RESUMO
Recently, the disappearance of oligoclonal bands (OCBs) from the cerebrospinal fluid (CSF) of a few natalizumab-treated patients with multiple sclerosis (MS) has been reported. This is interesting since CSF-restricted OCB are believed to persist in MS. We pooled CSF data from 14 MS centers to obtain an adequate sample size for investigating the suspected changes in central nervous system (CNS)-restricted humoral immune activities in the context of natalizumab therapy. In a retrospective chart analysis, CSF parameters of blood-CSF barrier integrity and intrathecal IgG production from 73 natalizumab-treated MS patients requiring a diagnostic puncture for exclusion of progressive multifocal leukoencephalopathy were compared with CSF data obtained earlier in the course of disease before natalizumab therapy. At the time of repeat lumbar puncture, local IgG production (according to Reibergram) was significantly reduced (p < 0.0001) and OCB had disappeared in 16% of the patients. We therefore conclude that natalizumab therapy interferes with intrathecal antibody production at least in a significant number of patients.
Assuntos
Anticorpos Monoclonais Humanizados/uso terapêutico , Formação de Anticorpos/efeitos dos fármacos , Linfócitos B/imunologia , Imunoglobulina G/líquido cefalorraquidiano , Esclerose Múltipla/líquido cefalorraquidiano , Bandas Oligoclonais/líquido cefalorraquidiano , Adolescente , Adulto , Idoso , Linfócitos B/efeitos dos fármacos , Feminino , Humanos , Imunoglobulina G/imunologia , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/tratamento farmacológico , Esclerose Múltipla/imunologia , Natalizumab , Bandas Oligoclonais/efeitos dos fármacos , Bandas Oligoclonais/imunologia , Estudos Retrospectivos , Adulto JovemRESUMO
Autoantibodies to the synaptic protein amphiphysin play a crucial pathogenic role in paraneoplastic stiff-person syndrome. Impairment of GABAergic inhibition is the presumed pathophysiological mechanism by which these autoantibodies become pathogenic. Here we used calcium imaging on rat embryonic motor neurons to investigate whether antibodies to amphiphysin directly hinder GABAergic signaling. We found that the immunoglobulin G fraction from a patient with stiff-person syndrome, containing high titer antibodies to amphiphysin and inducing stiffness in rats upon passive transfer, reduced GABA-induced calcium influx in embryonic motor neurons. Depletion of the anti-amphiphysin fraction from the patient's IgG by selective affinity chromatography abolished this effect, showing its specificity for amphiphysin. Quantification of the surface expression of the Na(+)/K(+)/2Cl(2-) cotransporter revealed a reduction after incubation with anti-amphiphysin IgG, which is concordant with a lower intracellular chloride concentration and thus impairment of GABA mediated calcium influx. Thus, anti-amphiphysin antibodies exert a direct effect on GABA signaling, which is likely to contribute to the pathogenesis of SPS.
Assuntos
Anticorpos/farmacologia , Cálcio/metabolismo , GABAérgicos/farmacologia , Neurônios Motores/efeitos dos fármacos , Neurônios Motores/metabolismo , Proteínas do Tecido Nervoso/imunologia , Ácido gama-Aminobutírico/farmacologia , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Embrião de Mamíferos , Feminino , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Camundongos , Proteínas do Tecido Nervoso/metabolismo , Gravidez , Ratos , Medula Espinal/citologia , Estatísticas não Paramétricas , Simportadores/metabolismo , Cotransportadores de K e Cl-RESUMO
BACKGROUND: Antiganglioside antibodies have been reported to play a part in the pathophysiology of Guillain-Barré syndrome (GBS). AIMS: To investigate the prevalence and correlation of anti-ganglioside antibodies with clinical data in children with GBS in a multicentre clinical trial. METHODS: Immunoglobin (Ig)G and IgM to GM1, GM1b, GD1a, GalNAc-GD1a, GD1b, GT1a, and GQ1b were measured by ELISA in sera obtained before treatment. In addition, serological testing for Campylobacter jejuni was carried out. In parallel, a group of adults with GBS and a control group of children without GBS or other inflammatory diseases were evaluated. RESULTS: Sera from 63 children with GBS, 36 adults with GBS and 41 children without GBS were evaluated. Four of the children with GBS showed positive IgG to GM1, in one case combined with anti-GalNAc-GD1a and in one with anti-GD1b. Two others showed isolated positive IgG to GD1b and GT1a. One showed increased anti-GalNAc-GD1a IgM. In 5 of the 63 children, serological evidence of a recent infection with C jejuni was found, and this correlated significantly with the raised antibodies (p = 0.001). In the control group without GBS, no child showed positive IgG, but one showed anti-GalNAc-GD1a IgM. Compared with the adults with GBS, the frequency of antibodies in children was insignificantly lower. In our study, patients with positive antibodies did not show a more severe GBS course or worse outcome than those who were seronegative, and we could not show an increased incidence of axonal dysfunction. CONCLUSIONS: In some children with GBS, one can detect raised IgG against various gangliosides, similar to that in adults. A recent infection with C jejuni is markedly associated with the presence of these antibodies. However, in contrast with what has been reported in adults, in this study we were unable to show a negative effect of these findings on the clinical course.
