Naturally occurring canine distemper virus encephalitis: distribution and expression of viral polypeptides in nervous tissues

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Abstract
Formalin-fixed brain tissues from 16 dogs with naturally occurring canine distemper virus (CDV) infection were investigated immunohistochemically by a panel of eight monoclonal antibodies (mAb) directed against four structural CDV proteins. Three mAb recognizing different epitopes of the polymerase (P-1, P-2, P-3) protein, two clones identifying different epitopes on each, the nucleocapsid (NP-1, NP-2) and fusion (F-2, F-3) protein, and one mAb directed against the hemagglutinin (H-2) protein were used. The immunoreactivity of the clones was tested on formalin-fixed, paraffin-embedded Vero cells, which were lytically infected with the neurotropic R252 (R252-CDV) or the Onderstepoort (CDV/Ond) strain of CDV. Clones directed against the H-2 and F-3 epitope recognized CDV/Ond but not R252-CDV. The remaining six clones showed positive immunoreaction with both CDV strains. In vivo expression and distribution of the individual proteins and their epitopes varied substantially between animals and within lesions from the same animal. The NP-2 epitope showed positive immunostaining in all 16 cases. The P-2 epitope was demonstrated in 13, the NP-1 epitope in 12, the P-3 epitope in 9, and the P-1 epitope in 3 brains, but staining was severely reduced compared with the NP-2 epitope and restricted to areas with strong NP-2 expression. Immunostaining was prominent in early and subacute and reduced in chronic demyelinating lesions. mAb directed against the H and F protein showed no immunoreaction in diseased brains.