RESUMO
To gain further insight into the pathogenesis of classical swine fever (CSF), the changes induced by hog cholera (HC) virus in pulmonary intravascular macrophages (PIMs) were examined. Twelve pigs were inoculated by the intramuscular route with a virulent strain of HC virus (Quillota strain) and killed in groups of three at 4, 7, 10 and 14 days post-inoculation. Immunohistochemical and ultrastructural examination revealed HC virus infection in endothelial cells, PIMs, and interstitial and alveolar macrophages. In addition to viral replication, a predominant feature was the secretory activation of PIMs, characterized by expanded rough endoplasmic reticulum and hyperplastic Golgi complexes. The results obtained suggest that macrophage activation and the subsequent release of pro-inflammatory mediators play an important role in the pathogenesis of CSF.
Assuntos
Vírus da Febre Suína Clássica/fisiologia , Peste Suína Clássica/patologia , Pulmão/patologia , Macrófagos Alveolares/patologia , Animais , Contagem de Células , Peste Suína Clássica/etiologia , Vírus da Febre Suína Clássica/ultraestrutura , Endotélio Vascular/patologia , Endotélio Vascular/virologia , Feminino , Técnicas Imunoenzimáticas/veterinária , Pulmão/irrigação sanguínea , Pulmão/virologia , Macrófagos Alveolares/virologia , Masculino , Microscopia Eletrônica/veterinária , Organelas/ultraestrutura , Suínos , Proteínas do Envelope Viral/análiseRESUMO
Classical swine fever (CSF) was induced in 20 pigs by inoculation with a virulent strain of CSF virus to determine sequential changes (2, 4, 7, 10 and 14 days post-inoculation) in the number and morphology of splenic macrophages (red pulp and lymphoid marginal zone) and thus to assess the role of these cells in the pathogenesis of the disease. The first splenic cells to be infected with CSF virus were macrophages in the marginal zone followed by other macrophage populations. The initial phase of CSF was associated with an increase in splenic macrophage numbers in the marginal zone and a decrease in the red pulp. Subsequently, the numbers in the red pulp increased. The study suggested that infection, mobilization and apoptosis of splenic macrophages play an important role in the spread of CSF virus in vivo. Moreover, the secretory changes that occurred in macrophages in the initial phase of the infection suggested that macrophages release chemical mediators capable of modulating pathogenesis.
Assuntos
Vírus da Febre Suína Clássica/fisiologia , Peste Suína Clássica/patologia , Macrófagos/patologia , Baço/patologia , Animais , Antígenos Virais/imunologia , Biomarcadores/análise , Contagem de Células/veterinária , Peste Suína Clássica/etiologia , Peste Suína Clássica/metabolismo , Vírus da Febre Suína Clássica/isolamento & purificação , Vírus da Febre Suína Clássica/ultraestrutura , Vesículas Citoplasmáticas/ultraestrutura , Feminino , Técnicas Imunoenzimáticas/veterinária , Corpos de Inclusão Viral/ultraestrutura , Macrófagos/metabolismo , Masculino , Fatores de TempoRESUMO
To study the effect of hog cholera virus on the epithelial cells of the bronchiolar mucosa, 12 pigs were inoculated with a highly virulent strain. Immunohistochemical and ultrastructural examination of the ciliated epithelial cells demonstrated an increase in the number of atypical cilia. The latter showed alterations in the microtubular pattern, possibly resulting from viral interference with the normal metabolism of the epithelial cells.
Assuntos
Brônquios/ultraestrutura , Cílios/ultraestrutura , Vírus da Febre Suína Clássica/fisiologia , Peste Suína Clássica/patologia , Animais , Brônquios/virologia , Vírus da Febre Suína Clássica/imunologia , Vírus da Febre Suína Clássica/ultraestrutura , Epitélio/ultraestrutura , Epitélio/virologia , Feminino , Técnicas Imunoenzimáticas , Masculino , Microscopia Eletrônica , Mucosa Respiratória/ultraestrutura , Mucosa Respiratória/virologia , Suínos , Proteínas do Envelope Viral/análise , Replicação ViralRESUMO
Twelve pigs were inoculated with a highly virulent strain of hog cholera virus (HCV) to study viral infection of megakaryocytes in the bone marrow and spleen. Immunohistochemical and ultrastructural examination revealed HCV infection in a small proportion (2.5-9.0%) of these cells from the 2nd to the 9th day after inoculation, at which time the experiment was terminated. Megakaryocyte infection accounts for the presence of viral antigens in platelets. The latter may represent a passive vehicle for spreading the virus in the animal.
