Pancreatic islet cell surface glycoproteins containing Gal beta 1-4GlcNAc-R identified by a cytotoxic monoclonal autoantibody.
Open Access
- 1 January 1987
- journal article
- research article
- Published by Rockefeller University Press in The Journal of Experimental Medicine
- Vol. 165 (1), 124-139
- https://doi.org/10.1084/jem.165.1.124
Abstract
To investigate the autoimmune pathogenesis of spontaneously occurring diabetes mellitus in BB rats, spleen cells of newly diagnosed diabetic BB rats were fused with mouse myeloma cells. Hybridoma supernatants were screened for antibodies by indirect immunofluorescence and by 51Cr-release assays using the RINm5F rat insulinoma cell line. One clone, E5C2, produced an IgM kappa antibody that was cytotoxic for RINm5F cells, but not for other rat cell lines nor for primary rat islet cells. However, treatment of primary rat islet cells with neuraminidase exposed surface antigens and rendered the cells susceptible to complement-mediated lysis by antibody E5C2. Using immunostaining of glycolipids separated by thin-layer chromatography, hapten inhibition assays with defined carbohydrates, and Western blots, the antigens recognized by E5C2 on RINm5F cells were identified as glycoproteins with molecular weights of 60,000 and 68,000. The antibody recognizes a carbohydrate antigen containing the sequence Gal beta 1-4GlcNAc-R, which on RINm5F cells is predominantly hidden by covalently bound sialic acid. These studies raise the possibility that hidden antigenic determinants on islet cells exposed by a variety of means may be the target of autoimmune attack.This publication has 35 references indexed in Scilit:
- Autoantibodies to a 64-Kilodalton Islet Cell Protein Precede the Onset of Spontaneous Diabetes in the BB RatScience, 1984
- Monoclonal Anti-Islet Cell Autoantibodies from C57BLKsJ db/db Diabetic MiceHybridoma, 1984
- A New Technique in Quantitative Immunohematology: Solid‐Phase Kinetic Enzyme‐Linked Immunosorbent AssayVox Sanguinis, 1983
- Gangliosides of human meconium ‐ detection of a possible fetal antigenFEBS Letters, 1981
- Cytotoxic Autoantibodies to Beta Cells in the Serum of Patients with Insulin-Dependent Diabetes MellitusNew England Journal of Medicine, 1980
- A procedure for the quantitative isolation of brain gangliosidesBiochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, 1980
- Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Proceedings of the National Academy of Sciences, 1979
- Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphenylglycolurilBiochemical and Biophysical Research Communications, 1978
- Inhibition by somatostatin of insulin release from isolated pancreatic isletsFEBS Letters, 1975
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970