Passive transfer of purified IgG from patients with amyotrophic lateral sclerosis to mice results in degeneration of motor neurons accompanied by Ca2+ enhancement.
It has been reported that immunoglobulins (IgG) in sera of patients with amyotrophic lateral sclerosis (ALS) kill cultured motoneurones (MN), but whether they also cause MN degeneration in vivo is unclear. To test this, protein-A affinity purified and dialysed IgGs were prepared from sera of 44 ALS patients without paraproteinemias, 20 healthy controls and 15 disease controls. Control and ALS-IgGs were injected intraperitoneally into groups of mice for 5 consecutive days and examined at day 8. IgG was localised immunocytochemically and spinal MN were characterised histologically and ultrastructurally and by comparative counts of Ca(2+) containing organelles revealed with oxylate-pyroantimonate histochemistry. ELISA revealed no differences in IgG concentration between ALS patients and control subjects. Immunocytochemistry showed IgG was present in MN of mice injected with control or ALS-IgG, but densitometry showed immunostaining in MN was stronger in mice injected with ALS-IgG. Compared to MN of non-injected mice, control-IgG-treated mice showed near normal MN morphology and numbers of Ca(2+)-containing organelles. Disease control IgGs evoked negligible or minor morphological changes according to disease, but normal numbers of Ca(2+) containing organelles. Ultrastructurally, about 70% of ALS-derived IgGs induced a population of MN with electron lucent cytoplasm, distended Golgi, disrupted Nissl and mitochondria (i.e., necrosis). However 30% of ALS-IgGs additionally induced electron-dense degeneration in 40% of the MN. These MN exhibited shrinkage, condensed nuclear chromatin and ill-defined nuclear membranes and resembled preliminary stages of apoptosis. We conclude that passive transfer of ALS-derived, but not control IgGs, does result in MN degeneration in the recipient mice. This appears to be associated with abnormal calcium homeostasis, but the exact target of ALS-IgG remains conjectural, and the possibilities are discussed.
Pullen
A H
AH
Sobell Department of Motor Neuroscience, Institute of Neurology, UCL, Queen Square, WC1 N 3BG, London, UK. apullen@ion.ucl.ac.uk
Demestre
M
M
Howard
R S
RS
Orrell
R W
RW
D000328:Adult; D000368:Aged; D000690:Amyotrophic Lateral Sclerosis / Q000276:immunology* / Q000473:pathology; D000818:Animals; D017209:Apoptosis; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000008:administration & dosage*; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D008875:Middle Aged; D009046:Motor Neurons / Q000187:drug effects / Q000276:immunology* / Q000473:pathology*; D009410:Nerve Degeneration; D013116:Spinal Cord / Q000187:drug effects / Q000276:immunology* / Q000473:pathology*
D007074:Immunoglobulin G