The procedure is compatible with industrial production. The results concerning safety and potency tests of the experimental lots are presented.

Animals↗
PubMed1992

Investigations of environmental conditions during cluster indicate probable vectors of unknown exogenous agent(s) of multiple sclerosis.

During the tail-end of an active cluster several environmental investigations indicated that wildbirds were very probably the vectors of the unknown exogenous agent of MS. Canine distemper and genetic-autoimmune theories were very definitely eliminated because of the unusual pattern of the cluster. Studies of several avian pathogens unveiled Marek's (MDV) and/or IBD (Gumboro) as the most likely candidates for exogenous agent of MS.

Animals↗
PubMed1994

Intrathecal synthesis of major immunoglobulin classes in inflammatory diseases of the canine CNS.

The IgG index, IgM and IgA contents in cerebrospinal fluid and serum were examined retrospectively using an enzyme-linked immunosorbent assay (ELISA) in 69 dogs with inflammatory central nervous system (CNS) diseases of various aetiologies. Fifteen normal dogs were used as controls. After measuring IgG and albumin contents in serum and cerebrospinal fluid (CSF), the IgG index was calculated according to the formula of Link and Tibbling to demonstrate intrathecal immunoglobulin synthesis. A surprisingly high number of animals with encephalitis, including dogs with protracted diseases such as chronic distemper encephalitis and granulomatous meningoencephalomyelitis, showed in addition to an elevated IgG production evidence of intrathecal IgM and IgA production. The highest values of intrathecal as well as systemic IgA levels were found in dogs suffering from steroid responsive meningitis-arteritis. It was concluded that the control of the humoral immune response in the brain differs from that in other tissues. Because of striking similarities between dogs and humans in respect to humoral neuroimmunological reactions, the dog can be considered to be a useful animal model for the study of the intrathecal humoral immune response.

Animals↗
PubMed1982

Radioimmunoassay of measles virus hemagglutinin protein G.

Guinea pig and rabbit antisera from animals immunized with purified measles virus hemagglutinin (G) protein were used to establish a solid-phase four-layer radioimmunoassay for quantitative measurement of the G protein. The sensitive of the assay was 2 ng of purified G protein, and 200 micrograms of protein from uninfected Vero cells neither decreased the sensitivity nor reacted non-specifically in the assay. Radioimmunoassay standard dose-response curves were established and unknown values interpolated from these using the logit program of a desktop computer. Using this procedure, a measles virus growth curve in infected Vero cells was determined by measurement of G protein production. Under these same conditions, hemagglutination was not sensitive enough to detect early hemagglutinin production. Viral antigens in canine distemper virus, Newcastle disease virus, parainfluenza viruses 1-4, simian virus 5, and respiratory syncytial virus-infected cell lysates did not cross-react in the radioimmunoassay. A small degree of cross-reactivity was detected with mumps viral antigens, both with Vero cell-derived (wild-type strain) and egg-derived (Enders strain) purified virus preparations and with a cell lysate antigen prepared from wild-type mumps virus-infected Vero cells.

Animals↗
PubMed1989

The mumps virus nucleocapsid mRNA sequence and homology among the Paramyxoviridae proteins.

The nucleotide sequence of mumps virus nucleocapsid protein (NP) mRNA has been determined from two overlapping cDNA clones and confirmed by partial sequencing of the mRNA and the genome. The mRNA contains 1844 nucleotides excluding poly(A) and encodes a protein of 553 amino acids with a calculated molecular weight of 61,792. A comparison of the mumps virus nucleocapsid protein sequence with that of other paramyxoviruses revealed a moderate degree of homology (33.1%) with the Newcastle disease virus (NDV) only. The nucleocapsid proteins of all paramyxoviruses studied to date, excluding that of the genus pneumovirus, have a conserved sequence of six amino acids (Ser-Tyr-Ala-Met-Gly-Val) except that of NDV which has a mismatch of two amino acids (Ser-Phe-Ala-Met-Gly-Met) in that sequence. In addition, there is another conserved region of seven amino acids (Phe-Ala-Pro-Gly-X-Tyr-Pro) in the nucleocapsid proteins of mumps virus, Sendai virus and parainfluenza virus type 3. The nucleocapsid proteins of measles virus and canine distemper virus (CDV) also have this conserved region but with three conservative amino acid changes.

Amino Acid Sequence↗
PubMed1989

Preparation and characterization of monoclonal antibodies to nucleocapsid protein N and H glycoprotein of rinderpest virus.