Assuntos
Gangliosídeo G(M1)/imunologia , Síndrome de Guillain-Barré/imunologia , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Infecções por Campylobacter/imunologia , Campylobacter jejuni/imunologia , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Feminino , Gangliosídeo G(M1)/análogos & derivados , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Estudos ProspectivosRESUMO
Using a polypeptide oligomer harboring 16 repeats of the neuritogenic epitope (aa 58-73) of myelin P2 protein separated by spacers, enhancement of the immune response to the P2 protein, an important neuritogenic autoantigen in experimental autoimmune neuritis (EAN), was attempted. In contrast to a previous study with PLP-16-mer antigen-specific response of T cells was attenuated at all doses examined to a variable degree. Treatment of Lewis rats with the P2-16-mer up to 2 months before immunization with P2(53-78) (vaccination) or after immunization but before appearance of disease (prevention) had a strong tolerizing effect against the induction of EAN on immunization with P2(53-78). Moreover, rats injected with 200 microg of the P2-16-mer i.v. on day 11 after disease induction, at which time the initial signs of disease had appeared, were almost completely protected against progression of clinical disease, whereas animals treated with the same amount of monomeric control peptide developed severe disease (treatment). Similar results were obtained by i.v. treatment of adoptive-transfer EAN with the P2-16-mer. The lack of clinical signs of disease after 16-mer therapy could be correlated with a reduced proliferative response of P2(53-78)-specific lymph node cells. The frequency of apoptotic T cells in sciatic nerve or in lymph node cells, however, was not increased by the 16-mer treatment, suggesting that induction of anergy or other forms of peripheral tolerance may be responsible for the effect. Thus, the oligomerized P2 peptide antigen was highly effective in all three treatment modalities examined in this specific autoreactive T cell-mediated immune response.
