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Biomedical subjects

M J Appel

Publications and source records attributed to M J Appel.

At least 91 records · Page 5Linked to original sources

Hyporesponsiveness to dog interferon induction in vitro.

Sera from dogs that were hyporesponsive to interferon (IFN) induction blocked virus induction of IFN in spleen cells in vitro but not IFN action in dog kidney cells. Similarly, macrophages or their culture fluids were found to block IFN induction in lymphoid cells in vitro. Lymphoid cells deprived of phagocytic cells secreted IFN continuously for 4 days, whereas lymphoid cells in the presence of phagocytic cells produced IFN for only 1 day. It was speculated that the 'hyporeactivity factor' in the sera from hyporesponsive dogs was macrophage-derived.

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Virus-induced interferon production in canine lymphoid cell cultures.

Virus-induced interferon (IFN) production in canine lymphoid cells was studied, using Newcastle disease virus as principal inducer. It was found that spleen cells at a concentration of 5 X 10(6) cells/ml with Newcastle disease virus at a multiplicity of infection of 1, incubated at 37 C in 5% CO2 for 24 hours, produced highest titers of IFN. Among the lymphoid cells from different tissues, IFN production was in the order of spleen = bone marrow greater than thymus greater than mesenteric lymph node greater than or equal to peripheral blood lymphocytes. Macrophages did not produce IFN, and virus-induced IFN production in spleen cells did not depend on the presence of phagocytic mononuclear cells. These optimal conditions and macrophage independence are different from those of mitogen-induced IFN production in canine lymphoid cells.

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Isolation and characterization of a canine rotavirus.

Canine rotavirus particles were visualized by direct electron microscopy in the feces from a clinically normal dog. The virus was subsequently propagated in cell cultures; it was characterized and compared with rotaviruses from other species. Replication of the virus in cell culture was found to be less dependent upon trypsin than that of human, bovine and porcine rotaviruses. Reproducible, sharp-edged plaques of various sizes were produced by the canine rotavirus in an established cell line of fetal rhesus monkey kidney, MA 104, under overlays of carboxymethyl cellulose or agarose. Intracytoplasmic inclusion bodies of different sizes and shapes were produced in infected MA 104 cells. By plaque reduction neutralization assay, a two-way antigenic relationship was found between the canine (CU-1) and simian (rhesus MMU 18006 and SA-11) rotaviruses. The canine rotavirus had a one-way antigenic relationship with feline (Taka), bovine (NCDV), and porcine (OSU) rotaviruses.

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Interferon in cerebrospinal fluid. A marker for viral persistence of canine distemper encephalomyelitis.

Interferon (IFN) was measured in serum and cerebrospinal fluid (CSF) of dogs after experimental (intranasal) infection with different strains of virulent canine distemper virus (CDV). Viral strains employed produced neurological changes in dogs that range from acute inflammatory to subacute, delayed demyelinating encephalomyelitis. With few exceptions, first appearance of serum-IFN correlated with the first elevated body temperature 4 days post-infection (p.i.). By 16 days p.i. IFN had disappeared from the serum of all infected dogs. In contrast, IFN was constantly detectable in CSF in dogs with CDV infection of the central nervous system (CNS). It was first detected 5 days p.i., was continuously detectable during the variable preclinical phase and into the period when signs of acute or delayed encephalomyelitis were evident. Dogs from which CDV would be retrieved from CNS tissue at necropsy always had CSF-IFN (up to 56 days p.i.). In contrast, dogs that recovered from infection, substantiated at necropsy by minimal, resolving CNS lesions and non-detectable virus, had IFN in CSF demonstrable for only a brief post-inoculation period. CSF-IFN appears to be a valid marker for CDV persistence in the canine CNS and may have broader applications.

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Lymphocyte-mediated immune cytotoxicity in dogs infected with virulent canine distemper virus.

Immune lymphocyte-mediated cytotoxicity (ILMC) was evaluated in dogs after intranasal exposure to one of the following three virulent strains of canine distemper virus: Cornell A75/17, Ohio R252, and Snyder Hill. Cytotoxicity was tested with peripheral blood lymphocytes as effector cells and primary dog testicle cells that were matched for histocompatibility as target cells. A strong correlation was found between ILMC and the course of the infection. Dogs that succumbed to encephalitis with any of the strains had little or no ILMC, whereas dogs that recovered had the highest activity. In the intermediate range were dogs with a delayed or reduced ILMC which developed persistent but subclinical central nervous system infections. A significant difference in onset, peak, and duration of ILMC was observed in dogs infected with different strains of canine distemper virus. ILMC responses began at 14 days postinfection (p.i.), reached a peak at 21 to 28 days p.i., and returned to preinoculation levels by 63 to 70 days p.i. in canine distemper virus A75/17- and R252-infected dogs. In contrast, ILMC in canine distemper virus Snyder Hill-infected dogs began at 10 days p.i., peaked by 14 to 17 days p.i., and approached preinoculation levels by 28 days p.i. Antiviral immunity as measured by ILMC appears to be a critical factor in determining the outcome in canine distemper virus-infected hosts. Furthermore, for certain viral biotypes, a delayed ILMC response correlated with persistent infection of the central nervous system.

