RESUMEN
Multifocal motor neuropathy is a rare immune-mediated neuropathy characterized by progressive asymmetric weakness and atrophy without sensory abnormalities. Although disease onset is usually in adulthood, a few childhood-onset cases have been reported. Here, we report the case of an 8-year-old boy with multifocal motor neuropathy who presented with a slowly progressive left and distal upper limb weakness without sensory loss. The initial high-dose intravenous immunoglobulin treatment significantly improved left upper limb muscle weakness. Continued monthly intravenous immunoglobulin treatment gradually improved muscle strength for several months initially. While the muscle strength decreased slightly after 8 months of therapy, it was better than that before intravenous immunoglobulin treatment. One year and eight months after the initiation of treatment, serum testing for IgM antibodies to gangliosides, GM1 and GM2, was negative. This is the first pediatric report of the serum IgM autoantibodies positive to GM1 and GM2. The clinical course is similar to that of partial intravenous immunoglobulin responders among patients with adulthood-onset multifocal motor neuropathy. Since the symptoms plateaued after the initial intravenous immunoglobulin therapy, prognosis appears to be determined by the patient's initial response to intravenous immunoglobulin treatment.
Asunto(s)
Gangliósido G(M1)/inmunología , Gangliósido G(M2)/inmunología , Inmunoglobulina M/sangre , Enfermedades Neurodegenerativas/inmunología , Enfermedades Neuromusculares/inmunología , Niño , Diagnóstico Diferencial , Gangliósido G(M1)/sangre , Gangliósido G(M2)/sangre , Humanos , Inmunoglobulinas Intravenosas/uso terapéutico , Factores Inmunológicos/uso terapéutico , Masculino , Enfermedades Neurodegenerativas/sangre , Enfermedades Neurodegenerativas/diagnóstico , Enfermedades Neurodegenerativas/tratamiento farmacológico , Enfermedades Neuromusculares/sangre , Enfermedades Neuromusculares/diagnóstico , Enfermedades Neuromusculares/tratamiento farmacológicoRESUMEN
GM2 gangliosidosis is a group of inherited neurodegenerative disorders resulting primarily from the excessive accumulation of GM2 gangliosides (GM2) in neuronal cells. As biomarkers for categorising patients and monitoring the effectiveness of developing therapies are lacking for this group of disorders, we sought to develop methodology to quantify GM2 levels in more readily attainable patient samples such as plasma, leukocytes, and cultured skin fibroblasts. Following organic extraction, gangliosides were partitioned into the aqueous phase and isolated using C18 solid-phase extraction columns. Relative quantification of three species of GM2 was achieved using LC/ESI-MS/MS with d35GM1 18:1/18:0 as an internal standard. The assay was linear over the biological range, and all GM2 gangliosidosis patients were demarcated from controls by elevated GM2 in cultured skin fibroblast extracts. However, in leukocytes only some molecular species could be used for differentiation and in plasma only one was informative. A reduction in GM2 was easily detected in patient skin fibroblasts after a short treatment with media from normal cells enriched in secreted ß-hexosaminidase. This method may show promise for measuring the effectiveness of experimental therapies for GM2 gangliosidosis by allowing quantification of a reduction in the primary storage burden.
Asunto(s)
Cromatografía Líquida de Alta Presión , Gangliósido G(M2)/análisis , Espectrometría de Masas en Tándem , Línea Celular , Fibroblastos/química , Gangliósido G(M2)/sangre , Gangliósido G(M2)/aislamiento & purificación , Humanos , Leucocitos/química , Extracción en Fase Sólida , Espectrometría de Masa por Ionización de Electrospray , beta-N-Acetilhexosaminidasas/metabolismoRESUMEN
To find a new biomarker of Tay-Sachs disease and Sandhoff disease. The lyso-GM2 ganglioside (lyso-GM2) levels in the brain and plasma in Sandhoff mice were measured by means of high performance liquid chromatography and the effect of a modified hexosaminidase (Hex) B exhibiting Hex A-like activity was examined. Then, the lyso-GM2 concentrations in human plasma samples were determined. The lyso-GM2 levels in the brain and plasma in Sandhoff mice were apparently increased compared with those in wild-type mice, and they decreased on intracerebroventricular administration of the modified Hex B. The lyso-GM2 levels in plasma of patients with Tay-Sachs disease and Sandhoff disease were increased, and the increase in lyso-GM2 was associated with a decrease in Hex A activity. Lyso-GM2 is expected to be a potential biomarker of Tay-Sachs disease and Sandhoff disease.