Assuntos
Autoantígenos/imunologia , Epitopos de Linfócito T/imunologia , Proteína P2 de Mielina/imunologia , Neurite Autoimune Experimental/prevenção & controle , Oligopeptídeos/imunologia , Fragmentos de Peptídeos/imunologia , Vacinação , Vacinas Sintéticas/imunologia , Animais , Apoptose , Autoantígenos/uso terapêutico , Divisão Celular , Linhagem Celular , Regulação para Baixo , Epitopos de Linfócito T/uso terapêutico , Imunoterapia/métodos , Linfonodos/imunologia , Proteína P2 de Mielina/uso terapêutico , Neurite Autoimune Experimental/imunologia , Neurite Autoimune Experimental/terapia , Oligopeptídeos/uso terapêutico , Fragmentos de Peptídeos/uso terapêutico , Ratos , Ratos Endogâmicos Lew , Solubilidade , Linfócitos T/citologia , Linfócitos T/imunologia , Vacinação/métodos , Vacinas Sintéticas/uso terapêuticoRESUMO
Treatment of experimental autoimmune disorders of the nervous system with high doses of glucocorticosteroids (GC) or with administration of the specific antigen is effective and associated with marked T cell apoptosis in situ. Here we investigated in adoptive transfer-experimental autoimmune neuritis (AT-EAN) of the Lewis rat whether induction of T cell apoptosis resulting from T cell activation by antigen therapy can be further augmented by glucocorticosteroids (GC). AT-EAN was induced by intravenous injection of P2-specific T cell blasts. At the maximum of disease two pulses of the antigen recombinant human P2 (rhP2) were given within 12 h. Methylprednisolone was administered simultaneously or 2 h after the antigen and animals were killed 6 h after the second antigen injection. Using an in situ tailing technique followed by immunocytochemical analysis, the presence of DNA fragmentation in T lymphocytes was confirmed. The bromodeoxyuridine (BrdU) technique was employed to detect in situ proliferating cells. T cell apoptosis in sciatic nerve was enhanced after monotherapy with either antigen or GC compared to the control group receiving an irrelevant myelin protein, recombinant human P0. In combination therapy, a synergistic effect on T cell apoptosis in sciatic nerve was obtained when methylprednisolone was injected sequentially, 2 h after rhP2 protein. BrdU incorporation in the sciatic nerve as well as in the spleen, a major lymphoid organ, was significantly enhanced in animals treated with antigen followed by GC 2 h later as compared to rats receiving rhP2 only, speaking for T cell proliferation in situ associated with T cells undergoing apoptosis. Our findings underscore that different proapoptotic stimuli may act synergistically, depending on the timing of the second treatment. In this scenario even local T cell proliferation in the inflamed nervous system occurs. These results support the paradigm of antigen presentation in the nervous system.
Assuntos
Corticosteroides/farmacologia , Apoptose/efeitos dos fármacos , Proteína P2 de Mielina/farmacologia , Neurite Autoimune Experimental/patologia , Neurite Autoimune Experimental/fisiopatologia , Linfócitos T/patologia , Linfócitos T/fisiologia , Animais , Divisão Celular/efeitos dos fármacos , Combinação de Medicamentos , Humanos , Cinética , Ratos , Proteínas Recombinantes , Nervo Isquiático/patologia , Nervo Isquiático/fisiopatologia , Baço/patologiaRESUMO
Schwann cell (SC) apoptosis may be a critical factor challenging nerve remyelination and regeneration in experimental autoimmune neuritis (EAN) in the Lewis rat. We therefore analyzed the fate of SC during high-dose antigen therapy of adoptive transfer-(AT-) EAN using rhP2 protein. P2 antigen therapy was associated with an increase of tumor necrosis factor (serum levels 1 h after intravenous (i.v.) injection and an augmentation of T-cell apoptosis. Antigen specific therapy had no clear effect on SC apoptosis. The effects on SC apoptosis were determined by morphological criteria or by in situ tailing (IST) followed by immunocytochemical analysis. Secondly, we neutralized TNF-alpha, released in abundance by antigen treatment but only in small concentrations during natural disease course. We found that the addition of a TNF-alpha neutralizing antiserum resulted in a significant decrease in the rate of SC apoptosis in vivo compared to animals treated with control antigen rhP0 or with rhP2 only.
