Y. Graus

Characterization of anti‐acetylcholine receptor (AChR) antibodies from mice differing in susceptibility for experimental autoimmune myasthenia gravis (EAMG)

In the murine model for EAMG we investigated the relation between disease susceptibility and fine specificity of anti‐AChR antibodies obtained from high susceptible C57BI/6 and low susceptible BALB/c mice after immunization with Torpedo acetylcholinc receptor (tAChR). Anti‐AChR MoAbs with fine specificity for the main immunogenic region (MIR), the α‐bungarotoxin (α‐BT)/ acetylcholine binding sites and other extra‐ and intracellular epitopes were isolated from both mouse strains. In total, nine out of 38 MoAbs obtained from C57BI/6 mice were directed against extracellular epitopes on mouse AChR in contrast to only one out of 27 MoAbs from BALB/c mice. A difference in antibody repertoire may underlie the difference in pathogenic response observed between these mouse strains. These results indicate that strain‐specific differences in disease susceptibility in murine EAMG may be related to differences in the available repertoire of potential pathogenic antibodies.

Age- and sex-related resistance to chronic experimental autoimmune myasthenia gravis (EAMG) in Brown Norway rats.

The influence of age and sex on the induction of chronic EAMG was analysed. Aged male rats, immunized with Torpedo acetylcholine receptor (tAChR), showed no clinical signs of disease or AChR loss. Immunization of young male Brown Norway (BN) rats resulted in both clinical signs of disease and 65% AChR loss. In contrast, both young and aged female BN rats showed comparable AChR loss (58% and 50%, respectively), although aged female rats did not develop clinical signs of disease. Differences in antibody titres, isotype distribution, fine specificity or complement activation could not account for the observed resistance. These results suggest that resistance against EAMG in aged rats is due to resistance of the AChR against antibody‐mediated degradation, or to mechanisms able to compensate for AChR loss.

MACROPHAGE INFILTRATION AT THE NEUROMUSCULAR JUNCTION DOES NOT CONTRIBUTE TO ACHR LOSS AND AGE-RELATED RESISTANCE TO EAMG

Aged rats resistant to acetylcholine receptor loss in passive transfer experimental autoimmune myasthenia gravis (EAMG) do not reveal infiltrating macrophages at the neuromuscular junction (NMJ) as observed in susceptible rats. It was investigated whether this age-related resistance is due to impaired macrophage function in these aged rats. Reconstitution of aged rats with bone marrow from young donors did not lead to macrophage infiltration, nor did it abolish resistance to EAMG. Subsequently, it was investigated whether macrophages are a primary cause of acetylcholine receptor (AChR) loss in EAMG or are attracted to the NMJ secondary to tissue damage. In lethally irradiated young susceptible rats infiltrating macrophages were absent from the NMJ. However, similar AChR losses were observed in irradiated and non-irradiated rats. These results suggest that macrophages do not contribute to acetylcholine receptor loss in the effector phase of passive transfer EAMG and that age related resistance to passive transfer EAMG is not primarily determined by the absence of infiltrating macrophages.

Age related resistance to experimental autoimmune myasthenia gravis in rats

The influence of age on the induction of experimental autoimmune myasthenia gravis (EAMG) was investigated. Immunization with acetylcholine receptor (AChR) or injection of varying amounts of anti-AChR mAb 35 into young adult (10-12 wk) BN rats induced severe signs of EAMG including weight loss and decrement of muscle action potential, whereas aged BN rats (120-130 wk) did not show any clinical signs of EAMG. Serum anti-AChr mAb titers were not significantly different in young and aged rats up to 24 h after administration of mAb. No significant AChR loss was demonstrated in aged rats, whereas similarly treated young rats showed extensive AChR loss. In contrast to young rats, no degradation of the postsynaptic membrane could be demonstrated by electron microscopy in aged rats. C component C3 and C5b-9 membrane attack complex could be demonstrated at the neuromuscular junction in both young and aged mAb-treated rats. However, infiltrating macrophages and necrotic muscle fibers were seen only in young rats. These results suggest that the postsynaptic membrane in aged rats is resistant to autoantibody attack. AChR degradation by antigenic modulation may be less efficient in aged rats as a result of altered AChR density and distribution or rigidity of the postsynaptic membrane. Age-related resistance in the EAMG model can provide more information about the factors that determine the severity of myasthenia gravis. Manipulation of AChR density or lipid composition of the postsynaptic membrane may be of therapeutic interest in myasthenia gravis.