ABSTRACT
The prevalence of cancer in animals has increased significantly over the years. Mammary tumours are the most common neoplasia in dogs, in which around 50% are presented in the malignant form. Hence, the development and characterization of in vitro models for the study of canine tumours are important for the improvement of cancer diagnosis and treatment. Thus, the aim of this study was to characterize cell lines derived from canine mammary gland neoplasias which could be further used for basic and applied oncology research. Samples of canine mammary carcinomas were taken for cell culture and 2 cell lines were established and characterized in terms of cell morphology, tumourigenicity and global gene expression. Both cell lines presented spindle-shape morphology and shown common malignant features as in vitro invasion potential and expression of epithelial and mesenchymal proteins. Also, we found gene expression patterns between the 2 cell cultures in comparison to the normal mammary gland tissue. Cells from M25 culture showed a higher invasion and in vivo tumourigenic potential, associated to the overexpression of genes involved in focal adhesion and extracellular matrix communication, such as FN1, ITGA8 and THBS2. The phenotypic characterization of these cells along with their global gene expression profile potentially determine new therapeutic targets for mammary tumours.
Subject(s)
Dog Diseases/metabolism , Focal Adhesions/metabolism , Mammary Neoplasms, Animal/metabolism , Transcriptome , Animals , Cell Line, Tumor , Dog Diseases/pathology , Dogs , Extracellular Matrix/metabolism , Female , Gene Expression Profiling/veterinary , Mammary Neoplasms, Animal/pathology , Neoplasm InvasivenessABSTRACT
Bovine herpesvirus type 5 (BoHV-5) is an important pathogen that causes meningoencephalitis in cattle. Few studies have used the mouse as a model for BoHV-5 infection. Despite the fact that BoHV-5 can infect mice with immune deficiencies, little is known about viral replication, immune response, and the course of infection in the central nervous system (CNS) of wild-type mice. Therefore, the aim of this study was to evaluate the response in the CNS of BALB/c mice acutely infected with BoHV-5 at different days post-inoculation (dpi). BoHV-5, when inoculated intracranially, was able to infect and replicate within the CNS of BALB/c mice. Until 15 dpi, the mice were able to survive without showing prominent neurological signs. The infection was accompanied by a Th1 immune response, with a significant expression of the cytokines IFN-γ and TNF-α and chemokine CCL-2. The expression of these cytokines and chemokines was most significant in the early course of infection (3 and 4 dpi), and it was followed by meningoencephalitis with perivascular cuffing and periventriculitis, composed mainly of macrophages and lymphocytes. After the expression of cytokines and chemokine, the mice were able to curb BoHV-5 acute infection in the brain, since there was a decrease in the number of BoHV-5 DNA copies after 3 dpi and viable viral particles were not detected after 6 dpi. Importantly, BoHV-5 was able to infect the trigeminal ganglia during acute infection, since a large number of BoHV-5 DNA copies were detected on 1 and 2 dpi.(AU) i
Subject(s)
Animals , Cattle Diseases , Herpesvirus 5, Bovine , Virus Replication , Trigeminal Ganglion , Disease Susceptibility/immunology , Encephalitis , Cerebrum , Mice/immunology , Mice, Inbred BALB CABSTRACT
Bovine herpesvirus type 5 (BoHV-5) is an important pathogen that causes meningoencephalitis in cattle. Few studies have used the mouse as a model for BoHV-5 infection. Despite the fact that BoHV-5 can infect mice with immune deficiencies, little is known about viral replication, immune response, and the course of infection in the central nervous system (CNS) of wild-type mice. Therefore, the aim of this study was to evaluate the response in the CNS of BALB/c mice acutely infected with BoHV-5 at different days post-inoculation (dpi). BoHV-5, when inoculated intracranially, was able to infect and replicate within the CNS of BALB/c mice. Until 15 dpi, the mice were able to survive without showing prominent neurological signs. The infection was accompanied by a Th1 immune response, with a significant expression of the cytokines IFN-γ and TNF-α and chemokine CCL-2. The expression of these cytokines and chemokines was most significant in the early course of infection (3 and 4 dpi), and it was followed by meningoencephalitis with perivascular cuffing and periventriculitis, composed mainly of macrophages and lymphocytes. After the expression of cytokines and chemokine, the mice were able to curb BoHV-5 acute infection in the brain, since there was a decrease in the number of BoHV-5 DNA copies after 3 dpi and viable viral particles were not detected after 6 dpi. Importantly, BoHV-5 was able to infect the trigeminal ganglia during acute infection, since a large number of BoHV-5 DNA copies were detected on 1 and 2 dpi.