Asunto(s)
Gangliósido G(M2)/análogos & derivados , Enfermedad de Sandhoff/metabolismo , Enfermedad de Tay-Sachs/metabolismo , Adulto , Animales , Biomarcadores/sangre , Biomarcadores/metabolismo , Encéfalo/metabolismo , Proteína Activadora de G (M2)/deficiencia , Gangliósido G(M2)/sangre , Gangliósido G(M2)/metabolismo , Hexosaminidasas/sangre , Humanos , Lactante , Ratones , Enfermedad de Sandhoff/sangre , Enfermedad de Sandhoff/enzimología , Enfermedad de Tay-Sachs/sangre , Enfermedad de Tay-Sachs/enzimologíaRESUMEN
BACKGROUND: Pancreatic adenocarcinoma cells express gangliosides and sialyl Lewis (sLe) antigens. It is not known whether these carbohydrate antigens can be targeted by immunotherapy. The authors measured the expression of GM(2) and sLe antigens on the surface of pancreatic carcinoma cells and the serum levels of total gangliosides, GM(2), and antiganglioside antibodies in patients with pancreatic carcinoma. METHODS: Cell surface GM(2) and sLe antigens were measured by cell suspension enzyme linked immunoadsorbent assay (ELISA) in four pancreatic carcinoma cell lines. Sera from 20 pancreatic carcinoma patients and 20 age- and gender-matched healthy volunteers were analyzed for antiganglioside and anti-sLe immunoglobulin (Ig) M titers by ELISA. Serum levels of total gangliosides and GM(2) also were measured. RESULTS: All cell lines expressed GM(2) and sLe antigens. When compared with age- and gender-matched volunteers, patients had significantly higher serum levels of total gangliosides (25.6 +/- 9.0 mg/dL vs. 15.6 +/- 2.7 mg/dL; P < 0.001), GM(2) (0.278 +/- 0.415 mg/dL vs. 0.013 +/- 0.018 mg/dL; P = 0.02), ELISA units of anti-GM(2) IgM antibody (368 +/- 95 vs. 155 +/- 25; P = 0.04) and anti-GD(1b) IgM antibody (351 +/- 91 vs. 138 +/- 26; P = 0.03), but not anti-sLe(x) IgM (1389 +/- 345 vs. 1081 +/- 224; P = 0.46) or anti-sLe(a) IgM antibody (1097 +/- 253 vs. 1200 +/- 315; P = 0.80). Patients with unresectable tumors had higher serum levels of total gangliosides compared with patients with resectable tumors, and a serum level > 25 mg/dL was found to correlate significantly with poor overall survival (P < 0.02). CONCLUSIONS: Increased serum levels of total gangliosides and GM(2) may reflect shedding or release of gangliosides from the surface of tumor cells. Production of IgM antibody against GM(2) and GD(1b) indicates that these gangliosides are immunogenic antigens that may be potential targets for effective active immunotherapy.
Asunto(s)
Adenocarcinoma/inmunología , Gangliósido G(M2)/sangre , Gangliósidos/sangre , Neoplasias Pancreáticas/inmunología , Adenocarcinoma/fisiopatología , Adenocarcinoma/terapia , Adulto , Anciano , Anciano de 80 o más Años , Ensayo de Inmunoadsorción Enzimática , Femenino , Gangliósido G(M2)/inmunología , Gangliósidos/inmunología , Humanos , Inmunoglobulina M/análisis , Inmunoterapia , Masculino , Persona de Mediana Edad , Neoplasias Pancreáticas/fisiopatología , Neoplasias Pancreáticas/terapia , Células Tumorales CultivadasRESUMEN
We retrospectively evaluated measurement data and clinical relevance of autoantibodies to gangliosides in peripheral neuropathies (PN). The IgG and IgM antiganglioside autoantibodies were determined by our own immunodot-blot assay on membrane and by enzyme-linked immunosorbent assay (Elisa) in sera of 1,342 patients with peripheral neuropathies. Anti-GM1 and anti-GD1b autoantibodies formed a part of the normal autoantibody repertoire and were common place in 12% of normal subjects and in 14% of disease control groups. Polyclonal IgM antiganglioside autoantibodies were detected in chronic PN, polyclonal IgG antiganglioside autoantibodies were detected in acute PN. Polyclonal IgM anti-GM1 and anti-GD1b autoantibodies were detected in 35 patients out of 48 with treatable multifocal motor neuropathy with persistent conduction blocks. These autoantibodies well discriminated between suspected motor peripheral neuropathies and motor neuron diseases (sensitivity 73%, specificity 83%, positive predictive value 60%, negative predictive value 91%). Monoclonal IgM autoantibodies reacted strongly with gangliosides in 15 patients out of 77 with M-IgM neuropathy (19%). M-IgM autoantibodies differed in their fine specificities with different principal target antigens as demonstrated with cross-reactivity. Such findings provide further evidence for a relationship between neurological syndromes and antiganglioside antibody profiles and also suggest that different gangliosides could be principal target antigens such as GM1, GD1b, GT1b, GD1a or GM2. Polyclonal IgG anti-GM1 and anti-GD1b autoantibodies were detected in 21 patients out of 22 with acute motor axonal Guillain-Barré syndrome with antecedent of infection by Campylobacter jejuni, polyclonal IgG anti-GQ1b autoantibodies in 9 patients out of 10 with Miller-Fisher syndrome. Detection of antiganglioside autoantibodies by immunodot-blot assay which is simple and quick in testing a large panel of gangliosides has become very important in the diagnosis and in the choose of expensive therapeutic strategies in chronic or acute autoimmune neuropathies.
