RESUMEN
Assessment of the potential allergenicity of novel proteins, including those expressed in genetically modified plants, is an important issue. In previous studies, we have shown that the IgE measurement induced by systemic exposure of BALB/c mice to a range of proteins correlates broadly with what is known of their allergenic potential in humans. The approach used a homologous passive cutaneous anaphylaxis (PCA) assay that reflects IgE-dependent biological activity and is of sufficient sensitivity to detect IgE production in the absence of adjuvant. In previous studies, the immunization phase was conducted independently in two separate facilities, and the subsequent analytical work (PCA) conducted in a single facility. The purpose here was to further evaluate the transferability of this approach. To this end, BALB/c mice were exposed to a range of doses of peanut agglutinin or ovalbumin, allergenic proteins of peanut and hen's egg, respectively, in two independent laboratories. Serial doubling dilutions of serum pooled for each treatment group were analyzed for specific IgE. At higher doses of allergen very similar, or identical, IgE titers were achieved in both laboratories, although at lower doses, responses were somewhat more variable. These data demonstrate that, although technically demanding, the measurement of protein allergen-induced IgE antibody production in mice using PCA is relatively robust and is transferable and reproducible between laboratories. This approach may provide a useful tool for the safety assessment of novel proteins and suggests that continued evaluation of the approach with a wider range of protein allergens and non-sensitising proteins is justified.
Asunto(s)
Alérgenos/inmunología , Proteínas en la Dieta/normas , Pruebas Inmunológicas/normas , Laboratorios/normas , Proteínas/inmunología , Animales , Proteínas en la Dieta/inmunología , Relación Dosis-Respuesta a Droga , Femenino , Inmunoglobulina E/análisis , Inmunoglobulina E/biosíntesis , Inmunoglobulina E/inmunología , Inmunoglobulina G/análisis , Inmunoglobulina G/inmunología , Ratones , Ratones Endogámicos BALB C , Ovalbúmina/inmunología , Anafilaxis Cutánea Pasiva , Aglutinina de Mani/inmunología , Medición de RiesgoRESUMEN
The authors present a case of pyogenic granuloma (botriomycome) of the vocal cord secondary to naso-tracheal intubation. Along with a physiological review, the histological and clinical characteristics of vocal cord pyogenic granuloma are discussed. Preventative measures and a treatment protocol are proposed.
Asunto(s)
Granuloma Piogénico/patología , Enfermedades de la Laringe/patología , Pliegues Vocales/patología , Adulto , Femenino , Granuloma Piogénico/prevención & control , Granuloma Piogénico/terapia , Humanos , Enfermedades de la Laringe/prevención & control , Enfermedades de la Laringe/terapia , PronósticoRESUMEN
Sensory neuropathies are a frequent and dose-limiting complication resulting from treatment with cisplatin. Neurotrophin-3 (NT-3) promotes the survival of the large fiber sensory neurones that are impaired in cisplatin-induced neuropathy, and may therefore serve as a preventive agent. However, the short half-life of recombinant NT-3 after systemic administration limits its clinical applications. We compared two muscle-based gene transfer strategies for the continuous delivery of NT-3 to the bloodstream in an experimental model of cisplatin-induced neuropathy. Electrophysiological studies showed that the intramuscular injection of an adenovirus encoding NT-3 partially prevented the cisplatin-induced increase in sensory distal latencies. Similar effects were observed in cisplatin-treated mice that received intramuscular injections of a plasmid encoding NT-3 associated with in vivo electroporation. The two techniques were well tolerated and induced only slight muscle toxicity. Measurement of renal function, weight and survival showed that neither technique increased the toxicity of cisplatin. Our study shows that gene therapy, using either a viral or a non-viral vector, is a promising strategy for the prevention of cisplatin-induced neuropathy.