Assuntos
Apoptose/efeitos dos fármacos , Síndrome de Guillain-Barré/patologia , Neurite Autoimune Experimental/patologia , Nervos Periféricos/patologia , Células de Schwann/patologia , Fator de Necrose Tumoral alfa/metabolismo , Transferência Adotiva/efeitos adversos , Transferência Adotiva/métodos , Animais , Anticorpos/imunologia , Anticorpos/farmacologia , Apoptose/imunologia , Feminino , Síndrome de Guillain-Barré/tratamento farmacológico , Síndrome de Guillain-Barré/imunologia , Imunoterapia/efeitos adversos , Imunoterapia/métodos , Proteína P2 de Mielina/antagonistas & inibidores , Proteína P2 de Mielina/imunologia , Proteína P2 de Mielina/farmacologia , Neurite Autoimune Experimental/tratamento farmacológico , Neurite Autoimune Experimental/imunologia , Nervos Periféricos/efeitos dos fármacos , Nervos Periféricos/fisiopatologia , Ratos , Ratos Endogâmicos Lew , Células de Schwann/efeitos dos fármacos , Células de Schwann/imunologia , Nervo Isquiático/efeitos dos fármacos , Nervo Isquiático/patologia , Nervo Isquiático/fisiopatologia , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Studies on postmortem tissue of patients with primary adult-onset dystonia revealed a significant increase in copper levels and a reduction of copper transporting Menkes protein of the lentiform nuclei. Here we demonstrate that patients with idiopathic adult-onset cervical dystonia (n = 14) have reduced Menkes mRNA copies and lower copper levels in leukocytes compared to controls (n = 17; U test, p < 0.05). Changes were less distinct in patients with blepharospasm. Therefore, disturbances of copper metabolism in focal dystonia may not be restricted to the basal ganglia.
Assuntos
Adenosina Trifosfatases/sangue , Gânglios da Base/metabolismo , Proteínas de Transporte/sangue , Proteínas de Transporte de Cátions , Cobre/sangue , Distúrbios Distônicos/sangue , RNA Mensageiro/sangue , Proteínas Recombinantes de Fusão , Adulto , Idoso , ATPases Transportadoras de Cobre , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da PolimeraseRESUMO
OBJECTIVE: To test the hypothesis that different preceding infections influence the neurophysiologic classification and clinical features of Guillain-Barré syndrome (GBS). METHODS: We tested pretreatment sera, 7 +/- 3 (mean +/- SD) days from onset, from 229 patients with GBS in a multicenter trial of plasma exchange and immunoglobulin, for serological markers of infection, adhesion molecules, and cytokine receptors, and compared these with neurophysiologic and clinical features. RESULTS: Recent infection by Campylobacter jejuni was found in 53 patients (23%), cytomegalovirus in 19 (8%), and Epstein-Barr virus in four (2%). Patients with C. jejuni infection were more likely than others to have neurophysiologic criteria of axonal neuropathy or inexcitable nerves, antiganglioside GM(1) antibodies, pure motor GBS, lower CSF protein, and worse outcome. Patients with cytomegalovirus infection were younger and more likely than others to have raised serum concentrations of molecules important in T lymphocyte activation and migration, soluble intercellular adhesion molecule-1 (sICAM-1), soluble vascular cell adhesion molecule-1 (sVCAM-1), soluble leukocyte selectin, and soluble interleukin-2 receptor (sIL-2R). Concentrations of sICAM-1 and soluble tumor necrosis factor receptor were higher in patients with inexcitable nerves than those with demyelinating neurophysiology. Logistic regression analysis showed death or inability to walk unaided at 48 weeks were associated with diarrhea, inexcitable nerves, severe arm weakness, age over 50, raised sIL-2R concentration and absence of immunoglobulin (Ig) M antiganglioside GM(1) antibodies. CONCLUSIONS: Subtypes of GBS defined by preceding infections were only approximately associated with different patterns of clinical, neurophysiologic, and immunologic features. A single infectious agent caused more than one type of pathology in GBS, implying interaction with additional host factors. Most patients had no identified infection.
