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Effect of ALS IgG on Motor Neurons in Organotypic Spinal Cord Cultures

  • Bin Li (a1), Xiao-Yun Liu (a1) (a2), Zhe Li (a1), Hui Bu (a1) (a2), Meng-Meng Sun (a1), Yan-Su Guo (a1) (a2) and Chun-Yan Li (a1) (a2)...

Abstract

Objective:

Reports about the role of autoimmunity in amyotrophic lateral sclerosis (ALS) are inconsistent. The aim of this work was to investigate the effect of IgG from patients with ALS on motor neurons in a physiological-like surrounding.

Methods:

Using affinity chromatography, IgG from six ALS patients, four disease controls and five healthy subjects was purified. Organotypic spinal cord cultures, which conserve the structure of the spinal cord in a horizontal plane and are suitable for studies with long-term treatment, were used and IgG with different concentrations ranging from 0.05 mg/mL to 0.5 mg/mL was added to the culture medium. Ventral motor neuron survival was evaluated by morphology and SMI-32 immunohistochemistry staining. Lactate dehydrogenase (LDH) level in the culture medium was measured by colorimetry.

Results:

After cultures were treated with ALS IgG for three weeks, the number and morphology of motor neurons showed little change. In addition, there was no significant difference in lactate dehydrogenase release between cultures treated with medium alone, normal control IgG, disease control IgG or ALS IgG.

Conclusions:

The results indicate that IgG from these ALS patients was insufficient per se to induce motor neuron death in Organotypic slice cultures. However, this does not preclude the possibility that other changes may have occurred in the motor neurons. This work offered a new model to evaluate the role of IgG in the pathogenesis of ALS. Organotypic cultures contribute to study of the impact of IgG on motor neurons by mimicking physiological conditions.

<span class='bold'>RÉSUMÉ:</span> <span class='bold'> <span class='italic'>Objectif:</span> </span>

Les données sur le röle de l'autoimmunité dans la genèse de la sclérose latérale amyotrophique (SLA) sont contradictoires. Le but de cette étude était de déterminer quels sont les effets d'IgG provenant de patients atteints de SLA sur des neurones moteurs dans un milieu simulant le milieu physiologique.

<span class='bold'> <span class='italic'>Méthodes:</span> </span>

Des IgG provenant de six patients atteints de SLA, de quatre patients atteints d’autres maladies et de cinq sujets sains ont été purifiées par chromatographie d’affinité. Des cultures organotypiques de moelle épinière qui conservent la structure de la moelle épinière sur un plan horizontal et qui sont adéquates pour des études à long terme, ont été utilisées. Des IgG à différentes concentrations, allant de 0,05 mg/mL à 0,5 mg/mL, ont été ajoutées au milieu de culture. La survie des neurones moteurs antérieurs a été évaluée par examen morphologique et par coloration immunohistochimique au SMI–32. Le niveau de déshydrogénase lactique (DHL) dans le milieu de culture a été mesuré par colorimétrie.

<span class='bold'> <span class='italic'>Résultats:</span> </span>

: Le nombre et la morphologie des neurones moteurs étaient pratiquement inchangés après trois semaines de traitement des cultures par des IgG provenant de patients atteints de SLA. De plus, il n'existait pas de différence significative dans la libération de DHL entre les cellules cultivées dans le milieu de culture sans ajout, le milieu de culture contenant des IgG de sujets témoins normaux, le milieu de culture contenant des IgG de patients atteints d’autres maladies et le milieu de culture contenant des IgG de patients atteints de SLA.

<span class='bold'> <span class='italic'>Conclusion:</span> </span>

Ces résultats indiquent que les IgG des patients atteints de SLA n’étaient pas suffisantes pour induire la mort de neurones moteurs en cultures organotypiques de coupes de tissus. Cependant, ceci n’exclut pas la possibilité que d’autres changements étaient présents dans les neurones moteurs. Cette étude représente un nouveau modèle pour évaluer le röle des IgG dans la pathogenèse de la SLA. Les cultures organotypiques contribuent à l’étude de l’impact des IgG sur les neurones moteurs en imitant les conditions physiologiques.

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Copyright

Corresponding author

Department of Neurology, the Second Hospital of Hebei Medical University, Institute of Cardiocerebrovascular Disease, Hepingxi Road 215, Shijiazhuang, Hebei, People’s Republic of China, 050000.

References

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