Cytopathic Effect Inhibition Assay for Canine Interferon Activity.

Authors:
  • NINoriko Mizukoshi Iwata
    Noriko Mizukoshi Iwata
  • TSToshiki Saito
    Toshiki Saito
  • KHKazushige Hamada
    Kazushige Hamada
  • YSYoshihiro Sokawa
    Yoshihiro Sokawa
  • SUSusumu Ueda
    Susumu Ueda
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Abstract
Conditions for cytopathic effect (CPE) inhibition assay of canine interferon (IFN) activity in Madin-Darby canine kidney (MDCK) cells and in canine tumor cell line A72 was investigated using the New Jersey strain of vesicular stomatitis virus (VSV). The culture supernatant from canine splenocytes stimulated with ultraviolet-irradiated Newcastle disease viruses was used as reference IFN. MDCK cells were resistant for growth of VSV when the cells were confluent. Full CPE was observed only in a sparsely growing culture. Canine IFN activity could be assayed on less than 10(4) MDCK cells/well of a 96-well microplate, and more than 10(5) TCID50/ml of VSV was required. In A72 cells, VSV growth was not as dependent on cell density as in MDCK cells, requiring 10(3) TCID50/ml of VSV. MDCK-VSV system showed a higher IFN sensitivity than A72-VSV, whereas reproducibility was higher for the latter than the former. Based on these findings, A72-VSV system for canine IFN assay is recommended for practical use due to its easy handling characteristics.