OBJECTIVE: To determine whether dogs with spontaneously-occurring diabetes mellitus demonstrate serological reactivity to proinsulin. SAMPLE POPULATION: Serum samples were collected from 15 newly-diagnosed diabetic, 15 insulin-treated diabetic and 15 non-diabetic control dogs. PROCEDURES: Canine proinsulin was cloned into a prokaryotic expression vector to generate recombinant poly-histidine-tagged protein in Escherichia coli. A Western blotting assay was developed for detection of proinsulin autoantibodies in canine sera. RESULTS: Reactivity to canine proinsulin was detected in 3 of 15 control dogs, 8 of 15 newly-diagnosed diabetic dogs and 6 of 15 insulin-treated diabetic patients. Of these reactors, only 1 control dog, 1 newly-diagnosed diabetic dog and 3 insulin-treated diabetic dogs recognised porcine insulin by ELISA, suggesting that the remaining proinsulin reactors might have been recognising proinsulin-specific epitopes. CONCLUSIONS AND CLINICAL RELEVANCE: This study suggests that proinsulin autoantibodies are present in a proportion of diabetic dogs. Further work is required to refine the assay and clarify the significance of these autoantibodies.
OBJECTIVE: To determine whether dogs with spontaneously-occurring diabetes mellitus demonstrate serological reactivity to proinsulin. SAMPLE POPULATION: Serum samples were collected from 15 newly-diagnosed diabetic, 15 insulin-treated diabetic and 15 non-diabetic control dogs. PROCEDURES: Canine proinsulin was cloned into a prokaryotic expression vector to generate recombinant poly-histidine-tagged protein in Escherichia coli. A Western blotting assay was developed for detection of proinsulin autoantibodies in canine sera. RESULTS: Reactivity to canine proinsulin was detected in 3 of 15 control dogs, 8 of 15 newly-diagnosed diabeticdogs and 6 of 15 insulin-treated diabeticpatients. Of these reactors, only 1 control dog, 1 newly-diagnosed diabeticdog and 3 insulin-treated diabeticdogs recognised porcine insulin by ELISA, suggesting that the remaining proinsulin reactors might have been recognising proinsulin-specific epitopes. CONCLUSIONS AND CLINICAL RELEVANCE: This study suggests that proinsulin autoantibodies are present in a proportion of diabeticdogs. Further work is required to refine the assay and clarify the significance of these autoantibodies.
Authors: Kerstin M Ahlgren; Tove Fall; Nils Landegren; Lars Grimelius; Henrik von Euler; Katarina Sundberg; Kerstin Lindblad-Toh; Anna Lobell; Åke Hedhammar; Göran Andersson; Helene Hansson-Hamlin; Åke Lernmark; Olle Kämpe Journal: PLoS One Date: 2014-08-25 Impact factor: 3.240
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