Florian P. Thomas

The spectrum of neurologic disease associated with anti‐GM1 antibodies

We compared anti-GM1 IgM antibody titers in patients with various neurologic diseases and in normal subjects. We found increased titers in patients with lower motor neuron disease, sensorimotor neuropathy, or motor neuropathy with or without multifocal conduction block. In patients with other diseases, titers are similar to those in normal individuals, suggesting that anti-GM1 antibody levels are not increased nonspecifically after neural injury or inflammatory diseases. Anti-GM1 antibodies in many of the patients occur as monoclonal gammopathies, predominantly of lambda light-chain type, but the antibodies are sometimes polyclonal with normal or increased serum IgM concentrations. Most of the anti-GM1 antibodies appear to react with the Gal(β1-3)GalNAc epitope which is shared with asialo-GM1 and GD1b, but in some patients the antibodies are more specific for GM1 and associated with motor neuropathy. Patients with motor or sensorimotor peripheral neuropathy or lower motor neuron disease should be tested for anti-GM1 antibodies or anti-Gal(β1-3)GalNAc antibodies, as therapeutic reduction in antibody concentrations was reported to result in clinical improvement in some patients.

Experimental autoimmune neuropathy with anti-GM1 antibodies and immunoglobulin deposits at the nodes of Ranvier

SummaryAntibodies to GM1 or Gal(β1–3)GalNAc are associated with motor or sensorimotor neuropathy and with motor neuron disease. To investigate the role of these antibodies in the neurological disorder, rabbits were immunized with GM1 or with Gal(β1–3)GalNAc-BSA, and studied serologically, electrophysiologically and pathologically. Development of antibodies to the immunizing antigens was associated with a fall in the ratio of the amplitudes of the compound muscle action potential evoked by proximal versus distal stimulation of the sciatic nerve. Pathological studies revealed mild axonal degeneration and immunoglobulin deposits at the nodes of Ranvier in peripheral nerve, resembling those reported in a patient with motor neuropathy, motor conduction block and anti-GM1 antibodies. These studies provide evidence that anti-GM1 or anti-Gal(β1–3)GalNAc antibodies cause conduction abnormalities and indicate that the antibodies may exert their effect, in part, by binding at the nodes of Ranvier in peripheral nerve.

Localization of neural epitopes that bind to IgM monoclonal autoantibodies (M-proteins) from two patients with motor neuron disease.

We investigated IgM monoclonal antibodies (M-proteins) specific for the carbohydrate epitopes Gal(beta 1-3)GalNAc and Gal(beta 1-3)GlcNAc from two patients with motor neuron disease. The M-proteins from these patients immunostain central nervous system (CNS) and peripheral nervous system (PNS) tissue from human, monkey, dog and cat at greater dilutions than tissue from rabbit, guinea pig, rat and mouse, and immunostain gray matter at greater dilutions than white matter and nerve trunks. They also bind selectively to presynaptic structures at the motor endplate region, as denervation of muscle eliminates binding. Following in vivo injection of serum into the extracellular space of the spinal cord, the M-proteins appear to bind at the surface of cells and cell processes. These studies suggest that the M-proteins might act at any one of several anatomical sites in the nervous system. This information may be helpful in selecting an animal species for further investigation of the role of M-proteins in motor neuron disease.

Tomaculous neuropathy in chromosome 1 Charcot-Marie-Tooth syndrome

We performed morphological and immunohistochemical studies on sural nerve biopsies from two members of a Charcot-Marie-Tooth type 1B family, in which a mutation of the P0 gene on chromosome 1 had been found. Biopsies showed a tomaculous neuropathy with loss of myelinated fibers and frequent small onion bulbs. Immunofluorescence with antibodies to P0 showed this protein to be present in tomaculous and non-tomaculous areas of the nyelin sheath. The severity of the myelin abnormalities suggests that in this family Charcot-marie-Tooth disease may result from a generalized disturbance of Schwann cells as a result of an abnormal P0 protein.

Human Monoclonal IgM Anti-Gal(β1-3)GaINAc Autoantibodies Bind to the Surface of Bovine Spinal Motoneurons

An IgM monoclonal autoantibody (M-protein) with anti-Gal(β1-3)GalNAc activity from a patient with lower motor neuron disease bound to the suface of motoneurons isolated from bovine spinal cord. The Gal(β1-3)GalNAc epitope is shared by the gangliosides GM1 and GD1b and by several glycoproteins in the nervous system, and binding was abolished by preabsorbing the patients serum with GM1. Antibodies specific for GM1, however, which do not bind to GaL(β1-3)GalNAc, did not bind to the motoneurons. This suggests that Gal(β1-3)GalNAc bearing glycoproteins or glycolipids other than GM1 are expressed on the surface of motoneurons, while GM1 may be absent or shielded, and that antibody binding to the cell surface might contribute to the development of the motor neuron disease.

