Anti-ganglioside antibodies alter presynaptic release and calcium influx.

Acute motor axonal neuropathy (AMAN) variant of Guillain-Barré syndrome is often associated with IgG anti-GM1 and -GD1a antibodies. The pathophysiological basis of antibody-mediated selective motor nerve dysfunction remains unclear. We investigated the effects of IgG anti-GM1 and -GD1a monoclonal antibodies (mAbs) on neuromuscular transmission and calcium influx in hemidiaphragm preparations and in cultured neurons, respectively, to elucidate mechanisms of Ab-mediated muscle weakness. Anti-GM1 and -GD1a mAbs depressed evoked quantal release to a significant yet different extent, without affecting postsynaptic currents. At equivalent concentrations, anti-GD1b, -GT1b, or sham mAbs did not affect neuromuscular transmission. At fourfold higher concentration, an anti-GD1b mAb (specificity described in immune sensory neuropathies) induced completely reversible blockade. In neuronal cultures, anti-GM1 and -GD1a mAbs significantly reduced depolarization-induced calcium influx. In conclusion, different anti-ganglioside mAbs induce distinct effects on presynaptic transmitter release by reducing calcium influx, suggesting that this is one mechanism of antibody-mediated muscle weakness in AMAN.

Presynaptic effects of immunoglobulin G from patients with Lambert-Eaton myasthenic syndrome: their neutralization by intravenous immunoglobulins.

Intravenous immunoglobulin (IVIg) treatment improves muscle strength in Lambert-Eaton myasthenic syndrome (LEMS), but its specific mode of action is unknown. We have delineated its mode of action on neuromuscular blocking properties of LEMS IgG. The effect of sera and purified IgG from six patients with LEMS on evoked quantal release was investigated after direct application to the motor nerve terminal by the perfused macro-patch-clamp electrode in mouse hemidiaphragms. The effect of LEMS IgG was analyzed alone and after coincubation with different concentrations of IVIg or its Fab fragments. All LEMS sera and purified LEMS IgG fractions taken before IVIg treatment inhibited evoked quantal release in a dose-dependent manner. When LEMS IgG was coincubated with a therapeutic IVIg preparation, presynaptic inhibitory activity of LEMS IgG was diminished in a dose-dependent fashion. Monovalent Fab fragments were as effective in neutralizing the activity of LEMS IgG as whole IVIg. These direct neutralizing effects of IVIg may explain its therapeutic efficacy.

Differential blocking effects of the monoclonal anti-GQ1b IgM antibody and alpha-latrotoxin in the absence of complement at the mouse neuromuscular junction.

Numerous in vitro electrophysiological studies found different effects of anti-ganglioside antibodies on neuromuscular transmission. Since a complement-dependent, latrotoxin-like effect has been described for the mouse monoclonal anti-GQ1b IgM antibody (termed CGM3), we here investigated the effect of CGM3 and alpha-latrotoxin by means of the perfused macro-patch clamp electrode in mice hemidiaphragms in the absence of complement. The CGM3 mab depressed evoked quantal release dose-dependently, whereas the rate of spontaneous releases and the amplitude of postsynaptic currents was not significantly affected. Alpha-latrotoxin induced an increase in spontaneous releases followed by a blockade of evoked quantal release, this was not altered by CGM3. The complement-independent presynaptic block by CGM3 was different from the action of alpha-latrotoxin.

Intravenous immunoglobulins neutralize blocking antibodies in Guillain-Barré syndrome.

Intravenous immunoglobulin (IVIg) treatment ameliorates the course of Guillain-Barré syndrome (GBS), but its specific mode of action is unknown. We attempted to delineate the effect of IVIg on neuromuscular blocking antibodies in GBS. A total of seven GBS serum samples were examined for blocking antibodies and the effect of IVIg with a macro-patch-clamp technique in mouse hemidiaphragms. First, serum was tested before and after treatment with IVIg. Second, we investigated with coincubation experiments whether the IVIg was capable of neutralizing neuromuscular blocking antibodies in GBS serum or affinity-purified immunoglobulin G (IgG) fractions. Finally, the mechanism of the neutralizing effect was studied by the coincubation of active blocking GBS IgG with Fab and Fc fragments prepared from IVIg. All GBS sera (two adults and two children) and GBS IgG fractions (three adults) taken before treatment with IVIg blocked evoked quantal release by approximately 90%. Blocking activity was markedly reduced in sera obtained after treatment with IVIg. Coincubation of the pretreatment blocking serum with the posttreatment serum, or with the IVIg preparation used for treatment, reduced the blocking activity of the pretreatment GBS serum. When GBS IgG was coincubated with IVIg, the blocking activity of GBS IgG was diminished dose-dependently. Monovalent and divalent Fab fragments prepared from the IVIg were as effective as whole IVIg, but Fc fragments were ineffective. Therapeutic IVIg is capable of neutralizing neuromuscular blocking antibodies in GBS by a dose-dependent, antibody-mediated mechanism. This may, in part, explain its therapeutic efficacy.