Supachart PANNEUM, Wilairat CHUMSING, Theera RUKKWAMSUK

Cocultivation of caprine arthritis encephalitis virus-infected macrophages with primary goat synovial cells

Primary goat synovial cells were developed to be cocultivated with caprine arthritis encephalitis virus (CAEV)-infectedmonocytes. Two CAEV-seropositive goats and one CAEV-seronegative goat were confirmed for viral particles in the blood using thePCR method. Thereafter, heparinized blood samples from the three goats were collected and isolated for monocytes, which were furthercultivated to differentiate into macrophages. Cultivated macrophages of 7 10 days old were cocultivated with the primary goat synovialcells for 7 days. Microscopic examination was performed every 2 days for multinucleated syncytial formation, and immunofluorescenceassay was applied using monoclonal antibody against CAEV on day 7 to detect viral particles. Results showed that the primary goatsynovial cells cultivated with CAEV-infected macrophages from the two seropositive goats had multinucleated syncytial formationwith positive immunofluorescence, while the cocultured macrophages from the seronegative goats showed normal appearance withnegative immunofluorescence. The multinucleated syncytial cells were prepared for transmission electron microscope examinationand the results indicated that CAEV particles were clearly identified. In conclusion, our developed primary goat synovial cells and thecultivating system were proven to be an appropriate isolation assay for CAEV, which was the first report of cultivation and isolation ofCAEV in goats in Thailand.

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