Asunto(s)
Antineoplásicos/efectos adversos , Cisplatino/efectos adversos , Técnicas de Transferencia de Gen , Terapia Genética/métodos , Trastornos de la Sensación/prevención & control , Adenoviridae/genética , Animales , Electroporación , Vectores Genéticos , Ratones , Neurotrofina 3/genética , Neurotrofina 3/metabolismo , Trastornos de la Sensación/inducido químicamenteRESUMEN
The electrotransfer of naked DNA has recently been adapted to the transduction of skeletal muscle fibers. We investigated the short- and long-term efficacy of this methodology in wild-type animals and in mouse models of congenital muscular dystrophy (dy/dy, dy(2J)/dy(2J)), or Duchenne muscular dystrophy (mdx/mdx). Using a reporter construct, the short-term efficacy of fiber transduction reached 40% and was similar in wild-type, dy/dy and dy(2J)/dy(2J) animals, indicating that ongoing muscle fibrosis was not a major obstacle to the electrotransfer-mediated gene transfer. Although the complete rejection of transduced fibers was observed within 3 weeks in the absence of immunosuppression, the persistency was prolonged over 10 weeks when transient or continuous immunosuppressive regimens were used. Using therapeutic plasmids, we demonstrated that electrotransfer also allowed the transduction of large constructs encoding the laminin alpha2 chain in dy/dy mouse, or a chimeric dystrophin-EGFP protein in mdx/mdx mouse. The correct sarcolemmal localization of these structural proteins demonstrated the functional relevance of their expression in vivo, with a diffusion domain estimated to be 300 to 500 microm. However, degeneration-regeneration events hampered the long-term stability of transduced fibers. Given its efficacy for naked DNA transfer in these models of muscular dystrophies, and despite some limitations, gene electrotransfer methodology should be further explored as a potential avenue for treatment of muscular dystrophies.
Asunto(s)
Distrofina/genética , Electroporación/métodos , Terapia Genética/métodos , Laminina/genética , Distrofias Musculares/terapia , Animales , Expresión Génica , Proteínas Fluorescentes Verdes , Miembro Posterior , Proteínas Luminiscentes/genética , Ratones , Ratones Endogámicos mdx , Modelos Animales , Músculo Esquelético/metabolismo , Distrofias Musculares/congénito , Distrofias Musculares/metabolismo , Distrofia Muscular de Duchenne/metabolismo , Distrofia Muscular de Duchenne/terapia , Plásmidos , Factores de TiempoRESUMEN
Cisplatin-induced sensory peripheral neuropathy is the dose-limiting factor for cisplatin chemotherapy. We describe the preventive effect of NT-3 delivery, using direct gene transfer into muscle by in vivo electroporation in a mouse model of cisplatin-induced neuropathy. Cisplatin-induced neuropathy was produced by weekly injections of cisplatin (five injections). Two doses of plasmid DNA encoding murine NT-3 (pCMVNT-3) were tested (5 and 50 microg/animal/injection). Cisplatin-treated mice were given two intramuscular injections. The first injection of pCMVNT-3 was given 2 days before the first injection of cisplatin and the second injection 2 weeks later. Six weeks after the start of the experiment, measurement of NT-3 levels (ELISA) demonstrated significant levels both in muscle and plasma. We observed a smaller cisplatin-related increase in the latency of the sensory nerve action potential of the caudal nerve in pCMVNT-3-treated mice than in controls (p < 0.0001). Mean sensory distal latencies were not different between the 5- and 50- microg/animal/injection groups. Treatment with gene therapy induced only a slight muscle toxicity and no general side effects. Therefore, neurotrophic factor delivery by direct gene transfer into muscle by electroporation is of potential benefit in the prevention of cisplatin-induced neuropathy and of peripheral neuropathies in general.