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Canine viruses and multiple sclerosis.

Serum and cerebrospinal fluid from multiple sclerosis (MS) patients and control subjects were tested and compared for presence and titer of neutralizing antibody against the most common canine viruses. Canine viruses included canine distemper virus (CDV), canine adenovirus 1 (CAV-1), canine parainfluenza virus (CPIV), canine herpesvirus (CHV), canine coronavirus (CCV), and canine parvovirus (CPV). Neutralizing titers against measles virus (MV) and human adenovirus 8 (HA8) were also tested. Significantly elevated (p less than 0.05) antibody levels in sera from MS patients were found only against MV and CDV, but this depended upon the study population and the method of evaluation. The CDV-neutralizing component in serum could be absorbed on MV-infected cells. Results of this study failed to establish a link between canine viruses and MS.

Adenoviridae↗

Intranasal vaccine trial for canine infectious tracheobronchitis (kennel cough).

Two field trials were conducted during periods of endemic (summer) and epizootic (winter) canine infectious tracheobronchitis activity to evaluate the efficacy of three intranasal vaccines in a closed commercial beagle breeding kennel. A trivalent vaccine containing Bordetella bronchiseptica, canine parainfluenza, and canine adenovirus-2 was administered at 3 weeks of age. The vaccine was 71.2% and 81.8% effective in decreasing the incidence of coughing during the winter and summer trials, respectively. The number of deaths was lower in each of the vaccine groups than in the placebo groups. No adverse reactions were observed with any of the intranasal vaccines.

Adenoviridae↗

Canine parvovirus in a commercial kennel: epidemiologic and pathologic findings.

A study was conducted at a large canine commercial kennel that had suffered serious losses due to epidemic diarrheal disease following the introduction of canine parvovirus (CPV) into the kennel population. Mortality was highest in weaned (9- to 12-week-old) puppies coincident with the decline in maternal antibody titers. Virus was present in fecal samples from both fatal and non-fatal cases. Intestinal lesions of varying severity were present in dogs that died. One case of myocarditis occurred in a 5-week-old puppy. Reproductive performance was unaffected by the introduction of canine parvovirus into the kennel population.

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Canine parvovirus infection in South American canids.

Canine parvovirus (CPV) infections occurred in 5 of 35 South American canids at the Department of Conservation (DC), a breeding facility of the National Zoological Park in Front Royal, Va. The clinical signs were anorexia, lethargy, diarrhea, and vomiting. Three of the affected canids survived and had high hemagglutination-inhibition titers to CPV in the recovery period. Necropsy of the 2 that died revealed extensive necrosis of the intestinal mucosa; CPV particles were observed by electron microscopy in the intestinal contents of both animals. Six of the 30 canids that remained healthy had high hemagglutination-inhibition titers to CPV prior to the episode of illness, indicating earlier subclinical exposure. Pet dogs belonging to DC personnel that were screened as a possible source of the infection had no evidence of disease. All canids (including pet dogs) on the DC grounds were vaccinated repeatedly with a killed feline panleukopenia virus product after the episode, with little or no effect on existing titers.

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Humoral immunity to canine distemper after immunization of dogs with inactivated and live measles virus.

Pairs of dogs were immunized with 2 injections of a) measles virus material containing hemagglutinin but no hemolysin [Tween 80 and ether (TE) treated] b) material containing both hemagglutinin and hemolysin (freeze-dried and heated at 95 degrees C for 30 minutes), and c) live measles virus. Two unimmunized dogs were used as controls. All 8 dogs were challenged with virulent canine distemper virus (CDV). The two unimmunized dogs contracted a lethal infection, whereas immunized animals only showed mild signs of infection. Preimmunization with TE treated antigen gave the weakest protection. The animals displayed fever between day 4 and 10 after challenge. All titers of preexisting antibodies were boosted and antibodies against the measles hemolysin and CDV neutralizing antibodies increased from undetectable to moderate levels. Non-infectious intact virus (b) gave a more pronounced antibody response than TE antigen but antibodies against the hemolysin could not be detected. There was a good protection against disease (limited fever response). Replication of CDV caused a raise of antibody titers to levels normally seen only under conditions of hyperimmunization. Live measles virus stimulated the production of antibodies against both envelope components, but in spite of this virulent CDV could multiply and boost the titers of all antibodies.