Fifteen stable mouse spleen cell myeloma hybrids (hybridomas) producing monoclonal antibodies to rinderpest virus proteins were produced. The specificity of these monoclonal antibodies was established by radioimmunoprecipitation followed by polyacrylamide gel analysis and immunofluorescence. Nine antibodies were specific for the surface glycoprotein H. All the nine clones showed inhibition of haemagglutination by measles virus. The antibodies from two clones (A7D2 and B2F6) neutralise infectious virus. Six clones produce antibodies reacting with the nucleocapsid protein N. Three antigenic sites designated I-III, with sites I and II partially overlapping, were topographically mapped on the H molecule by competitive binding assay. Similarly, two antigenic sites I and II were delineated on the N protein. The monoclonal antibodies were used to study the antigenic relationships of H and N proteins of rinderpest virus, measles virus and canine distemper virus.

Animals↗
PubMed1994

Acute toxoplasmosis in a gray fox (Urocyon cinereoargenteus).

Toxoplasmosis was diagnosed in a gray fox (Urocyon cinereoargenteus) found moribund in Lake Loundes Park, Mississippi. There was almost total necrosis of mesenteric lymph nodes, focal hemorrhage and necrosis in the brain. Lesions in all of these organs were associated with numerous Toxoplasma gondii tachyzoites. Tissue cysts were seen in the brain. Organisms reacted with anti-T. gondii serum in an avidin-biotin immunohistochemical test. There was no evidence of concurrent distemper or rabies virus infection. Hammondia heydorni-like schizonts and gamonts were seen in sections of intestinal epithelium.

Animals↗
PubMed1990

Comparison of rinderpest and peste des petits ruminants viruses using anti-nucleoprotein monoclonal antibodies.

Monoclonal antibodies (MAbs) were obtained using a purified preparation of the RBOK strain of a rinderpest vaccine virus. The cytoplasmic immunofluorescent staining test showed that these clones had specificity for the nucleoprotein (N) of the virus. Six clones which immunoprecipitated the N protein corroborated these results. Thirteen anti-N MAbs were used to compare geographically widespread rinderpest viruses (RPV) and peste des petits ruminants viruses (PPRV) to two other morbilliviruses, measles (MV) and canine distemper (CDV). The N protein antigen profiles of the 23 isolates determined by immunofluorescent staining and enzyme linked immunosorbent assay (ELISA) on infected cells enabled us to classify the strains into groups. A differential identification of the morbilliviruses can be made using one MAb or associations of the MAbs. The potential to distinguish between RPV and PPRV and between virulent and avirulent strains of rinderpest is of primary interest.

Animals↗
PubMed1990

Morbillivirus group: genome organisation and proteins.

The Paramyxoviridae family is divided into three genera: Paramyxovirus, Pneumovirus and Morbillivirus. In the last group, there are four closely related viruses which are seriously pathogenic for man and animals, and usually cause acute diseases. At least two of them (measles and canine distemper viruses) can cause a persistent infection which leads to a chronic disease of the nervous system that, in the end, is fatal. For a long time, the biochemical analysis of morbilliviruses was hampered by the high susceptibility of some of their proteins to proteolysis. With cloning and sequencing technology, more data on the biology of those viruses are now available.

Animals↗
PubMed1990

Characterisation of the European seal morbillivirus.

The ELISA test originally developed for the detection of serum antibodies to rinderpest virus has been shown to detect cross-reacting antibodies in sera of diseased common and grey seals. Analysis of sera collected from various seal populations is in progress to establish the correlation between different morbillivirus neutralisation tests, ELISA tests and the disease status of the animals. RNA purified from post-mortem tissues removed from diseased seals has been analysed by hybridisation with cloned cDNAs made to various genes of canine distemper, peste des petits ruminants, rinderpest and measles viruses. This study has confirmed the presence of RNA sequences characteristic of a morbillivirus, but shown that the virus is not identical with any known morbillivirus. Work is in progress to determine the nucleotide sequences of clones carrying inserts homologous to morbillivirus genes isolated from cDNA made on a template of infected seal tissue RNA.

Animals↗
PubMed1995

The sequence of the N and L genes of rinderpest virus, and the 5' and 3' extra-genic sequences: the completion of the genome sequence of the virus.

We have sequenced the nucleocapsid (N) and polymerase (L) genes of the vaccine strain of rinderpest, and the 5' and 3' terminal domains of the genome. Together with previously published data, this completes the sequence of the entire genome of rinderpest virus. The L gene is identical in length to that of measles virus, encoding a 2183 amino acid protein with a calculated molecular weight of 248,100. The L protein sequence of three morbilliviruses is highly conserved, greater than 76% of residues being identical or conserved in all sequences. The N protein was, as for other sequenced genes, essentially identical to that of the virulent parent. The viral genome is 15,881 bases in length, similar to that of measles virus and slightly longer than that of canine distemper virus. The terminal sequences of the genome and those at the gene boundaries were compared to the analogous regions of other morbilliviruses and representatives of related groups of paramyxoviruses.

Amino Acid Sequence↗
PubMed1995

Morbilliviruses in aquatic mammals: report on round table discussion.