Asunto(s)
Antineoplásicos/efectos adversos , Cisplatino/efectos adversos , Electroporación/métodos , Técnicas de Transferencia de Gen , Músculos/metabolismo , Neuronas Aferentes/patología , Neurotrofina 3/genética , Enfermedades del Sistema Nervioso Periférico/prevención & control , Animales , Nitrógeno de la Urea Sanguínea , Ensayo de Inmunoadsorción Enzimática , Expresión Génica , Técnicas para Inmunoenzimas , Inyecciones Intramusculares , Ratones , Neurotrofina 3/metabolismo , Enfermedades del Sistema Nervioso Periférico/inducido químicamente , Enfermedades del Sistema Nervioso Periférico/metabolismo , PlásmidosRESUMEN
Neurotrophic factors (NFs) are promising agents for the treatment of peripheral neuropathies such as diabetic neuropathy. However, the value of treatment with recombinant NF is limited by the short half-lives of these molecules, which reduces efficiency, and by their potential toxicity. We explored the use of intramuscular injection of a recombinant adenovirus encoding NT-3 (AdNT-3) to deliver sustained low doses of NT-3. We assessed its effect in two rat models: streptozotocin (STZ)-induced diabetes, a model of early diabetic neuropathy characterized by demyelination, and acrylamide experimental neuropathy, a model of diffuse axonal neuropathy which, like late-onset human diabetic neuropathy, results in a diffuse sensorimotor neuropathy with dysautonomy. Treatment of STZ-diabetic rats with AdNT-3 partially prevented the slowing of motor and sensory nerve conduction velocities (p < 0.01 and p < 0.0001, respectively). Treatment with AdNT-3 of acrylamide-intoxicated rats prevented the slowing of motor and nerve conduction velocities (p < 0.001 and p < 0.0001, respectively) and the decrease in amplitude of compound muscle potentials (p < 0.0001), an index of denervation. Acrylamide-intoxicated rats treated with NT-3 had higher than control levels of muscle choline acetyltransferase activity (p < 0.05), suggesting greater muscle innervation. In addition, treatment of acrylamide-intoxicated rats with AdNT-3 significantly improved behavioral test results. Treatment with AdNT-3 was well tolerated with minimal muscle inflammation and no detectable general side effects. Therefore, our results suggest that NT-3 delivery by adenovirus-based gene therapy is a promising strategy for the prevention of both early diabetic neuropathy and axonal neuropathies, especially late axonal diabetic neuropathy.
Asunto(s)
Neuropatías Diabéticas/prevención & control , Fármacos Neuroprotectores , Neurotrofina 3/genética , Acrilamidas/efectos adversos , Animales , Colina O-Acetiltransferasa/metabolismo , Diabetes Mellitus Experimental , Electrofisiología , Expresión Génica , Técnicas de Transferencia de Gen , Terapia Genética , Humanos , Inyecciones Intramusculares , Masculino , Ratas , Ratas Sprague-Dawley , Estreptozocina/administración & dosificación , TransgenesRESUMEN
The neuroprotective drug riluzole (Rilutek) is a sodium channel blocker and anti-excitotoxic drug which is marketed for the treatment of amyotrophic lateral sclerosis (ALS). Previous studies have shown that riluzole prolongs survival of transgenic mice harboring the mutated form of Cu,Zn-superoxide dismutase found in familial forms of the human disease. In this study we have examined the effect of treatment with riluzole in mice suffering from progressive motor neuronopathy (pmn), a hereditary autosomal recessive wasting disease which shares some symptoms of ALS. These mutants display hind limb weakness starting during the 3rd week of life and leading to paralysis and death during the 7th week of life. Daily treatment with 8 mg/kg of riluzole by oral route significantly retarded the appearance of paralysis, increased life span and improved motor performance on grip test and electromyographic results in the early stage of the disease. There was no effect of riluzole on weight gain. These data demonstrate that riluzole significantly prolongs life span, retards the onset of paralysis and slows the evolution of functional parameters connected with muscle strength in the pmn mouse model of motor neuron disease.
