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Comparative Study
. 2008 Sep 5;50(1):34.
doi: 10.1186/1751-0147-50-34.

Comparative studies on the pathogenicity and tissue distribution of three virulence variants of classical swine fever virus, two field isolates and one vaccine strain, with special regard to immunohistochemical investigations

Katinka Belák  1 , Frank Koenen, Hans Vanderhallen, Christian Mittelholzer, Francesco Feliziani, Gian Mario De Mia, Sándor Belák
Affiliations
Comparative Study

Comparative studies on the pathogenicity and tissue distribution of three virulence variants of classical swine fever virus, two field isolates and one vaccine strain, with special regard to immunohistochemical investigations

Katinka Belák et al. Acta Vet Scand. .

Abstract

Background: The aim of this study was to compare the tissue distribution and pathogenicity of three virulence variants of classical swine fever virus (CSFV) and to investigate the applicability of various conventional diagnostic procedures.

Methods: 64 pigs were divided into three groups and infected with the highly virulent isolate ISS/60, the moderately virulent isolate Wingene'93 and the live attenuated vaccine strain Riems, respectively. Clinical signs, gross and histopathological changes were compared in relation to time elapsed post infection. Virus spread in various organs was followed by virus isolation, by immunohistochemistry, applying monoclonal antibodies in a two-step method and by in situ hybridisation using a digoxigenin-labelled riboprobe.

Results: The tissue distribution data are discussed in details, analyzing the results of the various diagnostic approaches. The comparative studies revealed remarkable differences in the onset of clinical signs as well as in the development of the macro- and microscopical changes, and in the tissue distribution of CSFV in the three experimental groups.

Conclusion: The present study demonstrates that in the case of highly and moderately virulent virus variants the virulence does not affect the pattern of the viral spread, however, it influences the outcome, the duration and the intensity of the disease. Immunohistochemistry has the advantage to allow the rapid detection and localisation of the virus, especially in cases of early infection, when clinical signs are still absent. Compared to virus isolation, the advantage of this method is that no cell culture facilities are required. Thus, immunohistochemistry provides simple and sensitive tools for the prompt detection of newly emerging variants of CSFV, including the viruses of very mild virulence.

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Figures

Figure 1
Figure 1
Tonsil. Experiment I, PID 8. Positive immunohistochemical staining. Tonsil. Experiment I, PID 8. Superficial-epithelial cells, macrophages and lymphoid cells staining intensely for CSFV antigen in the cytoplasm with a monoclonal antibody specific for glycoprotein E2 (WH 303). Immunohistochemistry; EnVision™ +HP mouse system. Magnification 540×.
Figure 2
Figure 2
Tonsil. Experiment II, PID 7. Positive immunohistochemical staining. Tonsil. Experiment II, PID 7. Immunoreactivity to WH 303 monoclonal antibody as a cytoplasmic rim in the crypt-epithelial cells. Immunohistochemistry; EnVision™ +HP mouse system. Magnification 540×.
Figure 3
Figure 3
Lymph node. Experiment II, PID 6. Positive immunohistochemical staining. Lymph node. Experiment II, PID 6. Immunoreactivity to WH 303 monoclonal antibody in the cytoplasm of the reticulocytes and macrophages. Immunohistochemistry; EnVision™ +HP mouse system. Magnification 540×.
Figure 4
Figure 4
Spleen. Experiment II, PID 7. Positive immunohistochemical staining. Spleen. Experiment II, PID 7. Immunoreactivity to WH 303 monoclonal antibody in the cytoplasm of reticulocytes and macrophages. Immunohistochemistry; EnVision™ +HP mouse system. Magnification 540×.
Figure 5
Figure 5
Lungs. Experiment II, PID 14. Positive immunohistochemical staining. Lungs. Experiment II, PID 14. Immunoreactivity to WH 303 monoclonal antibody in the cytoplasm of the bronchiolar epithelial cells. Immunohistochemistry; EnVision™ +HP mouse system. Magnification 1080×.
Figure 6
Figure 6
Brain, blood vessel. Experiment II, PID 12. Degenerative changes. Brain, blood vessel. Experiment II, PID 12. Degenerative changes (pyknosis and karyorrhexis) of the endothelial cells. Haematoxylin-eosin staining. Magnification 540×.
Figure 7
Figure 7
Tonsil. Experiment III, PID 8. Positive immunohistochemical staining Tonsil. Experiment III, PID 8. Immunoreactivity to WH 303 monoclonal antibody as a cytoplasmic rim in the crypt-epithelial cells. EnVision™ +HP mouse system. Magnification 540×.
Figure 8
Figure 8
Tonsil. Experiment II, PID 4. In situ hybridisation. Tonsil. Experiment II, PID 4. Intense hybridisation signal for CSFV nucleic acid in the cytoplasm of tonsillar crypt epithelial cells. In situ hybridisation; DIG-labelled riboprobe. Magnification 675×.
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