A workshop was organised to ascertain the current situation with regard to morbillivirus infections in aquatic animals. The great interest generated by the discovery of these new virus infections in 1988 has to some extent abated but much high quality research has continued in this field as the workshop showed. There is some serological evidence that the viruses have continued to circulate in most areas since the initial epizootics. As to their origin, it appears that the most likely source of the European seal morbillivirus (PDV-1) is the North Atlantic and Artic seal populations. As to the origin of the Mediterranean dolphin morbillivirus and the morbilliviruses isolated from porpoises, there is serological evidence that the viruses are widespread in many cetacean species in the Atlantic and 93% of long-finned pilot whales (Globicephala melas) which mass stranded between 1982 and 1993 were morbillivirus seropositive. The epizootic in freshwater seals in Lake Baikal was unrelated to events in the European marine mammal populations. The virus which infected these animals (PDV-2) is indistinguishable from canine distemper field strains. Serological and molecular biological studies provided evidence for the presence of the virus in the seals, at least as late as the Summer of 1992 when the animals were last sampled.

Animals↗
PubMed2003

Correction of a large animal model of type I Glanzmann's thrombasthenia by nonmyeloablative bone marrow transplantation.

OBJECTIVE: The purpose of this study was to determine if nonmyeloablative bone marrow transplantation would induce stable hematopoietic chimerism that would correct the bleeding diathesis associated with type I Glanzmann's thrombasthenia (GT). METHODS: Three young dogs (less than 12 weeks of age) with GT were transplanted with DLA-matched bone marrow from littermates. Recipients received a sublethal dose (200 cGy) of total-body irradiation (TBI) prior to infusion with bone marrow (1-4 x 10(8) cells/kg). Recipient dogs were immunosuppressed with cyclosporine (15 mg/kg) and mycophenolate mofetil (10 mg/kg). Chimerism was determined by quantitation of donor microsatellite repeat polymorphisms in peripheral blood DNA and by flow cytometry to detect the presence of glycoproteins IIb and IIIa on platelets. Platelet function was assessed by a clot retraction test. RESULTS: One dog died one week posttransplant due to hemorrhage. Another dog died four weeks posttransplant from an unrecognized congenital heart defect and complications due to canine distemper virus infection. At the time of death, microsatellite analysis indicated 35 to 50% chimerism. Flow cytometry showed 20% of circulating platelets positive for glycoproteins IIb and IIIa. The third dog is alive and doing well approximately two years posttransplant. Hematopoietic chimerism has been sustained at 35 to 60% with approximately 30% of the platelets positive for glycoproteins IIb and IIIa. Platelet function is normal based on clot retraction. The animal does not have clinical signs of bleeding. CONCLUSIONS: These observations suggest that GT and perhaps other severe inherited platelet disorders can be corrected using nonmyeloablative bone marrow transplantation to establish partial chimerism with normal platelets in the platelet compartment.

Animals↗
PubMed2005

Acute toxoplasmosis in three wild arctic foxes (Alopex lagopus) from Svalbard; one with co-infections of Salmonella Enteritidis PT1 and Yersinia pseudotuberculosis serotype 2b.

Acute disseminated toxoplasmosis was diagnosed in three wild arctic foxes (Alopex lagopus) that were found dead in the same locality on Svalbard (Norway). The animals included one adult female and two 4-months-old pups. The adult fox was severely jaundiced. Necropsy revealed multifocal, acute, necrotizing hepatitis, acute interstitial pneumonia, and scattered foci of brain gliosis, often associated with Toxoplasma tachyzoites. One pup also had Toxoplasma-associated meningitis. In addition, the latter animal was infected with Yersinia pseudotuberculosis serotype 2b and Salmonella Enteritidis phage type 1 (PT1), which may have contributed to the severity of the Toxoplasma infection in this animal. The diagnosis of toxoplasmosis was confirmed by positive immunohistochemistry and detection of anti-Toxoplasma gondii antibodies in serum of all foxes. The animals were negative for Neospora caninum, canine distemper virus, canine adenovirus, and rabies virus on immunolabelling of tissue sections and smears.

Agglutination Tests↗
PubMed2004

Prior DNA vaccination does not interfere with the live-attenuated measles vaccine.

The currently used live-attenuated measles vaccine is very effective although maternal antibody prevents its administration prior to 6 months of age. We are investigating the ability of a DNA vaccine encoding the measles viral hemagglutinin, fusion and nucleoprotein to protect newborn infants from measles. Here, we show that a measles DNA vaccine protects juvenile macaques from pathogenic measles virus challenge and that macaques primed and boosted with this DNA vaccine have anemnestic antibody and cell-mediated responses after vaccination with a live-attenuated canine distemper-measles vaccine. Therefore, this DNA vaccine administered to newborn infants may not hinder the subsequent use of live-attenuated measles vaccine.

Animals↗