Summary
Epstein-Barr virus-transformed lymphoid cell lines from normal individuals and from patients with Niemann-Pick disease types A, B or C were subjected to various culture conditions in order to study the source of the characteristic lysosomal storage of sphingomyelin observed in the tissues of Niemann-Pick patients. The culture medium was supplemented with a serum substitute devoid of lipoproteins or with one of the following lipid sources: fetal calf serum, human low-density lipoprotein (LDL), or human high-density lipoprotein (HDL). Storage of sphingomyelin was demonstrated under all tested culture conditions in cells deficient in acid sphingomyelinase (Niemann-Pick disease types A and B). In contrast, the sphingomyelin concentration in the lymphoid cell line from a Niemann-Pick type C patient (not deficient in sphingomyelinase) was normal. After more than 30 days in a medium devoid of sphingomyelin, the Niemann-Pick types A and B lymphoid cell lines showed accumulation of sphingomyelin about twice control. The concentration was higher when cells were grown in a medium supplemented with lipids, particularly human LDL or HDL. These results are consistent with the hypothesis that both exogenous and endogenous sphingomyelins participate in the lysosomal storage observed in lymphoid cell lines from patients with Niemann-Pick disease types A and B.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ames, B. N. Assay of inorganic phosphate, total phosphate and phosphatases.Methods Enzymol. 8 (1966) 115–118
Beaudet, A. L. and Manschreck, A. A. Metabolism of sphingomyelin by intact cultured fibroblasts: differentiation of Niemann-Pick disease types A and B.Biochem. Biophys. Res. Commun. 105 (1982) 14–19
Bes, J. C., Salvayre, R., Levade, T., Caratero, C. and Planel, H. Ultrastructural investigations on two lymphoid cell lines from Niemann-Pick disease type B.Biol. Cell 50 (1984) 299–302
Brady, R. O. Sphingomyelin lipidoses: Niemann-Pick disease. In Stanbury, J. B., Wyngaarden, J. B., Fredrickson, D. S., Goldstein, J. L. and Brown, M. S. (eds.)The Metabolic Basis of Inherited Disease, McGraw-Hill, New York, 1983, pp. 831–841
Elleder, M. and Jirasek, A. Niemann-Pick disease.Acta Universitatis Carolinae Medica 29 (1983) 259–267
Fishman, P. H., Bradley, R. M., Brown, M. S., Faust, J. R. and Goldstein, J. L. Similar content of phospholipids and gangliosides in normal and homozygous familial hypercholesterolaemia fibroblasts.J. Lipid Res. 19 (1978) 304–308
Folch, J., Lees, M. and Sloane-Stanley, G. H. A simple method for the isolation and purification of total lipids from animal tissues.J. Biol. Chem. 226 (1957) 497–509
Forte, T. M., Bell-Quint, J. J. and Cheng, F. Lipoproteins of fetal and newborn calves and adult steer: a study of developmental changes.Lipids 16 (1981) 240–245
Goldstein, J. L. and Brown, M. S. Binding and degradation of low-density lipoproteins by cultured human fibroblasts.J. Biol. Chem. 249 (1974) 5153–5162
Hartree, E. F. Determination of protein: a modification of the Lowry method that gives a linear photometric response.Anal. Biochem. 48 (1972) 422–427
Ho, Y. K., Brown, M. S., Kayden, H. J. and Goldstein, J. L. Binding, internalization and hydrolysis of low-density lipoprotein in long-term lymphoid cell lines from a normal subject and a patient with homozygous familial hypercholesterolaemia.J. Exp. Med. 144 (1976) 444–455
Jackson, R. L., Morrisett, J. D. and Gotto, A. M. Lipoprotein structure and metabolism.Physiol. Rev. 56 (1976) 259–316
Kayden, H. J., Hatam, L. and Beratis, N. G. Regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity and the esterification of cholesterol in human long-term lymphoid cell lines.Biochemistry 15 (1976) 521–528
Kudoh, T., Velkoff, M. A. and Wenger, D. A. Uptake and metabolism of radioactively labelled sphingomyelin in cultured skin fibroblasts from controls and patients with Niemann-Pick disease and other lysosomal storage diseases.Biochim. Biophys. Acta 754 (1983) 82–92
Leikin, A. I., Mihovilovic, M. and Scanu, A. M. High-density lipoproteins influence cholesterol homeostasis in cultured virus-transformed human lymphoblastoid cells.J. Biol. Chem. 257 (1982) 14280–14287
Levade, T., Salvayre, R., Lenoir, G. and Douste-Blazy, L. Sphingomyelinase and non-specific phosphodiesterase activities in Epstein-Barr virus-transformed lymphoid cell lines from Niemann-Pick disease A, B and C.Biochim. Biophys. Acta 793 (1984) 321–324
Levade, T., Salvayre, R. and Douste-Blazy, L. Molecular forms of sphingomyelinase and non-specific phosphodiesterases in Epstein-Barr virus-transformed lymphoid cell lines from Niemann-Pick disease A and B.Eur. J. Biochem. 149 (1985a) 405–409
Levade, T., Salvayre, R., Bes, J. C., Maret, A. and Douste-Blazy, L. Biochemical and ultrastructural findings in a lymphoid cell line from Niemann-Pick disease type A.Biol. Cell 55 (1985b) 143–146
Levade, T., Salvayre, R., Bes, J. C., Nezri, M. and Douste-Blazy, L. New tools for the study of Niemann-Pick disease: analogues of natural substrate and Epstein-Barr virus-transformed lymphoid cell lines.Pediatr. Res. 19 (1985c) 153–157
Levade, T., Maret, A., Salvayre, R., Livni, N., Rogalle, P. and Douste-Blazy, L. Biochemical and ultrastructural studies on an Epstein-Barr virus-transformed lymphoid cell line from a Niemann-Pick disease type C patient.Biochim. Biophys. Acta. 877 (1986a) 415–422
Levade, T., Salvayre, R. and Douste-Blazy, L. Sphingomyelinases and Niemann-Pick disease.J. Clin. Chem. Clin. Biochem. 24 (1986b) 205–220
Schumaker, V. N. and Puppione, D. L. Sequential flotation ultracentrifugation.Methods Enzymol. 128 (1986) 155–170
Spence, M. W., Clarke, J. T. R. and Cook, H. W. Pathways of sphingomyelin metabolism in cultured fibroblasts from normal and sphingomyelin lipidosis subjects.J. Biol. Chem. 258 (1983) 8595–8600
Sutrina, S. L. and Chen, W. W. Lysosomal involvement in cellular turnover of plasma membrane sphingomyelin.Biochim. Biophys. Acta 793 (1984) 169–179
Vanier, M. T., Rousson, R., Garcia, I., Bailloud, G., Juge, M. C., Revol, A. and Louisot, P. Biochemical studies in Niemann-Pick disease. III.Clin. Genet. 27 (1985) 20–32
Wenger, D. A. Niemann-Pick disease. In Glew, R. H. and Peters, S. P. (eds.)Practical Enzymology of the Sphingolipidoses, Alan R. Liss, New York, 1977, pp. 39–70
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Levade, T., Salvayre, R., Maret, A. et al. Evidence for both endogenous and exogenous sources of the sphingomyelin storage in lymphoid cell lines from patients with Niemann-Pick disease types A and B. J Inherit Metab Dis 11, 151–157 (1988). https://doi.org/10.1007/BF01799864
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01799864