Asunto(s)
Enfermedad de la Neurona Motora/tratamiento farmacológico , Debilidad Muscular/prevención & control , Riluzol/farmacología , Tasa de Supervivencia , Animales , Peso Corporal/efectos de los fármacos , Modelos Animales de Enfermedad , Fuerza de la Mano/fisiología , Ratones , Ratones Transgénicos/genética , Enfermedad de la Neurona Motora/fisiopatología , Debilidad Muscular/tratamiento farmacológico , Debilidad Muscular/fisiopatología , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/fisiopatología , Resultado del TratamientoRESUMEN
Overexpression of the ErbB2 and epidermal growth factor receptor (EGFR) tyrosine kinases is frequently observed in squamous cell carcinomas of the head and neck, and has been correlated with shorter overall survival. By immunoblot analysis, we have found EGFR and ErbB2 expression in 6 out of 6 established head and neck cancer cell lines. Elevated EGFR protein levels were noted in 3 and elevated ErbB2 levels in 5 of them. Significant expression of EGFR and ErbB2 was also detected in 17 of 47 and 26 of 45 primary tumor samples. Due to their enhanced expression on the tumor cell surface, these receptors can be regarded as suitable targets for directed cancer therapy. We have analyzed the antitumoral activity of recombinant single-chain antibody toxins specific for ErbB2 and EGFR against head and neck cancer cells in vitro and in vivo. The recombinant toxins consist of the variable domains of the heavy and light chains of monoclonal antibodies (MAbs) genetically fused to a truncated Pseudomonas exotoxin A (ETA). At low concentrations, the ErbB2-specific single-chain antibody (scFv) toxin scFv(FRP5)-ETA and the EGFR-specific toxins scFv(225)-ETA and scFv(14E1)-ETA inhibited the in vitro growth of established head and neck cancer cell lines and primary tumor cells. In a nude mouse tumor model, intratumoral injection of the antibody toxins resulted in the rapid regression of subcutaneously growing CAL 27 tumor xenografts, with scFv(FRP5)-ETA and scFv(14E1)-ETA treatment being most effective and leading to the cure of up to 50% of the animals. Our results suggest that EGFR and ErbB2-specific antibody toxins may become valuable therapeutic reagents for the treatment of squamous cell carcinomas of the head and neck.
Asunto(s)
ADP Ribosa Transferasas , Toxinas Bacterianas , Carcinoma de Células Escamosas/patología , Receptores ErbB/inmunología , Exotoxinas/farmacología , Neoplasias de Cabeza y Cuello/patología , Inmunotoxinas/farmacología , Receptor ErbB-2/inmunología , Factores de Virulencia , Animales , Anticuerpos Monoclonales , Carcinoma de Células Escamosas/química , Carcinoma de Células Escamosas/tratamiento farmacológico , División Celular , Receptores ErbB/análisis , Exotoxinas/administración & dosificación , Exotoxinas/uso terapéutico , Neoplasias de Cabeza y Cuello/química , Neoplasias de Cabeza y Cuello/tratamiento farmacológico , Humanos , Inmunotoxinas/inmunología , Inmunotoxinas/uso terapéutico , Ratones , Ratones Desnudos , Trasplante de Neoplasias , Receptor ErbB-2/análisis , Proteínas Recombinantes de Fusión/inmunología , Proteínas Recombinantes de Fusión/farmacología , Células Tumorales Cultivadas , Exotoxina A de Pseudomonas aeruginosaRESUMEN
Animal models of human disease are important in unravelling the pathophysiology of the condition, for exploring the natural history of disease and for evaluating potential therapies. The development of animal models of human neurodegenerative disease such as ALS is particularly challenging, given the paucity of knowledge of their aetiology and the organizational specificity of the human motor system. Nonetheless, a range of spontaneously occurring, experimentally produced, or genetically engineered models of ALS are now available. Although not always a perfect replica of the ALS disease, these models are shown to be of outstanding importance for investigations of the mechanisms of dysfunction/death of motor neurons in vivo. This is particularly true for the transgenic mouse models expressing superoxide dismutase or cytoskeletal proteins. This approach has provided an unparalleled opportunity for testing of potential pharmacological or gene therapies, and it can be expected that the results of these studies will be translated into the clinical advances of the next years.
