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Antiganglioside antibodies: When, which and for what
Abstract
We report our experience in the study of antiganglioside antibodies and define their clinical value establishing associations between clinical syndromes and immunological findings.
We analysed 275 sera: Guillain-Barré syndrome (GBS) (78), Miller-Fisher syndrome (MFS) (37), chronic inflammatory demyelinating polyneuroapthy (CIDP) (17), multifocal motor neuropathy (NMM) (42), chronic axonal mixed polyneuropathy (PNP) (54), amyotrophic lateral sclerosis (ALS) (28) and lower motor neuron disease (LMND) (17). We have studied the presence of IgG and IgM antibodies to 9 gangliosides using ELISA and TLC.
We have detected anti-GQ1b antibodies in 36/37 (97,3%) of patients with MFS, being undetectable after 4 weeks in 83%. A 34 % (26/78) of patients with GBS were positive for several antiganglioside specificities being GalGalNAc the most frequent (54%). Two out of three sera positive for GD1a corresponded to axonal Guillain-Barré. IgM class anti-GM1 antibodies were positive in 10/12 patients with MMN, while only a 3-9% of patients with ALS, CIDP, PNP and LMND presented antiganglioside antibodies.
Analysis of anti-GQ1b antibodies confirms the diagnosis of MFS, excluding other acute brainstem pathologies and, in this study, detection of anti-GD1a antibodies indicates axonal damage in GBS and suggest a worse prognosis. IgM anti-GM1 antibodies are only found in MMN. These findings confirm a disease specific correlation between specific neuropathies and antiganglioside antibodies clinically useful.
... Por otro lado, su enriquecimiento en tejido neural llevó a asociar a estos GSL con el funcionamiento del sistema nervioso (Ho et al. 1997a;Sheikh et al. 1998) y consecuentemente con trastornos neurológicos de origen metabólico (enfermedad de Tay-Sachs) y enfermedades neurodegenerativas (Sandhoff y Harzer 2013;Ariga 2014;Aureli et al. 2015). Adicionalmente, en diversas neuropatías de origen autoinmune, los gangliósidos han sido implicados adoptando el rol de antígenos (auto-antígenos) (Ilyas et al. 1988;Gallardo et al. 2001;Ariga 2014). ...
... Desde la década de los 80, la presencia de anticuerpos anti-gangliósidos de especificidades bien definidas ha sido correlacionada con diversos síndromes clínicos (Ilyas et al. 1985a;Ilyas et al. 1985b;Freddo et al. 1986;Gallardo et al. 2001;Ariga 2014). Si bien en el suero de individuos sanos se detectan anticuerpos anti-gangliósido, éstos poseen títulos y afinidad bajos ), a diferencia de los pacientes con SGB que poseen anticuerpos con afinidades elevadas. ...
Gangliosides are abundant glycans in neural tissue. They have been proposed as one of the major antigenic targets in Guillain-Barré syndrome (SGB). Although the etiology of this autoimmune disease is unknown, it is believed that the immune response elaborated to a pathogen could cross-react with peripheral nerve gangliosides. Rabbits immunized with bovine brain gangliosides (BBG) and keyhole limpet hemocyanin (KLH) like carrier protein develops a neuropathy clinically similar to SGB. However the disease does not occur if methylated bovine serum albumin (BSAm) is used instead KLH.
In order to characterize the development of humoral immune response in this animal model, groups of animals were immunized with different immunogens. A group was immunized with KLH/BBG and another one with BSAm/BBG. To evaluate the carrier function of KLH in the experimental model also two other groups were immunized with an alternative immunization protocol (BSAm/BBG+KLH and KLH+BBG) in which KLH was administered in a separate emulsion. On the other hand, two control groups immunized only with gangliosides or only with KLH were also used. Clinical signs and the immune response generated (title, and isotype specificity of the anti- gangliosides antibodies) were evaluated in all groups.