Assuntos
Anticorpos/imunologia , Infecções Bacterianas/imunologia , Moléculas de Adesão Celular/imunologia , Gangliosídeo G(M1)/imunologia , Síndrome de Guillain-Barré/imunologia , Receptores de Citocinas/imunologia , Ensaio de Imunoadsorção Enzimática , Síndrome de Guillain-Barré/microbiologia , Herpesvirus Humano 4/isolamento & purificação , Humanos , Prognóstico , Análise de RegressãoRESUMO
High-dose Ag administration induces apoptotic death of autoreactive T cells and is an effective therapy of experimental autoimmune diseases of the nervous system. To explore the role of cytokines in Ag-specific immunotherapy, we analyzed mRNA induction and protein expression for the proinflammatory cytokines TNF-alpha and IFN-gamma, the anti-inflammatory cytokine IL-10, and the cytokine-inducible NO synthase (iNOS) during high-dose Ag therapy of adoptive transfer experimental autoimmune encephalomyelitis (AT-EAE) in the Lewis rat. Using semiquantitative and competitive RT-PCR, we found 5- to 6-fold induction of TNF-alpha mRNA and 3-fold induction of IFN-gamma mRNA in the spinal cord that occurred within 1 h after i.v. injection of Ag and was accompanied by a 2-fold increase of iNOS mRNA. Both IFN-gamma and iNOS mRNA remained elevated for at least 6 h, whereas TNF-alpha mRNA was already down-regulated 6 h after Ag injection. A comparable time course was found for circulating serum levels of TNF-alpha and IFN-gamma. IL-10 mRNA levels did not change significantly following Ag injection. Neutralization of TNF-alpha by anti-TNF-alpha antiserum in vivo led to a significant decrease in the rate of T cell and oligodendrocyte apoptosis induced by high-dose Ag administration, but did not change the beneficial clinical effect of Ag therapy. Our data suggest profound activation of proinflammatory but not of anti-inflammatory cytokine gene expression by high-dose Ag injection. Functionally, TNF-alpha contributes to increased apoptosis of both autoaggressive T cells and oligodendrocytes in the target organ and may thereby play a dual role in this model of Ag-specific therapy of CNS autoimmune diseases.
Assuntos
Autoantígenos/uso terapêutico , Citocinas/biossíntese , Encefalomielite Autoimune Experimental/imunologia , Encefalomielite Autoimune Experimental/terapia , Proteína Básica da Mielina/imunologia , Proteína Básica da Mielina/uso terapêutico , Óxido Nítrico Sintase/biossíntese , Células Th1/metabolismo , Animais , Apoptose/imunologia , Autoantígenos/administração & dosagem , Movimento Celular/imunologia , Citocinas/sangue , Progressão da Doença , Relação Dose-Resposta Imunológica , Esquema de Medicação , Encefalomielite Autoimune Experimental/enzimologia , Encefalomielite Autoimune Experimental/patologia , Indução Enzimática/genética , Indução Enzimática/imunologia , Feminino , Regulação da Expressão Gênica/imunologia , Cobaias , Soros Imunes/administração & dosagem , Injeções Intravenosas , Proteína Básica da Mielina/administração & dosagem , Óxido Nítrico Sintase/genética , Óxido Nítrico Sintase Tipo II , RNA Mensageiro/biossíntese , Ratos , Ratos Endogâmicos Lew , Medula Espinal/enzimologia , Medula Espinal/imunologia , Medula Espinal/patologia , Células Th1/imunologia , Fator de Necrose Tumoral alfa/imunologiaRESUMO
TNF-alpha has been implicated as a potentially detrimental cytokine in autoimmune disorders of the nervous system and has been reported to be elevated in antigen-specific therapy of experimental autoimmune neuritis (EAN) in vivo. To investigate the role of TNF-alpha in EAN in rats that had been subjected to antigen-specific therapy with human P2 protein, animals were cotreated with an anti-TNF-alpha neutralizing antibody and the effects of the antibody on disease determined. Using this strategy in adoptive transfer (AT-) EAN, antigen-induced T-cell apoptosis in inflamed sciatic nerve and in liver was reduced to levels observed in control animals indicating that TNF-alpha mediates antigen-induced apoptosis of inflammatory T-cells. Focal liver necrosis, which had been observed in earlier studies after antigen therapy in AT-EAN, was prevented by passive immunization with neutralizing anti-TNF-alpha antibody. Unexpectedly, neutralization of TNF-alpha only partly abolished the protective effect of antigen therapy on the overall disease course. This may indicate that inhibition of TNF-alpha exerts beneficial effects other than through T-cell apoptosis, or that some of the benefit of antigen therapy is mediated by other pathways. These results indicate that secretion of TNF-alpha during antigen therapy has the dual potential to mediate beneficial apoptosis of inflammatory T-cells in the inflammatory lesion and to induce liver damage as a severe side effect.