Asunto(s)
Enfermedad de la Neurona Motora/fisiopatología , Animales , Modelos Animales de Enfermedad , Humanos , Ratones , Ratones Mutantes Neurológicos , Ratones Transgénicos , Enfermedad de la Neurona Motora/terapia , Neuronas Motoras/efectos de los fármacos , Neuronas Motoras/patología , Neurotoxinas/toxicidad , Superóxido Dismutasa/genéticaRESUMEN
We show the ability of the BSA-trehalose film to convert normally fragile proteins such as mouse monoclonal antibody to the Alzheimer precursor protein A4 (APP695) and cell line TE671 acetylcholine receptor (AChRTE671) into a stable reagent, after its immobilization on microtitration plates. The remarkable property of the dry immobilized proteins are their stability under prolonged exposure to temperatures as high as 50 degrees C. Using the AChRTE671, the proposed method was applied for the measurement of anti-AChR autoantibodies in Myasthenia gravis by means of an enzyme-linked immunosorbent assay (ELISA). The test was shown to be specific and able to detect anti-AChR autoantibodies at concentrations as low as 3 nM. Using the same AchRTE671 as antigen, the results of examination of 34 serum samples for detection of anti-AChR autoantibodies by ELISA were compared with those of the conventional radioimmunoprecipitation assay (RIA). It was concluded that ELISA is another useful method for the diagnosis of M. gravis. The ELISA method offers a rapid, simple, safe and inexpensive means for mass screening of M. gravis.
Asunto(s)
Autoanticuerpos/análisis , Ensayo de Inmunoadsorción Enzimática/métodos , Miastenia Gravis/inmunología , Línea Celular , Ensayo de Inmunoadsorción Enzimática/instrumentación , Humanos , Sensibilidad y EspecificidadRESUMEN
Motor neuron diseases such as amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy cause progressive paralysis, often leading to premature death. Neurotrophic factors have been suggested as therapeutic agents for motor neuron diseases, but their clinical use as injected recombinant protein was limited by toxicity and/or poor bioavailability. We demonstrate here that adenovirus-mediated gene transfer of neurotrophin-3 (NT-3) can produce substantial therapeutic effects in the mouse mutant pmn (progressive motor neuronopathy). After intramuscular injection of the NT-3 adenoviral vector, pmn mice showed a 50% increase in life span, reduced loss of motor axons and improved neuromuscular function as assessed by electromyography. These results were further improved by coinjecting an adenoviral vector coding for ciliary neurotrophic factor. Therefore, adenovirus-mediated gene transfer of neurotrophic factors offers new prospects for the treatment of motor neuron diseases.
Asunto(s)
Terapia Genética/métodos , Enfermedad de la Neurona Motora/terapia , Factores de Crecimiento Nervioso/uso terapéutico , Proteínas del Tejido Nervioso/uso terapéutico , Adenoviridae/genética , Animales , Animales Recién Nacidos , Factor Neurotrófico Ciliar , Electromiografía , Vectores Genéticos , Inyecciones Intramusculares , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Ratones Mutantes , Enfermedad de la Neurona Motora/mortalidad , Músculos/inervación , Degeneración Nerviosa/efectos de los fármacos , Factores de Crecimiento Nervioso/genética , Proteínas del Tejido Nervioso/genética , Neurotrofina 3 , Nervio Frénico/patología , Análisis de SupervivenciaRESUMEN
Dominant mutations of human Cu/Zn superoxide dismutase (SOD1) are found in about 20% of patients with familial amyotrophic lateral sclerosis (FALS). A transgenic mouse model of FALS (FALSG93A mice) has been generated by overexpression of a mutated form of SOD1. Using electromyography we first show that FALSG93A mice suffer from motoneurone dysfunction similar to that observed in ALS patients and fulfill Lambert's criteria for ALS. We also showed that FALSG93A mice demonstrate a massive loss of functional motor units starting at 47 days of age. Impairment of motor neurone function preceeds by 6 weeks the onset of apparent clinical signs (shaking, tremor) and the beginning of motor neurone loss. Neuromuscular deficits in FALS mice do not result from motoneuronal cell death but rather from loss of axonal integrity.
