Anti-ganglioside antibody internalization attenuates motor nerve terminal injury in a mouse model of acute motor axonal neuropathy
Simon N. Fewou, … , Jaap J. Plomp, Hugh J. Willison
Simon N. Fewou, … , Jaap J. Plomp, Hugh J. Willison
Published February 6, 2012
Citation Information: J Clin Invest. 2012;122(3):1037-1051. https://doi.org/10.1172/JCI59110.
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Anti-ganglioside antibody internalization attenuates motor nerve terminal injury in a mouse model of acute motor axonal neuropathy

Abstract

In the Guillain-Barré syndrome subform acute motor axonal neuropathy (AMAN), Campylobacter jejuni enteritis triggers the production of anti-ganglioside Abs (AGAbs), leading to immune-mediated injury of distal motor nerves. An important question has been whether injury to the presynaptic neuron at the neuromuscular junction is a major factor in AMAN. Although disease modeling in mice exposed to AGAbs indicates that complement-mediated necrosis occurs extensively in the presynaptic axons, evidence in humans is more limited, in comparison to the extensive injury seen at nodes of Ranvier. We considered that rapid AGAb uptake at the motor nerve terminal membrane might attenuate complement-mediated injury. We found that PC12 rat neuronal cells rapidly internalized AGAb, which were trafficked to recycling endosomes and lysosomes. Consequently, complement-mediated cytotoxicity was attenuated. Importantly, we observed the same AGAb endocytosis and protection from cytotoxicity in live mouse nerve terminals. AGAb uptake was attenuated following membrane cholesterol depletion in vitro and ex vivo, indicating that this process may be dependent upon cholesterol-enriched microdomains. In contrast, we observed minimal AGAb uptake at nodes of Ranvier, and this structure thus remained vulnerable to complement-mediated injury. These results indicate that differential endocytic processing of AGAbs by different neuronal and glial membranes might be an important modulator of site-specific injury in acute AGAb-mediated Guillain-Barré syndrome subforms and their chronic counterparts.

Authors

Simon N. Fewou, Angie Rupp, Lauren E. Nickolay, Kathryn Carrick, Kay N. Greenshields, John Pediani, Jaap J. Plomp, Hugh J. Willison

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Figure 3

Anti-GM1 is not internalized at the node of Ranvier.

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Anti-GM1 is not internalized at the node of Ranvier. (A and B) Teased sc...
(A and B) Teased sciatic nerve fibers from GD3s–/– mice labeled with anti-GM1 Ab at 4°C were further incubated at 37°C for 30–60 minutes to allow endocytosis, or studied immediately (t = 0 minutes). Anti-GM1 Ab was detected with secondary Ab in non-permeabilized tissue (A) and Triton X-100–permeabilized tissue (B) and analyzed by fluorescence microscopy. Scale bars: 20 μm. (C) Quantification of anti-GM1 Ab levels at the node of Ranvier of permeabilized and non-permeabilized tissues was conducted using ImageJ software. In tissue incubated solely at 4°C or for a further 30–60 minutes at 37°C, there was no significant difference in anti-GM1 Ab levels at the node of Ranvier in either permeabilized or non-permeabilized conditions, indicating that insignificant uptake of antibody had occurred. For quantitative data, control fibers were labeled with secondary Ab alone and showed no binding. (D) No binding to nodes of Ranvier is seen upon labeling with the anti-TNP IgG3 control Ab, followed by secondary Ab. Data are the mean of 2 independent experiment, and at least 75 nodes were assessed per condition.