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Natural and immune cytolysis of canine distemper virus-infected target cells.

Natural and immune cytolysis of canine distemper virus (CDV)-infected target cells in vitro is described. Lymphocytes expressing natural cytotoxicity were found in specific-pathogen-free beagle dogs and in beagle-coonhound crosses before vaccination with CDV and indefinitely after vaccination, when the ephemeral immune lymphocyte-mediated cytotoxicity (ILMC) had declined. In contrast to the natural lymphocyte-mediated cytotoxicity, the ILMC was genetically restricted, could not be blocked by CDV-specific antibody, and was effective against measles virus-infected as well as CDV-infectd target cells. Lymphocyte populations were depleted of Fc receptor and surface immunoglobulin-bearing cells by rosetting techniques and tested in comparison. An antibody-dependent cell-mediated cytotoxicity was demostrated against CDV-infected target cells that were preincubated with CDV antibody when Fc receptor-bearing lymphocytes were not removed. The ILMC was measurable for approximately 10 days beginning at 6 days post-vaccination. In contrast, CDV antibody measured by virus neutralization and humoral cytotoxicity was detectable by 6 days postvaccination and persisted at peak levels for at least 5 months.

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Enzyme-linked immunosorbent assay for evaluation of antibody to canine distemper virus.

An enzyme-linked immunosorbent assay (ELISA) for canine immunoglobulin (Ig) G antibodies to canine distemper virus (CDV) was developed and its test results were compared with those of the serum-neutralization (SN) test. The sera of 273 random-source adult dogs were examined. The two tests had a high degree of correlation. Very few discrepancies occurred and most of these were at the lower limits of each test. When the sera were tested at 1:100 dilution, there was a 98% agreement between the ELISA and SN test. Titrated SN and IgG ELISA tests also were performed on sera from 77 dogs whose lifetime medical histories were known. The results showed excellent agreement between the tests. Only five of the 77 sera showed any discrepancies and these were at detection-threshold levels. To follow antibody development in two dogs experimentally infected with CDV, it was necessary to use an ELISA which detected both canine IgM and IgG. The ELISA detected CDV-specific IgM a week before the SN test results became positive. The IgM titers rose for 3 weeks and descended to lower levels about the 4th and the 5th weeks. The SN titers closely followed the IgM titers. The ELISA detected IgG antibody about the 5th and the 6th weeks of infection. Results of both the SN test and the IgG ELISA remained elevated through the 70th day of testing.

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Studies on viral leukoencephalomyelitis and swayback in goats.

Six kid goats with CNS disease were studied by a variety of techniques. All had morphological evidence of Viral Leukoencephalomyelitis of Goats. (VLG) characterized by demyelinating encephalomyelitis and pneumonitis. Attempts were made to isolate a virus from 5 of these goats by direct culture of CNS and visceral tissues. Syncytial formation was found in some lung, spleen and bone marrow cultures but syncytia were found to be free of complete virus by electron microscopy. When newborn goats were inoculated with syncytia forming cells, a visna-like virus was found in bone marrow cultures of 1 goat 8 days post inoculation. Bone marrow cells from a twin goat 3 months after inoculation produced syncytia without complete virus. it is suggested that complete virus might be present in goats shortly after infection, while persisting in an incomplete form. Three of the 6 goats studied also had lesions of swayback with diffuse encephalomyelopathy and low liver copper levels. Control copper levels in kids with non-neurological disease were variable, occasionally being as low as swayback animals. The experience of others that swayback in the goat and low copper status are poorly correlated is confirmed. Discrepancies inherent in assaying formalin preserved tissues for copper are described.

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Isolation and immunisation studies of a canine parco-like virus from dogs with haemorrhagic enteritis.

A newly recognised canine parvo like virus was isolated from faeces of dogs with haemorrhagic enteritis. Cell cultures from several species were susceptible to it. Virus infected cells could be demonstrated by staining with fluorescent antibody reagents (prepared against canine virus or feline panleucopenia virus) or by haemagglutination with pig or rhesus monkey red blood cells. Inhibition of haemagglutination by specific antiserum prepared in specific-pathogen-free beagles provided a convenient method for viral identification. Experimental inoculation of specific-pathogen-free beagles resulted in elevated body temperatures and caused lymphopenia lasting one to three days. Feline panleucopenia virus vaccines protected dogs against challenge with virulent canine parvo-like virus.

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