Subject(s)
Central Nervous System/immunology , Disease Susceptibility/immunology , Herpesviridae Infections/immunology , Herpesvirus 5, Bovine/immunology , Animals , Central Nervous System/virology , Cytokines/blood , Cytokines/genetics , Cytokines/immunology , Disease Models, Animal , Gene Expression Regulation/immunology , Herpesvirus 5, Bovine/physiology , Mice , Mice, Inbred BALB C , Trigeminal Ganglion/immunology , Trigeminal Ganglion/virology , Virus ReplicationABSTRACT
Intranasal inoculation of equid herpesvirus type-1 (EHV-1) Brazilian strains A4/72 and A9/92 induced an acute and lethal infection in four different inbred mouse strains. Clinical and neurological signs appeared between the 2nd and 3rd day post inoculation (dpi) and included weight loss, ruffled fur, a hunched posture, crouching in corners, nasal and ocular discharges, dyspnoea, dehydration and increased salivation. These signs were followed by increased reactivity to external stimulation, seizures, recumbency and death. The virus was recovered consistently from the brain and viscera of all mice with neurological signs. Histopathological changes consisted of leptomeningitis, focal haemorrhage, ventriculitis, neuronal degeneration and necrosis, neuronophagia, non-suppurative inflammation, multifocal gliosis and perivascular infiltration of polymorphonuclear and mononuclear cells. Immunohistochemical examination demonstrated that EHV-1 strains A4/72 and A9/92 replicated in neurons of the olfactory bulb, the cortex and the hippocampus. In contrast, mice inoculated with the EHV-1 Brazilian strain A3/97 showed neither weight loss nor apparent clinical or neurological signs; however, the virus was recovered consistently from their lungs at 3 dpi. These three EHV-1 strains showed distinct degrees of virulence and tissue tropism in mice. EHV-1 strains A4/72 and A9/92 exhibited a high degree of central nervous system tropism with neuroinvasion and neurovirulence. EHV-1 strain A3/97 was not neurovirulent despite being detected in the brains of infected BALB/c nude mice. These findings indicate that several inbred mouse strains are susceptible to neuropathogenic EHV-1 strains and should be useful models for studying the pathogenesis and mechanisms contributing to EHV-induced myeloencephalopathy in horses.
Subject(s)
Herpesviridae Infections/veterinary , Herpesvirus 1, Equid/pathogenicity , Horse Diseases/virology , Animals , Brain/pathology , Brain/virology , Female , Herpesviridae Infections/pathology , Herpesviridae Infections/virology , Horse Diseases/pathology , Horses , Mice , Models, Animal , Neurons/pathology , Neurons/virology , VirulenceABSTRACT
The ventilation method used in the management of laboratory rats is important in maintaining their health. Rats kept under general diluting ventilation (GDV) are exposed to high levels of pollutants present in the environment (dust, airborne bacteria, etc.) or those pollutants produced by animal metabolism and excretion inside the boxes (e.g. ammonia and carbon dioxide). These pollutants may contribute to respiratory pathologies. An alternative experimental ventilation system for laboratory animal housing using intracage ventilation technology (individually ventilated cage system, IVC) was developed. In this system, ammonia levels decreased and rats exhibited better reproductive performance and a lower incidence of pneumonia than rats maintained under GDV. Using two different levels of air speed (0.03-0.26 m/s: IVC(1); 0.27-0.80 m/s: IVC(2)), the effects of IVC were compared with GDV (control) in Wistar rats in terms of respiratory mucus properties, on the nasal epithelium (as measured by quantitative morphometry) and on the lungs (as determined by the cellular composition obtained by bronchoalveolar lavage). Mucus of the respiratory system was evaluated using the following techniques: rheology (viscoelasticity) by microrheometer, in vitro mucociliary transportability (frog palate) and contact angle (an indicator of adhesivity). Also, membrane transepithelial potential difference was measured as a biomarker of airway integrity. After bedding was changed, ammonia concentrations inside the cages on day 3 were significantly higher for GDV than for IVC(1) and IVC(2). The potential-difference values for IVC(1), IVC(2) and GDV in the epiglottis and in the trachea also showed differences. Although some significant differences were observed across the three groups in counts of some cell types, the intragroup results were highly variable among individuals and inconsistent between sexes. No significant differences in the other parameters were found across groups. These results establish that rats maintained under GDV in relatively unregulated conditions are exposed to factors that can lead to deleterious effects on the ciliated epithelium of the airways, and that these effects can be prevented by the use of IVC.