Assuntos
Medula Óssea/patologia , Vírus da Febre Suína Clássica/fisiologia , Vírus da Febre Suína Clássica/ultraestrutura , Peste Suína Clássica/virologia , Megacariócitos/virologia , Baço/patologia , Animais , Antígenos Virais/análise , Medula Óssea/virologia , Contagem de Células , Peste Suína Clássica/patologia , Feminino , Técnicas Imunoenzimáticas/veterinária , Corpos de Inclusão/ultraestrutura , Masculino , Megacariócitos/ultraestrutura , Contagem de Plaquetas/veterinária , Baço/virologia , SuínosRESUMO
An infected mode between the Thiverval strain and Chinese strain of CSFV and MPK cells are established. The morphological structure and processing of different strains(T strain, C strain and F strain) of CSFV were studied by ultra-thin section electron microscopy. The virions of CSFV are roughly round and approximately 70 nm in diameter with a 40 nm core, and are wraped by membrane. The distinctive pathway of the maturation and release of CSFV are observed. Virons on different deverloping state and the pathologic changes of host cells have also been reported in this paper.
Assuntos
Vírus da Febre Suína Clássica/ultraestrutura , Animais , Linhagem Celular , Vírus da Febre Suína Clássica/fisiologia , Microscopia Eletrônica , Suínos , Porco MiniaturaRESUMO
Some characteristics of the multiplication of classical swine fever virus(CSFV) Thiverval strain were studied by means of the immunofluorescence technique. Under optimum culture conditions, the concentration of CSFV in the culture liquid multiplicated by MPK cells is ten times higher than by PK-15 cells. The half-life period of CSFV at 37 degrees C is 3 hour. The location of CSFV's replication in host cells is detected by the monoclonal antibodies of the structural protein E2 and the non-structural protein p120 of CSFV. Based on the above results, the ultrastructure of CSFV particles inside host cells was further studied using electron microscope and the changes of the ultrastructure of CSFV-infected cells are shown. The possible reasons for lower viral titre are also discussed in this paper.
Assuntos
Vírus da Febre Suína Clássica/fisiologia , Replicação Viral , Animais , Linhagem Celular , Vírus da Febre Suína Clássica/ultraestrutura , Imunofluorescência , Meia-Vida , Microscopia Eletrônica , Suínos , Temperatura , Vacinas Virais , Cultura de VírusRESUMO
Extracellular African swine fever (ASF) virus particles were specifically agglutinated by several lectins, suggesting the presence of surface glycosylated component(s) containing at least glucose, mannose, or both; galactose, N-acetylgalactosamine, or both; N-acetylneuraminic acid and N-acetylglucosamine, but not fucose. When virions were purified from infected Vero cells labeled with [14C]glucosamine, [14C]galactose and analyzed by polyacrylamide gel electrophoresis, no major structural glycoproteins were detected. However, several species of glycolipids were found when virions were extracted with organic solvents and analyzed by thin layer chromatography. These, plus two minor glycosylated structural components, of apparent mol wt 230K and 95K, could account for the agglutination of ASF virions with concanavalin A.
Assuntos
Vírus da Febre Suína Clássica/análise , Aglutinação , Carboidratos/análise , Vírus da Febre Suína Clássica/ultraestrutura , Glicolipídeos/análise , Glicoproteínas/análise , Lectinas , Receptores de Concanavalina A/análiseRESUMO
The early interactions between African swine fever virus (ASFV) and monkey kidney cells in culture, and the effect of chloroquine were studied by electron microscopy. Our results indicate that ASFV uptake occurs by endocytosis: after attachment to the cell surface, the virions were seen in coated pits and were internalized by endocytosis in endosomes and finally in lysosomes. Virions in coated vesicles were never seen. All these steps were completed in about 15 min. Direct penetration of viruses through the plasma membrane was never observed. In order to elucidate the participation of an acidic intracellular compartment in the penetration of ASFV, we studied the effect of chloroquine, a lysosomotropic drug known to increase the pH of acidic intracellular vacuoles and to inhibit ASFV infection. In the presence of this drug there were no apparent alterations on binding, endocytosis and intracellular distribution of the viral particles. The main effect of chloroquine was to retain the virions in lysosomes. When the drug was removed from the medium, the viral particles disappeared and images of binding of viral membranes with the membranes of the intracellular vacuoles were obtained, suggesting that the inhibited step is the uncoating of the virus. Viral fusion with the plasma membrane was obtained when the medium was acidified to pH 5-6. These results suggest that ASFV enters the cells by adsorptive endocytosis and that the uncoating process takes place intracellularly in a way similar to that described for Semliki Forest virus and other enveloped viruses.