Asunto(s)
Autoanticuerpos/sangre , Enfermedades Autoinmunes/diagnóstico , Gangliósidos/inmunología , Enfermedades del Sistema Nervioso Periférico/inmunología , Enfermedad Aguda , Infecciones por Campylobacter/inmunología , Campylobacter jejuni , Enfermedad Crónica , Reacciones Cruzadas , Ensayo de Inmunoadsorción Enzimática , Gangliósido G(M1)/sangre , Gangliósido G(M1)/inmunología , Gangliósido G(M2)/sangre , Gangliósido G(M2)/inmunología , Gangliósidos/sangre , Síndrome de Guillain-Barré/inmunología , Humanos , Immunoblotting , Inmunoglobulina G/análisis , Inmunoglobulina M/análisis , Síndrome de Miller Fisher/inmunología , Enfermedad de la Neurona Motora/inmunología , Factores de Crecimiento Nervioso/inmunología , Conducción Nerviosa/inmunología , Enfermedades del Sistema Nervioso Periférico/diagnóstico , Valor Predictivo de las Pruebas , Estudios Retrospectivos , Sensibilidad y EspecificidadRESUMEN
Guillain-Barré syndrome (GBS) sometimes is preceded by cytomegalovirus (CMV) infection. Irie et al. (J. Neuroimmunol. 1996;68:19-26) reported that three patients with GBS subsequent to CMV infection had IgM and IgG anti-GM2 antibodies. In our larger study, the IgMs from the CMV-associated GBS patients showed significantly higher anti-GM2 antibody titers than anti-GM3, anti-GD1a, anti-GD1b, anti-GD2, anti-GD3, anti-GT1b, anti-GQ1b, and anti-SGPG antibody titers. None of the anti-glycosphingolipid antibody titers differed significantly from the others in the IgGs from the CMV-associated GBS patients. However, IgM anti-GM2 antibody frequently was present in GBS patients who were not preceded by CMV infection and non-GBS patients with acute CMV infection. Our results did not support the conclusion of Irie et al. that anti-GM2 antibodies were closely associated with acute CMV infection in GBS, but acute CMV infection, with and without GBS, was associated with IgM anti-GM2 antibody.
Asunto(s)
Infecciones por Citomegalovirus/inmunología , Gangliósido G(M2)/inmunología , Inmunoglobulina M/inmunología , Polirradiculoneuropatía/inmunología , Enfermedad Aguda , Adolescente , Adulto , Anticuerpos Antivirales/sangre , Anticuerpos Antivirales/inmunología , Cromatografía en Capa Delgada , Citomegalovirus/inmunología , Infecciones por Citomegalovirus/sangre , Infecciones por Citomegalovirus/complicaciones , Ensayo de Inmunoadsorción Enzimática , Femenino , Gangliósido G(M2)/sangre , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Inmunoglobulina M/sangre , Masculino , Persona de Mediana Edad , Polirradiculoneuropatía/sangre , Polirradiculoneuropatía/virologíaRESUMEN
The content of serum gangliosides was examined in VM and C57BL/6J (B6) mice that contained subcutaneous metastatic (VM) and non-metastatic (CT-2A) brain tumors, respectively. Gas-liquid chromatography (GLC) and high performance thin-layer chromatography (HPTLC) were used to analyze the serum gangliosides. N-glycolylneuraminic acid (NeuGc) accounted for greater than 90% of the total serum sialic acid content in each mouse strain (5.53 nmol and 2.05 nmol per ml serum, respectively). GM2-NeuGc was the major serum ganglioside detectable in both the normal and tumor-bearing mice of each strain. Shedding of tumor gangliosides into the serum occurs in various murine non-neural tumors and in human gliomas and neuroblastomas, but has not been previously studied in murine brain tumors. Our results show that serum ganglioside concentration was reduced in VM mice bearing the metastatic VM tumor, but was increased in B6 mice bearing the non-metastatic CT-2A tumor. These changes in concentration, however, were not associated with marked changes in serum ganglioside distribution. As serum gangliosides are synthesized in the liver, the differences in serum ganglioside concentration in the tumor-bearing mice may arise more from changes in liver function than from differences in tumor shedding.