Asunto(s)
Envejecimiento/fisiología , Esclerosis Amiotrófica Lateral/fisiopatología , Unión Neuromuscular/fisiología , Neuronas/patología , Envejecimiento/patología , Esclerosis Amiotrófica Lateral/genética , Esclerosis Amiotrófica Lateral/patología , Animales , Recuento de Células , Desnervación , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Electromiografía , Ratones , Ratones Transgénicos , Regeneración Nerviosa/fisiología , Conducción Nerviosa/fisiología , Sinapsis/fisiologíaRESUMEN
The mouse autosomal recessive mutation progressive motor neuronopathy (pmn) results in early onset motor neuron disease with rapidly progressive hindlimb paralysis, severe muscular wasting, and death at around 6 weeks of age. This mutant provides opportunities for testing novel therapeutic strategies, including the administration of trophic factors, to prevent the degeneration of diseased neurons. The construction of a strain expressing the pmn and the Extra-toe (Xt) phenotypes allows the detection, and therefore the treatment, of affected progeny before the onset of the clinical weakness. Electromyography is the most appropriate technique for a longitudinal study in which a given individual is examined repeatedly. We present the results of an electrophysiological and behavioral exploration of the pmn disease and show that electromyography is a powerful tool for following the course of the disease and evaluating potential therapies relevant to motor neuron diseases.
Asunto(s)
Neuropatía Hereditaria Motora y Sensorial/fisiopatología , Potenciales de Acción , Animales , Diafragma/fisiopatología , Modelos Animales de Enfermedad , Electromiografía , Nervio Facial/fisiopatología , Femenino , Deformidades Congénitas del Pie/genética , Genes Recesivos , Ligamiento Genético , Neuropatía Hereditaria Motora y Sensorial/genética , Masculino , Ratones , Ratones Mutantes Neurológicos , Actividad Motora , Neuronas Motoras/fisiología , Músculo Esquelético/fisiopatología , Conducción Nerviosa , Fenotipo , Tiempo de Reacción , Nervio Ciático/fisiopatología , Dedos del Pie/anomalíasRESUMEN
The embryologic origin of the parietal bone is membranous bone. Experimentally, membranous bone grafts, maintain its original size, structure with good osteo-integration contrary to endochondral bone autografted (ilium, rib or tibia). The morbidity of skull bone grafts is much less than other donor site of bone. In our study, between 1988 and 1992, autogenous split calvarial bone grafts were used in maxillo-facial reconstruction of seventy-one patients. We observed no major complication. Patient followup has been from 2 to 6 years and results are better than endochondral grafts (iliac crest, ribs, tibia). Parietal grafts bone have been used in the following case: nasal reconstruction, rehabilitation of defects in the floor of the orbit, maxilla grafts, correction of frontal depression and mandibular reconstruction.
Asunto(s)
Huesos Faciales/cirugía , Hueso Parietal/trasplante , Adolescente , Adulto , Femenino , Estudios de Seguimiento , Hueso Frontal/cirugía , Humanos , Masculino , Persona de Mediana Edad , Rinoplastia/métodos , Trasplante AutólogoRESUMEN
The use of acetylcholine receptors (AChR) from the readily available TE671 cell line as a practical alternative to human muscle for monitoring the anti-AChR antibody assay in sera of Myasthenia gravis patients has been recently proposed. Most of the TE671 culture protocols include the use of glucocorticoids. Glucocorticoids were shown to upregulate the acetylcholine receptor expression in TE671 cells. To confirm the advantage of using AChR from TE671 cells (AChRTE) and to validate the use of AChR from glucocorticoid-treated cells (AChRGT) in AChR antibody measurement, the three different antigens (muscle AChR (AChRMU), AChRTE, and AChRGT) were compared for radioimmunoprecipitation assay. We found that, despite a slight underestimation of the antibody titers using AChRTE and AChRGT compared to AChRMU, and considering the rare cases of AChRMU antibody titer category permutations, the correlations between the values were satisfactory.
