Aims/hypothesis: Autoantibodies to the ADP ribosyl cyclase/cyclic ADP-ribose hydrolase CD38 have been suggested to be markers of autoimmunity in Type 1 and Type 2 diabetes. The aim of this study was to develop a fluid phase assay for population screening.
Methods: Human recombinant CD38 was cloned and expressed by in vitro transcription and translation for fluid phase radio-binding assay, as a fusion protein in COS7 cells for fluid phase immunoprecipitation, and as a fusion protein for western blot assays. Antibody binding to each recombinant protein was measured in sera from patients with Type 1 diabetes, Type 2 diabetes and control subjects.
Results: Immunoprecipitation of radio-labelled in vitro transcribed and translated CD38 was low in all sera, including monoclonal anti-CD38 antibodies, with no difference between patients and control subjects. Monoclonal antibodies to CD38, but not patient or control sera immunoprecipitated recombinant CD38 fusion protein expressed in COS7 cells. Antibody binding to recombinant CD38 in solid-phase western blot assay was detected in sera from 2% of patients with Type 1 diabetes, 6% of patients with Type 2 diabetes, and 8% of control subjects.
Conclusions: This study failed to detect diabetes relevant binding of antibodies to recombinant CD38 using liquid-phase methods. Formal comparison of anti-CD38 antibody detection between laboratories is suggested.
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