Development of serological diagnosing methods for canine pemphigus by ELISA using fusion protein of canine desmosomal proteins

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The extraxcellular portion of canine desmosomal proteins, desmoglein 1, desmoglein3 and desmocollin 1 were cloned and sequenced, and fusion proteins were made by baculovirus expression in order to conserve the three dimensional structure of these proteins. Fusion proteins were confirmed to react with human pemphigus foliaceus sera which target human desmoglein 1, with human pemphigus vulgaris sera which target human desmoglein 3 by ELISA. These proteins were then measured the titer by ELISA with the sera from dogs with 6 pemphigus vulgaris, 1 paraneoplastic pemphigus, 1 pemphigus vegetans, 44 pemphigus foliaceus. Normal dog sera from 54 dogs and sera from dogs with the other dermatological disorders such as demodicosis and pyoderma were used as controls. Diagnosis of these disorders was based the following findings including characteristic clinical features, histopathological findings such as subcorneal large pustules with abundant neutrophils/eosinophils and acantholytic keratinocytes, intercellular fluorescent deposits by direct and indirect immunofluorescence test, the existence of autoantibodies in sera by living keratinocyte staining. Four out of six sera with pemphigus vulgaris showed a significant high titer in ELISA using recombinant desmoglein 3 compared with control sera. However, sera from pemphigus foliaceus were not consistently reacted with recombinant canine desmoglein 1.As a result, the serological diagnosing method for pemphigus vulgaris by ELISA using recombinant proteins of canine desmoglein 3 seems to be successful, but for pemphigus foliaceus, the ELISA is not yet a practical tool for the diagnosis of canine pemphigus foliaceus at the present time.