Expression of glycosylated recombinant human myelin-associated glycoprotein on a neuroblastoma cell line and its reactivity with HNK-1 but not human anti-MAG antibodies.
A correctly glycosylated myelin-associated glycoprotein (MAG) must express the carbohydrate epitope HNK-1, which is the target antigen for IgM antibodies in some patients with neuropathy. We transfected a human MAG cDNA clone into the neuroblastoma cell line SK-N-SH and verified by immunoblot the expression of the HNK-1 epitope on the recombinant molecule. By the same method and by indirect immunofluorescence we did not find any reactivity of human anti-MAG IgM antibodies with glycosylated recombinant MAG and transfected neuroblastoma cells. These findings suggest that the mere presence of the HNK-1 epitope is probably not sufficient for MAG to be recognized by human antibodies and that other factors such as the concentration or fine structure of this epitope in MAG, which mostly depend on the cellular context, may be also critical for this reactivity.
Spagnol
G
G
Centro Dino Ferrari, Institute of Clinical Neurology, Milan University, IRCCS Ospedale Maggiore Policlinico, Italy.
Doneda
P
P
Cavanna
B
B
Sgarzi
M
M
Manfredini
E
E
Allaria
S
S
Scarlato
G
G
Nobile-Orazio
E
E
D000906:Antibodies / Q000378:metabolism; D018959:CD57 Antigens / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D000939:Epitopes / Q000276:immunology / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression; D006031:Glycosylation; D006801:Humans; D015151:Immunoblotting; D018992:Myelin-Associated Glycoprotein / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D009447:Neuroblastoma / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D009474:Neurons / Q000166:cytology / Q000276:immunology; D010641:Phenotype; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012583:Schwann Cells / Q000166:cytology / Q000276:immunology; D014162:Transfection; D014407:Tumor Cells, Cultured
D000906:Antibodies; D018959:CD57 Antigens; D018076:DNA, Complementary; D000939:Epitopes; D018992:Myelin-Associated Glycoprotein; D011994:Recombinant Proteins