All the animals immunized with KLH/BBG showed clinical signs of neuropathy and IgG antibodies were detected that recognize simple (GA1) and complex (GM1 and GD1b) structure gangliosides. These antibodies were partially blocked by pre-incubation with soluble KLH indicating cross-reactivity. The percentage of cross-reactivity with KLH was modified over time (change in antibodies specificity). The reduction in the percentage of cross-reactive antibodies generally coincides with the onset of clinical signs. No animals of the control groups got sick (KLH or BBG groups). Only in rabbits immunized with KLH was detected a slight reactivity of IgG isotype antibodies that recognize GA1, and was completely blocked by preincubation with soluble KLH. Using the peanut agglutinin (PNA) was confirmed in KLH, the existence of a determinant also present in GA1/GM1/GD1b (Galβ1-3GalNAc). While the BSAm/BBG group did not get sick, when we also immunized this group with KLH separately (BSAm/BBG+KLH) the situation is reversed and neuropathy is generated. Nevertheless separately immunization with KLH and BBG (KLH+BBG) did not produce clinical signs of disease. Finally among the animals that developed neuropathy, those with an early onset had a higher percentage of high affinity anti-GA1 antibodies at pre-immune serum compared with those with a late onset.
These results suggest that the presence of KLH in the immunization is required for development the disease. The antibodies begin recognizing the terminal sugars shared between KLH-gangliosides and then expand the identified region. Consequently it occur a change of antibodies specificity to gangliosides, accompanied by the onset of neuropathy. Although gangliosides do not possess the ability to generate an IgG isotype response, it may redirect the anti-GA1 response produced by KLH against gangliosides with more complex structures. Fine specificity and affinity of the anti-gangliosides antibodies are key in the onset of neuropathy, in fact a high percentage of anti-GA1 high affinity pre-immune antibodies was associated with early onset of the disease.
Two main conclusions are established. First, KLH would have two fundamental roles, carrier and specific immune stimulant. Second, the susceptibility factor to develop the neuropathy may be related to the affinity of anti-GA1 NOAbs present in the pre-immune serum. To our knowledge, this work is the first not only to describe in detail the immune response generated during the development of neuropathy in an experimental model but also to establish a possible host susceptibility factor.
... The investigation of other ganglioside antibodies of alpha series, GM2, GM3, GD1a, GT1a is our next target of research in Parkinson's disease. Moreover we focused on IgM antibodies against GM1, GD1b and GQ1b since they are the most widely ganglioside antibodies studied with ELISA [46][47][48][49][50]. In addition we attempted to correlate the results with the clinical parameters, for figuring out an existing possible correlation, eventually. ...
... According to other authors, Aβ-peptide (1-42) binds tightly to GM1, compared to the lower binding affinity for GD1b and GQ1b [57,58]. Increased anti-GM1 and anti-GD1b IgM were also detected in elderly patients suffered from dementia [49,45]. ...
Background: Increasing evidence suggests that gangliosides act as important mediators in both demyelination and remyelination.
Objective: The purpose of the present study was to investigate the presence of IgM antibodies against GM1, GD1b and GQ1b gangliosides in the sera of in patients who suffered from Parkinson’s disease in correlation with the clinical parameters.
Method: The present research is based on the investigation of 44 patients (63.7-73.3 years) for anti-ganglioside antibodies and 44 healthy age- matched individuals, as normal controls, using Enzyme-Linked Immuno-Sorbent Assays.
Results: The patients revealed increased levels of the tested antibodies, compared to normal controls (p=0.0005). A correlation between IgM anti-GM1 and the level of cognitive impairment (Minimental State Examination, p=0.003; Unified Parkinson’s Disease Rating Scale I, p=0.013) was also noticed. Conclusion: A peripheral neuroimmune response may occur in patients who suffer from Parkinson’s disease especially those with cognitive impairment. Further investigation is needed to establish a direct connection between that immune response and disease pathophysiology.
... Der Nachweis von Anti-Gangliosid-Antikörpern stellt kein typisches Charakteristikum bei der klassischen CIDP dar. Während bei der MMN, einem Subtyp der CIDP, bis zu 80% der Patienten Antikörper gegen GM1 aufweisen (Gallardo et al., 2001;Hughes et al., 2006), besitzen nur zwischen 0-23% der Patienten mit klassischer CIDP IgG-Antikörper gegen GM1 (Simone et al., 1993;van Schaik et al., 1994;Yuki et al., 1996;Gallardo et al., 2001). Ganglioside werden mit vielen physiologisch bedeutenden Funktionen in Verbindung gebracht, die durch die Bindung von pathogenen Antikörpern beeinträchtigt werden können. ...
