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Insulin as a target antigen in autoimmune diabetes: a natural repertoire as the source of antibody response

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Abstract

A solid-phase immunoenzymatic technique with B1- or B29-biotinylated insulin coupled to avidincoated wells was used to characterize serum anti-insulin antibodies and to locate insulin antibody-producing B lymphocytes in different organs of mice. Low natural serum anti-insulin IgM and IgG antibodies were found in ten different healthy inbred strains of mice. Prediabetic nonobese diabetic (NOD) mice had significantly higher measurements than BALB/c mice (P<0.05). Anti-insulin IgM antibody-producing B lymphocytes were found in bone marrow and spleen of NOD mice and healthy strains of mice, but not in peripheral lymph nodes, thymus, blood or pancreas. B29-fixed insulin was more frequently recognized than B1-fixed insulin. There was no relationship to the MHC or to other immune markers. IgG insulin antibody-producing cells were not detected. IgG insulin antibody-producing cells appeared in the draining lymph node and in the blood 10 days after immunization with insulin. IgM insulin-recognizing cells in the spleen were reduced in number during the same period (P<0.05–0.01 for BALB/c, DBA2, B10.D2 and NOD), suggesting migration of these cells. This was tested by in vivo staining of spleens with the red-fluorescent membrane linker PKH-26 on day 7 after immunization. Cells from immunized lymph nodes were FACS-sorted on day 10. Insulin antibody-producing B lymphocytes with red-fluorescence were found, indicating a splenic origin. Examination of IgG subclasses showed preferential production of complement-fixing IgG2b in sera and lymph node cells of immunized NOD mice (P<0.05 vs BALB/c). We conclude that a natural repertoire of insulin recognition exists in bone marrow and spleen of mice. Hydrophobic epitopes around B1 (B29-fixed insulin) are more frequently recognized than hydrophilic epitopes around B29 (B1-fixed insulin), indicating a genetically fixed pattern of autorecognition. Insulin-recognizing cells from the spleen function as the source of insulin antibody response. Preferential occurrence of complement-fixing IgG2b in NOD mice could contribute to autoimmune-mediated β-cell damage.

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Authors and Affiliations

  1. Department of Endocrinology, University Hospital of Internal Medicine, Hugstetter Strasse 55, D-79106, Freiburg, Germany

    K. -G. Petersen & A. -N. Khalaf

  2. Deutsches Wollforschungsinstitut, Aachen, Germany

    V. Naithani, M. Fabry & H. Gattner

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  1. K. -G. Petersen
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  2. A. -N. Khalaf
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  3. V. Naithani
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  4. M. Fabry
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  5. H. Gattner
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Petersen, K.G., Khalaf, A.N., Naithani, V. et al. Insulin as a target antigen in autoimmune diabetes: a natural repertoire as the source of antibody response. Acta Diabetol 31, 66–72 (1994). https://doi.org/10.1007/BF00570537

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  • DOI: https://doi.org/10.1007/BF00570537

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