Assuntos
Vírus da Febre Suína Clássica/fisiologia , Animais , Linhagem Celular , Membrana Celular/ultraestrutura , Chlorocebus aethiops , Cloroquina/farmacologia , Vírus da Febre Suína Clássica/efeitos dos fármacos , Vírus da Febre Suína Clássica/ultraestrutura , Rim , Microscopia Eletrônica , Receptores Virais/fisiologiaRESUMO
The glycoproteins E(rns) of classical swine fever virus (CSFV) and E(rns) and E2 of bovine viral diarrhoea virus (BVDV) are shown to be located at the surface of infected cells by the use of indirect immunofluorescence and by cytofluorometric analysis. The positive immunostaining of the cell surface was further analysed by immunogold electron microscopy and it could be shown that only extracellular virions were labelled. Gold granules were not seen at the cellular plasma membrane. In contrast to BVDV E2, the CSFV E2 of virions sticking to the plasma membrane was not accessible to the respective monoclonal antibodies. However, CSFV particles isolated from culture supernatant were able to bind both monoclonal anti-E(rns) and anti-E2 antibodies. For CSFV and BVDV, binding of anti-E(rns) antibodies to the virions was more pronounced than that of anti-E2. This finding was unexpected since E2 is considered to be the immunodominant glycoprotein.
Assuntos
Membrana Celular/virologia , Vírus da Febre Suína Clássica/fisiologia , Vírus da Diarreia Viral Bovina/fisiologia , Proteínas do Envelope Viral/análise , Vírion/fisiologia , Animais , Anticorpos Monoclonais , Bovinos , Células Cultivadas , Vírus da Febre Suína Clássica/isolamento & purificação , Vírus da Febre Suína Clássica/ultraestrutura , Vírus da Diarreia Viral Bovina/isolamento & purificação , Vírus da Diarreia Viral Bovina/ultraestrutura , Citometria de Fluxo , Técnica Indireta de Fluorescência para Anticorpo , Microscopia Imunoeletrônica , Proteínas do Envelope Viral/imunologia , Vírion/isolamento & purificação , Vírion/ultraestruturaRESUMO
Twenty pigs were inoculated with a virulent isolate (Quillota strain) of classical swine fever (CSF) virus to determine the chronological development of lesions in bone marrow. Histopathologic, ultrastructural and immunohistochemical (detection of viral antigen gp55, myeloid-histiocyte antigen, CD3 antigen, and FVIII-rag), and morphometric techniques were employed. Viral antigen was detected from 2 days postinfection (dpi) in stromal and haematopoitic cells, and severe atrophy related to apoptosis of haematopoitic cells was observed. Megakaryocytes (MKs) did not show significant changes in number, but there were important qualitative changes including 1) increased numbers of cloud-nuclei MKs, microMKs, apoptotic MKs, and atypical nucleated MKs and 2) decreased number of typical nucleated MKs. Morphometric study of these cells showed a decrease in cytoplasmic area. MK infection was detected from 2 dpi, but in a small percentage of cells. Myeloid cells showed quantitative changes, with an increase in granulocyte numbers. Apoptosis of lymphocytes and viral infection of erythroblasts were also observed. The main changes in stroma were depletion of T lymphocytes in the middle phase of the experiment and macrophages. Viral infection was also observed in these cells. MK lesions suggest dysmegakaryocytopoiesis, which would aggravate the thrombocytopenia already present and could be responsible for it. Granulocyte changes would lead to the appearance of circulating immature forms, whereas lymphocyte apoptosis in bone marrow would contribute to lymphopenia.