Asunto(s)
Neoplasias Encefálicas/sangre , Neoplasias Encefálicas/secundario , Gangliósidos/sangre , Animales , Neoplasias Encefálicas/metabolismo , Femenino , Gangliósido G(M2)/análogos & derivados , Gangliósido G(M2)/sangre , Gangliósido G(M2)/metabolismo , Gangliósidos/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Trasplante de NeoplasiasRESUMEN
The polymorphic expression of GM4(NeuAc), GM3(NeuGc), GM2(NeuGc), and GM1(NeuGc) was found in erythrocytes of inbred strains of mice [Nakamura, K. et al. (1988) J. Biochem. 103, 201-208]. In this paper, we report the results of genetic analysis of the expression of GM4(NeuAc) and GM2(NeuGc). Ganglioside analysis of the progeny obtained on mating between BALB/c mice [GM4 (+)] and WHT/Ht or C57BL/6 mice [both GM4 (-)] indicated that the expression of GM4(NeuAc) is an autosomal dominant trait, and that WHT/Ht and C57BL/6 mice carry a defect on a single autosomal gene. We named this gene Gsl-4. On quantitative determination of galactosylceramide (GalCer), which is the biosynthetic precursor of GM4(NeuAc), the content of GalCer was found to be quite low in WHT/Ht erythrocytes, compared with in BALB/c erythrocytes. On analysis of GM4(NeuAc) and GalCer in 92 backcross mice produced on mating between BALB/c and WHT/Ht mice, it was found that 45 GM4(+) mice apparently expressed a detectable amount of GalCer and that 47 GM4(-) mice expressed an almost undetectable amount of GalCer. These results suggest that Gsl-4 controls the expression of GM4(NeuAc) by regulating the content of GalCer. Linkage analysis of Gsl-4 and the gene controlling GM2(NeuGc) in erythrocytes indicated that the two genes are not genetically linked. Comparison of the ganglioside expression in liver and erythrocytes of the same backcross mice suggested that the gene controlling GM2(NeuGc) expression in the liver (Ggm-2) is also responsible for the expression of GM2(NeuGc) in erythrocytes.
Asunto(s)
Eritrocitos/metabolismo , Gangliósido G(M2)/genética , Gangliósidos/genética , Regulación de la Expresión Génica , Animales , Cromatografía en Capa Delgada , Mapeo Cromosómico , Gangliósido G(M2)/análogos & derivados , Gangliósido G(M2)/sangre , Galactosilceramidas/sangre , Galactosilceramidas/genética , Gangliósidos/sangre , Ligamiento Genético , Hígado/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BLRESUMEN
The lytic effect of Clostridium perfringens delta toxin was investigated on goat, human, rabbit, and guinea pig platelets. In contrast to erythrocytes from the latter three species, which are insensitive to the toxin, the platelets were equally lysed by the same amount of toxin. These results suggest the presence of GM2 or GM2-like ganglioside(s) as a specific recognition site of the toxin on platelet plasmic membrane as previously established for sensitive erythrocytes. Plasmic membrane damage of human platelets was evidenced by the release of entrapped alpha-[14C]aminoisobutyric acid used as a cytoplasmic marker. The specific binding of hemolytically active 125I-delta toxin by human and rabbit platelets was practically identical, dose dependent, and inhibitable by GM2. Labeled toxin was also bound by various subcellular organelles separated from rabbit platelets except the 5-hydroxytryptamine (5-HT)-containing dense bodies, suggesting the absence or inaccessibility of GM2 on the surface of the latter organelles. This result correlates with the low amounts of 5-[3H]HT liberated after platelet challenge with delta toxin whereas this mediator was massively liberated upon lysis by the sulfhydryl-activated toxin alveolysin. The levels of M and P forms of phenol sulfotransferase (PST), involved in 5-HT catabolism, were determined in human platelet lysates after challenge with delta toxin, alveolysin, and other disruptive treatments. The low PST-M activities detected after lysis by delta toxin suggest that this isoenzyme is very likely associated to dense bodies in contrast to PST-P which is cytoplasmic. Platelet lysis by the toxin allows easy separation of these organelles.
