Lymphocytes transfer only the lysosomal form of alpha-D-mannosidase during cell-to-cell contact.
We have examined the changes in the activities of the different types of alpha-D-mannosidase when fibroblasts from patients deficient in the lysosomal form of the enzyme are cultured together with normal lymphocytes. Our results show that whereas the mannosidosis cells acquired high levels of this enzyme, the activities of both the Golgi and the endoplasmic reticulum forms of alpha-D-mannosidase remained the same as in the fibroblasts cultured alone in the absence of lymphocytes. The increase in the activity of the lysosomal enzyme in the cocultured fibroblasts was not affected by the presence of mannose 6-phosphate or alpha-methyl mannoside, inhibitors of receptor- and lectin-mediated uptake of lysosomal enzymes, respectively, but it did require cell-to-cell contact. Ion-exchange HPLC and electrophoresis in polyacrylamide gradient gels showed that the acquired enzyme had the same elution profile and molecular size as the lysosomal form of the enzyme present in the lymphocytes. Immunoprecipitation studies using antibody specific for the lymphocyte type of lysosomal alpha-D-mannosidase confirmed that the increased activity in the cocultured mannosidosis cells resulted from the acquisition of the lymphocyte enzyme. Cytochemical examination revealed, however, that the transferred lymphocyte enzyme was localized in cytoplasmic organelles in the peripheral regions of the recipient fibroblasts. These results show that lymphocytes transfer only the lysosomal form of alpha-D-mannosidase during cell-to-cell contact with mannosidosis cells.
Abraham
D
D
Kennedy Institute of Rheumatology, Bute Gardens, London, United Kingdom.
Muir
H
H
Winchester
B
B
Olsen
I
I
D000818:Animals; D001692:Biological Transport; D002448:Cell Adhesion; D002450:Cell Communication; D002478:Cells, Cultured; D011232:Chemical Precipitation; D005347:Fibroblasts / Q000201:enzymology; D006651:Histocytochemistry; D006801:Humans; D066298:In Vitro Techniques; D007526:Isoelectric Point; D008247:Lysosomes / Q000166:cytology / Q000201:enzymology*; D008361:Mannosidases / Q000378:metabolism*; D051379:Mice; D008970:Molecular Weight; D008363:alpha-Mannosidosis / Q000201:enzymology
D008361:Mannosidases