... Der Nachweis von Anti-Gangliosid-Antikörpern stellt kein typisches Charakteristikum bei der klassischen CIDP dar. Während bei der MMN, einem Subtyp der CIDP, bis zu 80% der Patienten Antikörper gegen GM1 aufweisen (Gallardo et al., 2001;Hughes et al., 2006), besitzen nur zwischen 0-23% der Patienten mit klassischer CIDP IgG-Antikörper gegen GM1 (Simone et al., 1993;van Schaik et al., 1994;Yuki et al., 1996;Gallardo et al., 2001). Ganglioside werden mit vielen physiologisch bedeutenden Funktionen in Verbindung gebracht, die durch die Bindung von pathogenen Antikörpern beeinträchtigt werden können. ...
... это приводит к изменению патофизиологического континуума от раннего обратимого нарушения проводимости до дегенерации аксонов [14]. Эти данные подтверждают, что возрастные характеристики коррелируют с специфической невропатией и антитела к ганглиозидам могут быть клинически полезны [15]. ...
Nowadays, the percentage of elderly people in society grows. Good nutrition and medical care help older people to have a normal life over 80 to 90 years. In the last ten years it is of critical importance to establish the clinical significance of serum IgG anti-GD1a and anti-GM1 ganglioside antibodies as potential biomarkers for neuronal damage in neurodegenerative diseases and immune-mediated neuropathies and demyelination. In the current study, the diagnostic values of IgG anti-GD1a and anti-GM1 antibodies were determined by the ELISA method in serum samples of 18 elderly patients (71-91 years). Significantly elevated serum IgG anti-GD1a and anti-GM1 antibodies titers were detected only in patients over 80 years. These data suggest that the immune-mediated neuropathies, neurodegeneration and demyelination in healthy elderly occur after 80 years old. Therefore, IgG anti-GD1a and anti-GM1 antibodies can serve as biomarkers, showing the nervous system dysfunction.
... 11 GM1 are the main myelin ganglioside and the high titer of IgG anti-GM1 antibodies is correlated with the demyelination. 12 Serum gangliosides GM3 could be used as biomarkers of BBB destruction. 13 The aim of the present investigation is to value the serum titers of the IgG anti-GD1a, anti-GM1 and anti-GM3 antibodies in aging rats. ...
Objectives: We measure IgG gangliosides antibodies titers in rats sera. Background: The complex of anti-gangliosides antibodies may be useful diagnostic and prognostic markers of the neurodegeneration (GD1a), demyelination (GM1) and correlated with the loss of integrity of the blood brain barrier (GM3). Methods: The values of IgG anti-GD1a, anti-GM1 and anti-GM3 antibodies titers were detected by ELISA in the rats (1, 13, 18, 22, 30, 33 months of age). Results: Significantly elevated serum IgG anti-GD1a, anti-GM1 and anti-GM3 antibodies titers were detected in rats at age only over 18 months. At 18 and 22 monthly rats significant increase in titer of antibodies against GD1a was shown. At 30 and 33 monthly rats significant increase in the serum titer of antibodies against GM1 and GM3 was shown. Conclusions: Our results suggest that it is possible to use the serum IgG antibodies as biomarkers for neurodegeneration, demyelination and the damage of the integrity of the blood brain barrier. The value of the titer of IgG antibodies against GD1a, GM1 and GM3 ganglioside showed the presence of immunological mediated neurodegeneration, concomitant demyelination, growing with age, and impaired the integrity of the blood brain barrier. The observed changes are evidence of age-related neurodegenerative changes.
