Avian reovirus replication in mononuclear phagocytes in chicken footpad and spleen after footpad inoculation
2015
Chen, Yu San | Shen, Pin Chun | Su, Bor Sheu | Liu, Tsung Ching | Lin, Cheng Chung | Lee, Long Huw
Circulating monocytes and tissue macrophages were suggested to be susceptible to avian reovirus (ARV) infection. To determine if ARV infects and replicates in mononuclear phagocytes (KUL01-positive cells), we infected 3-day-old specific-pathogen-free chickens with ARV strain 2408 by inoculation of the left footpad. The left footpads and spleens were collected for analysis at 1.5 and 2.5 d after inoculation. Replication of ARV in the footpad and spleen was demonstrated by detection of the viral protein σNS using immunohistochemical testing and viral S1 RNA expression by real-time quantitative polymerase chain reaction (qPCR). Furthermore, immunofluorescent double-staining assay of cytocentrifuged cells and cryosections of the footpad and spleen for the viral protein σNS and the surface marker recognized by monoclonal antibody (MAb) KUL01 indicated that KUL01-positive cells costained with MAb H1E1, which recognizes ARV protein σNS. In addition, more ARV S1 RNA was measured by qPCR in the KUL01-positive cell samples prepared from the footpad or spleen 1.5 d after inoculation compared with non-KUL01-positive cell samples. The amounts of ARV S1 RNA in the spleen were significantly lower (P < 0.05) than the amounts in the footpad 1.5 d after inoculation. The results suggest that ARV infects mononuclear phagocytes and then replicates within these cells before migrating to the spleen, where it infects and replicates in KUL01-positive cells.
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