Asunto(s)
Antígenos , Autoanticuerpos/análisis , Dexametasona/farmacología , Músculo Esquelético/química , Miastenia Gravis/inmunología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Autoanticuerpos/sangre , Línea Celular , Femenino , Humanos , Masculino , Persona de Mediana Edad , Miastenia Gravis/sangre , Radioinmunoensayo , Receptores Colinérgicos/inmunología , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
The use of osseointegrated titanium implants has improved the quality of auricle prosthesis fixation. Between december 1993 and july 1994, twenty two titanium fixtures have been surgically implanted in nine patients in the Ear Nose Throat department of the university hospital of Strasbourg. This method of auricle amputation repair achieves a good esthetic result with a low rate of postoperative complications.
Asunto(s)
Oído Externo , Oseointegración , Prótesis e Implantes , Adolescente , Adulto , Niño , Oído Externo/anomalías , Oído Externo/lesiones , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , TitanioRESUMEN
Closure of wide palate defects is still a very complex problem and a real challenge for cervico-facial surgeons. The authors present an original technics drifted of the one D.W. Furnas describe in 1986. It allows, in a single stage, reconstruction of palatal defects. Surgical procedure of the temporal fasciosseous island flap is presented and then, advantages and inconveniences are discussed.
Asunto(s)
Neoplasias Maxilares/cirugía , Hueso Paladar/cirugía , Hueso Parietal/cirugía , Colgajos Quirúrgicos , Estudios de Seguimiento , Humanos , Masculino , Maxilar/cirugía , Neoplasias Maxilares/secundario , Persona de Mediana Edad , Osteotomía/métodosRESUMEN
A mouse model of MG, termed experimental autoimmune myasthenia gravis (EAMG), can be obtained after immunization with Torpedo acetylcholine receptor (AChR). Although many studies have detailed the consequence of AChR antibodies binding at the neuromuscular junction and the difficulty in obtaining obvious clinical signs, less attention has been focused on the possibility of amplifying the muscular block in order to discriminate between immunized and healthy animals. In the present studies we observe that a single inoculation of alpha-bungarotoxin (alpha-bgt) can amplify the neuromuscular block revealed by repetitive nerve stimulation, and induce in EAMG mice a stable muscular weakness state lasting for at least 169 hours instead of 95 hours in normal mice. This model could provide an excellent tool for evaluating drugs active on neuromuscular transmission.
Asunto(s)
Enfermedades Autoinmunes/inducido químicamente , Bungarotoxinas/administración & dosificación , Modelos Animales de Enfermedad , Inmunización , Miastenia Gravis/inducido químicamente , Animales , Enfermedades Autoinmunes/fisiopatología , Femenino , Ratones , Ratones Endogámicos C57BL , Miastenia Gravis/fisiopatología , Receptores Colinérgicos , Transmisión Sináptica/fisiologíaRESUMEN
The measurement of antibodies directed against the cholinergic receptor, produced by myasthenic patients, involves a radioimmunoassay. The use of human muscular nicotinic cholinergic receptor is highly recommended for this assay. However, difficulties in supplying and standardizing this reagent led us to consider its replacement by the homologous cholinergic receptor constitutively expressed by the human cell line TE671. TE671 cells were grown in mass cultures on microcarriers and acetylcholine receptors were solubilized using detergents. Replacement of human muscular receptor by cellular receptor in the radioimmunoassay assay was assessed by challenging these 2 antigens for numeric or diagnostic correlations. Using TE671 receptor, the correlations observed were good and, despite a slight decrease in the absolute values of the titers, about 90 p. 100 of the assays provided the same diagnostic accuracy. Some interfering proteins, perhaps immature acetylcholine receptor alpha subunits, could be expressed and could account for the slight decline in the antibody titers obtained using TE671 receptor. These interferences could be overcome by a further step of purification and concentration of the various receptor preparations. Finally, TE671 receptor could definitely replace human muscular receptor for screening and measuring myasthenic patients auto-antibodies.