Resumen
Las neuropatías periféricas son una patología neurológica frecuente y compleja debida a la gran variedad de causas que pueden producir un mismo patrón de afectación clínica. Se caracterizan por síntomas motores, sensitivos y autonómicos que pueden seguir varios patrones de distribución (mononeuropatía, mononeuropatía múltiple, polirradiculoneuropatía y polineuropatía distal simétrica). Tras la tipificación clínica, se solicitarán diversas pruebas complementarias (estudio electrodiagnóstico, pruebas de laboratorio, punción lumbar, biopsia de nervio periférico o de piel), encaminadas a filiar la posible etiología del patrón clínico que presenta el paciente. Es importante llegar a un diagnóstico etiológico en aquellas neuropatías susceptibles de tratamiento. Las radiculopatías son otra patología frecuente del sistema nervioso periférico, generalmente debidas a la compresión mecánica de la raíz por causas degenerativas (espondiloartrosis, hernia discal). Las lesiones del plexo braquial o lumbosacro son menos frecuentes, y pueden obedecer a traumatismos, lesiones postradioterapia o por infiltración metastásica, entre otras.
A number of autoantibodies that induce inflam-mation on autoimmune peripheral neuropathies have been described. We review the techniques to measure autoantibodies and assess the usefulness of antibody assays in acquired acute demyelinating neuropathies such as Guillain-Barré syndrome (GBS) and chronic acquired demyelinating neuro-pathies including CIDP, multifocal motor neuro-pathy and MGUS neuropathy. In acute acquired demyelinating neuropathies associations of clinical characteristics and specific infections and the presence of anti-ganglioside antibodies have been found. Since diagnostic criteria have been avail-able to subclassify Guillain-Barré syndrome in dif-ferent clinical variants including clinical, electro-physiological, pathological and immunological findings, several varieties have been described: pure motor forms, sensory forms, primary axonal and primary demyelinating varieties. However, further studies are necessary to validate the use-fulness of this subclassification with respect to treatment and prognosis.This is particularly impor-tant if subgrouping of GBS patients may lead to more individualised treatment. It has been sug-gested that for the IgG anti-GM1-positive sub-group of GBS patients, IVIg therapy may be the more efficacious treatment than plasmapheresis. The spectrum of chronic acquired demyelinating polyneuropathies cover different entities that have recently been categorised in three major groups: chronic inflammatory demyelinating polyneuro-pathy (CIDP), multifocal motor neuropathy (MMN) and MGUS neuropathy. Though the majority of CIDP patients respond well to immunotherapy, no constant autoantibody activity has been reported. In MMN serum antibodies, mostly IgM to the ganglioside GM1 or less frequently to asialo-GM1, GD1a or GM2 have been reported by ELISA in a variable proportion of MMN patients with a prevalence for anti-GM1 IgM in most large series, ranging from 30 to 60%. The reasons for these dis-crepant figures are still unclear but may be related to differences in the ELISA procedure or in the controls used to establish normal reference values. Antibodies to the myelin-associated glycoprotein (MAG) are detected in 50 to 60% of patients with neuropathy and immunoglobulin M (IgM) mono-clonal gammopathy. Most patients have a slowly progressive, sensory or sensorimotor, demyelinat-ing polyneuropathy. A causal relation between anti-MAG antibodies and neuropathy is supported because pathologic studies of sural nerve biopsies of patients with neuropathy and anti-MAG IgM monoclonal gammopathy show demyelination associated with IgM deposits on the affected myelin sheaths. We conclude that testing for serum autoantibodies should never be the first step in the work-up of peripheral neuropathy but an addi-tional diagnostic measure after careful clinical and electrophysiological evaluation. High quality standards for a diagnostic antibody test should be applied. In our experience anti-MAG antibodies are a valuable marker that is relevant for diagnosis. Patients with chronic demyelinating polyneuro-pathies should be screened for anti-MAG anti-bodies. We test all our patients with lower motor neuron disease or motor neuropathies for anti-GM1 antibodies. We do not routinely test for anti-GM1 or related gangliosides in Guillain-Barré syndrome as the results have not yet a definite impact on the diagnosis or treatment regimen.
