Cellular and tissue localization of globotriaosylceramide in Fabry disease
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The pathogenesis of Fabry disease is poorly understood. We used a variety of immunohistological techniques to localize globotriaosylceramide, the main glycolipid that accumulates in Fabry disease. Globotriaosylceramide immunoreactivity in a heterogenous pattern was present in all organs examined of a patient on long-term enzyme replacement therapy. In the brain, immmunopositivity was found only in the parahippocampal region. Globotriaosylceramide immunostaining was present in the cell membrane and cytoplasm of endothelial cells, even in the absence of lysosomal inclusions. In kidney tissue, globotriaosylceramide colocalized with lysosomal, endoplasmic reticulum, and nuclear markers. Pre- and postembedding immunogold electron microscopy of skin biopsies and untreated patient cultured skin fibroblasts confirmed the presence of globotriaosylceramide in the cell membrane, in various cytoplasmic structures, and in the nucleus. Control organ tissues and cultured fibroblasts from five unaffected subjects were uniformly negative for globotriaosylceramide by immunohistochemistry and immunogold electron microscopy. We conclude that a substantial amount of lysosomal and extralysosomal globotriaosylceramide immunoreactivity remains in cells and tissues even after years of enzyme replacement therapy in Fabry disease. These findings are crucial for the understanding of the disease mechanism and suggest the usefulness of immunostaining for globotriaosylceramide as a means to assess response to novel, specific therapies.
KeywordsGlycolipids Immunohistochemistry Electron microscopy Lysosomal disorder Lysosome
We thank the NINDS EM facility for expert technical help in the pre-embedding immunogold technique. This study was funded by the Intramural Program of the National Institute of Neurological Disorders and Stroke and the National Cancer Institute. The authors do not have financial conflict of interest that is relevant to this study. Dr. Brandon A. Wustman is employed by Amicus Therapeutics that develops pharmacological chaperones for the treatment of lysosomal diseases.
- 8.Desnick RJ, Ioannou YA, Eng CM (2001) a-Galactosidase A deficiency: Fabry disease. In: Scriver CR, Beaudet AL, Sly WS, Valle D (eds) The metabolic and molecular bases of inherited disease, 8th edn. McGraw-Hill, New York, pp 3733–3774Google Scholar
- 24.Miyamoto D, Ueno T, Takashima S, Ohta K, Miyawaki T, Suzuki T, Suzuki Y (1997) Establishment of a monoclonal antibody directed against Gb3Cer/CD77: a useful immunochemical reagent for a differentiation marker in Burkitt’s lymphoma and germinal centre B cells. Glycoconj J 14:379–388PubMedCrossRefGoogle Scholar
- 25.Miyatake T (1969) A study on glycolipids in Fabry’s disease. Jpn J Exp Med 38:135–138Google Scholar
- 27.Moore D, Scott LJC, Gladwin MT, Altarescu G, Kaneski C, Suzuki K, Pease-Fye M, Ferri R, Brady RO, Herscovitch P, Schiffmann R (2001) Regional cerebral hyper-perfusion and nitric oxide pathway dysregulation in Fabry disease: reversal by enzyme replacement therapy. Circulation 104:1506–1512PubMedGoogle Scholar
- 31.Oosterwijk E, Kalisiak A, Wakka JC, Scheinberg DA, Old LJ (1991) Monoclonal antibodies against Gal alpha 1-4Gal beta 1-4Glc (Pk, CD77) produced with a synthetic glycoconjugate as immunogen: reactivity with carbohydrates, with fresh frozen human tissues and hematopoietic tumors. Int J Cancer 48:848–854PubMedCrossRefGoogle Scholar
- 34.Pelled D, Lloyd-Evans E, Riebeling C, Jeyakumar M, Platt FM, Futerman AH (2003) Inhibition of calcium uptake via the sarco/endoplasmic reticulum Ca2+-ATPase in a mouse model of Sandhoff disease and prevention by treatment with N-butyldeoxynojirimycin. J Biol Chem 278:29496–29501PubMedCrossRefGoogle Scholar