Human IgM anti-GM1 autoantibodies modulate intracellular calcium homeostasis in neuroblastoma cells.
Increased titers of IgM anti-GM1 antibodies are present in some patients with Lower Motor Neuron Disease (LMND) or Motor Neuropathy (MN), but their pathogenic role and the mechanism of action are unclear. Previous studies have shown that the B subunit of Cholera Toxin (CT), which binds and crosslinks ganglioside GM1, modulate intracellular calcium in murine neuroblastoma cells via the activation of L-type voltage-dependent calcium channels (VGCC). Therefore, using a fluorimetric approach, we have examined the hypothesis that the pentameric IgM anti-GM1 antibodies, could similarly alter calcium concentration in N18 neuroblastoma cells. Sera with human IgM anti-GM1 antibodies were obtained from 5 patients with LMND and 2 patients with MN. Human IgG anti-GM1, IgM anti-Myelin Associated Glycoprotein (MAG), IgM anti-sulfatide antibodies and lectin peanut agglutinin (PNA), that recognizes specifically the Gal(betal-3)GalNAc epitope, were used as control sera. Direct application of either human IgM anti-GM1 antibodies or the B subunit of CT to N18 neuroblastoma cells induced a sustained influx of manganese ions, as indicated by a quench of the intracellular fura-2 fluorescence. Furthermore, the dihydropyridine L-type channel antagonists completely inhibited the manganese influx, suggesting that it is due to activation of an L-type VGCC. The magnitude of the influx was correlated with antibody titers. None of human IgG anti-GM1, IgM anti-MAG, IgM anti-sulfatide antibodies or PNA induce an ion influx, pointing to the selective participation of the pentameric IgM isotype of anti-GM1 in the modulation of L-type calcium channels opening. Given that L-type calcium channels are present on motor neurons, the modulation of L-type calcium channels by IgM GM1 antisera may have important implications in diseases such as LMND and MN.
Quattrini
A
A
San Raffaele Scientific Institute, Department of Neuroscience, Via Olgettina 60, 20132 Milan, Italy. quattrini.angelo@hsr.it
Lorenzetti
I
I
Sciorati
C
C
Corbo
M
M
Previtali
S C
SC
Feltri
M L
ML
Canal
N
N
Wrabetz
L
L
Nemni
R
R
Clementi
E
E
D000328:Adult; D001323:Autoantibodies / Q000097:blood*; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism; D002772:Cholera Toxin; D005677:G(M1) Ganglioside / Q000276:immunology*; D006706:Homeostasis / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood*; D008875:Middle Aged; D016472:Motor Neuron Disease / Q000276:immunology / Q000378:metabolism; D009447:Neuroblastoma; D009474:Neurons / Q000276:immunology* / Q000378:metabolism; D014407:Tumor Cells, Cultured
D001323:Autoantibodies; D015220:Calcium Channels; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D005677:G(M1) Ganglioside; D002772:Cholera Toxin; D002118:Calcium