Subject(s)
Animal Welfare , Housing, Animal , Respiratory Tract Diseases/prevention & control , Rodent Diseases/prevention & control , Ventilation/methods , Air Pressure , Ammonia , Animal Husbandry/instrumentation , Animals , Animals, Laboratory , Brazil , Bronchoalveolar Lavage Fluid/cytology , Epithelium/metabolism , Epithelium/pathology , Humans , Inflammation/prevention & control , Male , Nose/pathology , Rats , Rats, WistarABSTRACT
The role of the pulmonary alveolar macrophages (PAM) in the lung defense mechanism was evaluated in horses infected with equine hespesvirus-1 (EHV-1). Five adult horses were exposed to 10(6.6) TCID50 EHV-1 by intranasal instillation. Cytology of bronchoalveolar lavage (BAL) was performed using cytocentrifugation of samples and slides stained by Rosenfeld. Cell concentration was adjusted to 2´10(6) cells/ml, for the measurement of macrophage activity - spreading, phagocytosis of zymosan particles and release of hydrogen peroxide (H2O2). All animals were positive in virus isolation on the second, third and fifth days post-inoculation (DPI). Seroconversion was observed on the 14th DPI. Lymphocytosis was observed by BAL cytology on the 16th DPI. Measurement of macrophage activity demonstrated a marked increase in the spreading rate, on the 23rd and 30th DPI. Phagocytosis was decreased on the second DPI, and returned to levels similar to those observed before inoculation on the 23rd DPI. The amount of H2O2 released by PAM declined on day 2, but, by day 16, they returned to values similar to those observed before inoculation. The decline in PAM activity in the acute phase of disease is indirect evidence that these cells have an important role in lung defense mechanisms against this agent.
Subject(s)
Animals , Bronchoalveolar Lavage , Herpesvirus 1, Equid , HorsesABSTRACT
Devido à importância dos macrófagos alveolares (MA) nos mecanismos de defesa pulmonar, foram realizados estudos para avaliar a atividade desses fagócitos em cavalos hígidos. Foram realizados lavados broncoalveolares (LBA) em cinco cavalos clinicamente sadios. A citologia foi realizada pela citocentrifugaçäo das ammostras e posterior confecçäo de lâminas coradas pelo método de Rosenfeld. Todas as amostras do LBA foram centrifugadas e a concentraçäo celular foi ajustada para 2x10 elevada a sexta potência células/ml, para a mensuraçäo da atividade macrofágica (testes de espraiamento, fagocitose e liberaçäo de peróxido de hidrogênio). A contagem diferencial das células presentes no LBA demonstrou a predominância de macrófagos (59,0ñ6,9 por cento). Os resultados obtidos nos testes de mensuraçäo da atividade macrofágica foram: índice de espraiamento 25,1ñ19,7 por cento, índice de fagocitose 89,4ñ6,2 por cento e liberaçäo de peróxido de hidrogênio 1,6ñ0,3nmoles/2x10 elevado a quinta potência células (sem PMA - phorbol 12-myristate 13-acetate) e 1,8ñ0,4nmoles/2x10 elevado a quinta potência células (com PMA). Os resultados demonstraram um padräo de atividade para MA de cavalos hígidos, os quais apresentaram índices de ativaçäo mesmo sem elicitaçäo prévia, indicando que as técnicas utilizadas foram adequadas para tal propósito