A Monoclonal IgA in a Patient with Amyotrophic Lateral Sclerosis Reacts with Neurofilaments and Surface Antigen on Neuroblastoma Cells

A 75-year-old woman had breast carcinoma, an IgA paraprotein and autopsy-proven amyotrophic lateral sclerosis. Autopsy tissues showed immune-reactive IgA within surviving motor neurons and deposits of IgA and C3 within renal glomeruli. By indirect immunofluorescence, the patients serum contained high-titer IgA that bound to axons and to the perikarya of nerve cells in central and peripheral nervous system. The IgA paraprotein reacted with the 200 kDa, high molecular weight subunit of neurofilament protein (NFH) in Western blots of purified neurofilaments. It also reacted with dephosphorylated NFH and with NFH expressed as a fusion protein in E. coli, suggesting that the autoantibody recognized a peptide epitope. The IgA crossreacted with a surface antigen of cultured human neuroblastoma cells but mouse monoclonal antibodies to NFH did not. Absorption of the patients serum with neurofilaments eliminated IgA binding to neuroblastoma cells, indicating that the same antibodies bound to both determinants. The IgA paraprotein seems to be an autoantibody with specificity for neurofilament protein and a cell surface component of neuronal cells; the antibody may have been important in the pathogenesis of neuronal degeneration.

B cell small lymphocytic lymphoma and chronic lymphocytic leukemia with peripheral neuropathy: two cases with neuropathological findings and lymphocyte marker analysis

SummaryTwo patients with lymphoproliferative disorders developed peripheral neuropathy and neoplastic lymphocytic nerve infiltrates. One of these patients, with B cell small lymphocytic lymphoma, presented with a chronic axonal neuropathy. CD22+, CD5− cells were identified in the epineurium. The other patient with chronic lymphocytic leukemia of 3 years duration developed a mixed axonal and demyelinating neuropathy. CD22+ and CD5+ cells were observed in the endoneurium. While the cause of the neuropathy in these two cases is unknown, intraneural or systemic autoantibody production may have led to the development of disease.

Autoantigens in human neuroblastoma cells.

Neuroblastoma cells are frequently used as targets in studies of autoimmune diseases of the nervous system. We examined the human neuroblastoma cell line, LAN-5, for the presence of autoantigens that react with naturally occurring autoantibodies in human sera. Antibodies to the HNK-1 and Gal(beta 1-3)GalNAc epitopes, which have been implicated in human autoimmune neuropathy and motor neuron disease, respectively, immunostained the surface of the neuroblastoma cells, and antibodies to the 200 kDa high molecular weight neurofilament protein (NFH) immunostained the cytoplasm and cell processes. The NHK-1 and Gal(beta 1-3)GalNAc epitopes were associated with several glycoprotein bands in Western blots of the neuroblastoma cells, and the HNK-1 epitope was also shared by a glycolipid which co-migrated with 3-sulfoglucuronyl paragloboside (SGPG) from peripheral nerve, indicating that SGPG is synthesized in neuronal cells. Northern blot analysis revealed a single RNA band of 4800 bp for NFH in normal brain but two RNA species of 4800 and 3800 bp in both neuroblastoma and adrenal cells, confirming their common origin. The neuroblastoma cells appear to contain antigens that bind to naturally occurring autoantibodies in human serum and might therefore be useful for detecting and investigating the effects of anti-neuronal antibodies. The antibody populations being investigated, however, should be distinguished from other autoantibodies which might be present in the patients serum.

Antibodies to glycoconjugates in human motor neuron disease

IgM monoclonal gammopathy has been reported in some patients with motor neuron disease. The monoclonal IgMs in several of the patients bind to the carbohydrate epitope Gal (beta 1-3) GalNAc, which is shared by gangliosides GM1 and GD1b and glycoproteins in the nervous system and crossreacted with Gal (beta 1-3) GlcNAc. They also immunostain spinal cord and gray matter and presynaptic terminals of motor neurons at the neuromuscular junction. The role and mechanisms of action of these antibodies in motor neuron disease is under investigation.