Asunto(s)
Toxinas Bacterianas/farmacología , Plaquetas/efectos de los fármacos , Gangliósido G(M2)/sangre , Gangliósidos/sangre , Ácidos Aminoisobutíricos/sangre , Animales , Arilsulfotransferasa , Proteínas Bacterianas , Cabras , Cobayas , Proteínas Hemolisinas/farmacología , Humanos , Compuestos Orgánicos , Serotonina/sangre , Estreptolisinas/farmacología , Sulfurtransferasas/sangreRESUMEN
GM2 ganglioside was detected in all five sera of hepatoma patients analyzed by thin-layer chromatography in conjunction with enzyme-immunostaining with rabbit anti-GM2 antibody and horseradish peroxidase-conjugated anti-rabbit IgG, and was quantitated by densitometry. The GM2 contents of these sera were 20 to 100 times higher than those of sera from normal adults. By the method used, an increased GM2 level could be detected with samples of less than 0.1 ml of sera from hepatoma patients. The elevated levels of serum GM2 in hepatoma patients were suggested to be due to elevated GM2 levels in the liver lesions, because the GM2 contents of the liver of five patients with liver cancer, including three with hepatoma, were higher than those of normal liver tissues.
Asunto(s)
Carcinoma Hepatocelular/sangre , Gangliósido G(M2)/sangre , Gangliósidos/sangre , Neoplasias Hepáticas/sangre , Cromatografía en Capa Delgada , Técnicas para InmunoenzimasRESUMEN
Concentration of gangliotriaose-series glycosphingolipids, including GA2, GM2, GD2 and GT2, was measured in human sera by a thin-layer chromatography/enzyme-immunostaining method. By this method, as little as 5-10 ng/ml of these glycolipids in serum could be determined simultaneously. Although GD2 ganglioside could be consistently detected in normal cord blood (1-2 ng/ml of serum), the ganglioside was never detected in normal adult serum. However, the same ganglioside was found to be present in large quantity in preoperative sera of 6/9 patients with neuroblastomas (25-658 ng/ml of serum). In addition to GD2, gangliosides GM2 and GA2 increased concomitantly than usual. It is concluded that this highly sensitive quantification of the tumor-associated glycolipids circulating in serum of neuroblastoma patients could be useful in their diagnosis.
Asunto(s)
Sangre Fetal/análisis , Gangliósidos/sangre , Neuroblastoma/sangre , Adulto , Cromatografía en Capa Delgada , Gangliósido G(M2)/sangre , Glicoesfingolípidos/sangre , Humanos , Técnicas para InmunoenzimasRESUMEN
Glycolipids of erythrocytes from inbred mice, C3H/He and CDF1, were analyzed. Galactosylceramide, glucosylceramide, GM4 and GM2 were separated and their structures were established. Sialic acid of GM4 was exclusively N-acetylneuraminic acid, while GM2 contained only N-glycolylneuraminic acid. The fatty acid composition of GM4 was very similar to that of galactosylceramide, and a similarity was also found between that of GM2 and glucosylceramide. These results suggest that erythroid cells of mice have two separate biosynthetic pathways of gangliosides. One is the pathway to GM4 (NeuAc) from galactosylceramide, and the other is that for synthesizing GM2 (NeuGc) from glucosylceramide.
Asunto(s)
Eritrocitos/análisis , Gangliósido G(M2)/sangre , Gangliósidos/sangre , Animales , Cromatografía en Capa Delgada , Galactosilceramidas/sangre , Glucosilceramidas/sangre , Ratones , Ratones Endogámicos C3H , Ratones Endogámicos , Especificidad de la EspecieRESUMEN
Enzyme replacement therapy was attempted with two Tay-Sachs-diseased individuals--a 14-month-old child and a 7-week-old infant. Treatment consisted of repeated weekly intrathecal injections of pure hexosaminidase A. Injection of this enzyme resulted in almost complete disappearance of GM2 from the serum, but did not bring about dissolution of the GM2 membranous cytoplasmic bodies in the brain, as detected by electronmicroscopy. Both patients tolerated the treatment without apparent clinical complications, but no clear-cut improvement was noted as a result of prolonged injections of hexosaminidase A. Since this treatment was initiated in both an advanced stage and a very early stage of the disease, we conclude that enzyme replacement treatment by this route is not beneficial for patients with Tay-Sachs disease.