Introduction: Anti-ganglioside antibodies (AGA) have been associated with several peripheral neuropathies, such as Miller–Fisher syndrome, Guillain–Barré syndrome and multifocal motor neuropathy. They have also been studied in patients with systemic lupus erythematosus (SLE), focusing on neuropsychiatric manifestations and peripheral neuropathy, but the results are contradictory. Objective: To study the presence of AGA in a large cohort of patients with SLE and neuropsychiatric manifestations. Patients and methods: Serum from 65 consecutive patients with SLE and neuropsychiatric manifestations, collected from 1985 to 2009, was tested for the presence of AGA antibodies (GM1, GM2, GM3, asialo-GM1 GD1a, GD1b, GD3, GT1b, GQ1b) using a standard enzyme-linked immunosorbent assay ELISA test (INCAT 1999) and thin layer chromatography (TLC). Results: Positive results for asialo-GM1 (IgM) were found in 10 patients, 6 were positive for asialo-GM1 (IgM and IgG), and 4 were positive for other AGA such as GM1, GM2, GM3, GD1b, GT1b, GD3, (mainly IgM). Conclusions: Clinical and statistical studies showed no correlation between AGA and neuropsychiatric manifestations of SLE. Although some patients showed reactivity to AGA, these antibodies are not a useful marker of neuropsychiatric manifestations in SLE patients.
THE AIM of the study was the evaluation of autoantibody reaction against endogenous gangliosides in the course of Lyme borreliosis.
Antibodies against profile of gangliosides composed of GM1, GM2, GM3, GD1a,GD1b,GT1b, GQ1b were evaluated in serum patients with early disseminated (neuroborreliosis) Lyme disease (n = 16), patients with long lasting serologic response against Borrelia burgdorferi (n = 32) and in healthy subjects (n = 16). Immunoblot test for IgG was used.
Antibodies were detected in all evaluated groups. In group of neuroborreliosis (lymphocytic meningitis with cranial nerve invoIvement) there was no essential difference with control group. It was stated in group of forestry workers with serological features of infection B. burgdorferi lasting for years.
Results of the study do not support the thesis of participation of IgG autoantibodies against gangliosides in pathogenesis early disseminated Lyme borreliosis in form of lymphocytic meningitis with cranial nerves paresis. Antibodies against endogenous glicosfingolipides in Lyme borreliosis probably can lead to affecting nervous system (demielinisation and polineuropathy) but probably require long-term immunization, what is suggested by results of examined group of patients with the multi-annual serological features of infection.
High titers of serum antibodies to neural antigens occur in several forms of neuropathy. These include neuropathies associated with monoclonal gammopathy, inflammatory polyneuropathies, and paraneoplastic neuropathies. The antibodies frequently react with glycosylated cell surface molecules, including glycolipids, glycoproteins, and glycosaminoglycans, but antibodies to intracellular proteins have also been described. There are several correlations between antibody specificity and clinical symptoms, such as anti-MAG antibodies with demyelinating sensory or sensorimotor neuropathy, anti-GM1 ganglioside antibodies with motor nerve disorders, antibodies to gangliosides containing disialosyl moieties with sensory ataxic neuropathy and Miller–Fisher syndrome, and antibodies to the neuronal nuclear Hu antigens with paraneoplastic sensory neuronopathy. These correlations suggest that the neuropathies may be caused by the antibodies, but evidence for a causal relationship is stronger in some examples than others. In this review, we discuss the origins of the antibodies, evidence for and against their involvement in pathogenic mechanisms, and the implications of these findings for therapy. © 1999 John Wiley & Sons, Inc. Muscle Nerve 22: 800–822, 1999
Anti-ganglioside antibodies are frequently sought in the sera of patients with autoimmune peripheral neuropathy, using an enzyme-linked immunosorbent assay (ELISA) as the principal method for antibody detection. Wide variations in assay performance between laboratores have been reported. In this study, we established a standardized ELISA method between laboratories within the European Inflammatory Neuropathy Cause and Treatment (INCAT) group and determined the inter-laboratory variance in assay performance using both the standardized INCAT method and in-house local methods. As expected, the inter-laboratory variances were greater using local methods than using the standardized method, producing titre estimates which could be 24.8 or 7.6 times larger or smaller, respectively, than the true means for these laboratories. Using the standardized method, the within laboratory measurement error accounted for 41% of the inter-laboratory variation, providing a theoretical upper limit to which technical improvements within laboratories could reduce inter-laboratory variation. These data describe the intrinsic weaknesses within the widely used ganglioside antibody ELISA methods and reinforce the importance of inter-laboratory cooperation within this area. Standardized serological reagents used in this study are available from INCAT members.
Monoclonal IgM kappa from a patient with polyneuropathy associated with paraproteinemia was found to bind to several polysialogangliosides. Binding of IgM to gangliosides was shown by enzyme-linked immunosorbent assay (ELISA) and by overlaying thin-layer chromatograms of human brain and peripheral nerve gangliosides with the patient's serum followed by radioiodinated goat antihuman IgM. The latter technique showed that the IgM paraprotein reacted with a number of gangliosides. In an ELISA the IgM paraprotein reacted strongly with GD2, GD3, GD1b, and GT1b, but not with GM1, GM3, and GD1a. Thus, the epitope for the patient's IgM paraprotein appears to involve the disialosyl configurations.
Serum IgG autoantibodies to GQ1b ganglioside are associated with the acute phase of the Miller-Fisher syndrome (MFS). We investigated the effects of three anti-GQ1b-positive MFS sera in the mouse phrenic-nerve/diaphragm preparation. Miniature endplate potential frequencies increased eight-fold within 25 min, declined rapidly, and ceased altogether after 3 h, when nerve stimulation no longer evoked a response. One MFS convalescent serum (anti-GQ1b negative) and sera from healthy controls and from patients with other neurological diseases were without effect. Thus muscle weakness in MFS may be caused by a serum factor, likely to be GQ1b antibody, that leads to failure of acetylcholine release from motor nerve terminals.
Circulating IgG antibodies to carbohydrate determinants on GQ1b ganglioside are found in the acute phase sera of patients with Miller Fisher syndrome, a variant of Guillain-Barré syndrome. Here we report that the IgG subclass distribution of the anti-GQ1b antibodies is mainly restricted to IgG1 and IgG3 antibodies, subclasses typically associated with a T cell-dependent immune response to protein antigens. This is highly unusual in that IgG responses to carbohydrate determinants are typically of the IgG2 subclass. Anti-GQ1b antibodies also have a limited ability to bind GQ1b in a membrane-like environment, particularly at body temperature. These data suggest that the antigen initiating the immune response in MFS is not likely GQ1b but an unidentified cross-reactive glycoprotein antigen(s). Similar results were obtained for anti-GM1 IgG antibodies in Guillain-Barré syndrome.
Sera from patients with Guillain-Barré syndrome (GBS) frequently have antibodies to various gangliosides. We report a girl with GBS after Mycoplasma pneumoniae infection who had serum IgG antibody to GM1b ganglioside as well as the cold agglutinins. The cold agglutinins are polyclonal IgM autoantibodies to 'I' antigen on erythrocytes. Ganglioside GM1b contains the terminal moiety shared with sialylated I antigen, a main receptor for M. pneumoniae. In this patient, the anti-GM1b antibody may be elicited in a similar mechanism producing anti-I antibody, and functioned in the development of GBS.
The correlation between the activities of anti-GQ1b IgG antibodies in sera of patients with Miller Fisher syndrome and their clinical manifestations was investigated. The clinical severity was analyzed for 16 patients by scoring their symptoms during the peak of the clinical course and 4 weeks after onset. Serum samples were obtained from all the patients within the first 2 weeks of onset and from 12 of them 4 weeks after onset. Samples were immunostained by TLC after which antibody activities were measured by the ELISA. Anti-GQ1b IgG antibody activities reflect the severity of the patients' symptoms, especially ophthalmoplegia. Lessening of symptoms, expressed by the difference in MFS severity scores of the two tests, occurred with the marked decrease in antibodies. Results of cerebrospinal fluid and nerve conduction studies, performed during the first 2 weeks, showed no fluctuation with changes in the clinical symptoms or antibody activities.