Assuntos
Antígenos/administração & dosagem , Neurite Autoimune Experimental/tratamento farmacológico , Neurite Autoimune Experimental/fisiopatologia , Fator de Necrose Tumoral alfa/fisiologia , Animais , Antígenos/efeitos adversos , Antígenos/uso terapêutico , Apoptose/efeitos dos fármacos , Linhagem Celular , Doença Hepática Induzida por Substâncias e Drogas , Relação Dose-Resposta a Droga , Feminino , Humanos , Fígado/patologia , Fígado/fisiopatologia , Hepatopatias/prevenção & controle , Proteína P2 de Mielina/efeitos adversos , Proteína P2 de Mielina/uso terapêutico , Necrose , Ratos , Ratos Endogâmicos Lew , Proteínas Recombinantes/efeitos adversos , Proteínas Recombinantes/uso terapêutico , Nervo Isquiático/patologia , Nervo Isquiático/fisiopatologia , Linfócitos T/metabolismo , Linfócitos T/fisiologia , Fator de Necrose Tumoral alfa/efeitos adversos , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/imunologiaRESUMO
A recent study reported an increase of brain tissue copper content in the lentiform nuclei of patients with primary adult-onset dystonia. In this study we analyze copper-metabolizing proteins (Menkes protein, Wilson protein, ceruloplasmin) by Western blot analysis in frozen brain tissue (lentiform nuclei) of 3 patients with primary dystonia. Menkes protein was reduced in all patients, while Wilson protein and ceruloplasmin were increased in the 2 patients with focal dystonia and reduced in the patient with generalized dystonia. Our data provides further evidence for a disturbance of copper metabolism in primary dystonia.
Assuntos
Adenosina Trifosfatases/metabolismo , Proteínas de Transporte/metabolismo , Proteínas de Transporte de Cátions , Ceruloplasmina/metabolismo , Distúrbios Distônicos/metabolismo , Globo Pálido/metabolismo , Putamen/metabolismo , Proteínas Recombinantes de Fusão , Adenosina Trifosfatases/análise , Idoso , Proteínas de Transporte/análise , Ceruloplasmina/análise , Cobre/metabolismo , ATPases Transportadoras de Cobre , Feminino , Globo Pálido/química , Humanos , Masculino , Pessoa de Meia-Idade , Putamen/química , Valores de Referência , População BrancaRESUMO
In EAN, TCR variable (V) gene usage is still controversial. A dominant usage of a TCR Vbeta4-associated idiotype has been reported. To assess the role of TCR Vbeta4 positive T-cells in susceptibility to induction of EAN, we suppressed the selection of this idiotype by neonatal treatment of Lewis rats with anti-TCR Vbeta4 monoclonal antibody (mAb). Anti-Vbeta4 treatment had no effect on development of clinical disease after immunization with the neuritogenic P2-peptide amino acids (aa) 53-78. Furthermore, lymph node cells from anti-Vbeta4 treated animals isolated after immunization with P2-peptide did not exhibit a reduced proliferative response towards whole P2-protein or P2-peptide. Our results indicate that T-cells utilizing other TCR V chains can functionally replace the neuritogenic cell population, which is dominant in stable T-cell lines.
Assuntos
Neurite Autoimune Experimental/etiologia , Receptores de Antígenos de Linfócitos T alfa-beta/fisiologia , Animais , Anticorpos Monoclonais/imunologia , Células Cultivadas , Feminino , Imunização , Depleção Linfocítica , Ratos , Ratos Endogâmicos LewRESUMO
In experimental autoimmune neuritis (EAN), T-cell receptor (TCR) variable (V)-region gene usage by neuritogenic T cells has been reported to be clonally restricted at the RNA level. This study was designed to verify TCR usage by neuritogenic T cells at the protein level. We generated two monoclonal antibodies (mAbs) 7H4 and 8G8 specific for a Vbeta4/Valpha11 associated idiotype expressed by the majority of neuritogenic cells of P2-specific T-cell lines. The remaining neuritogenic P2-specific T cells either exhibited a dominant usage of the TCR Vbeta13 chain recognized by the recently generated mAbs 17D5 and 18B1 or showed diverse Vbeta usage. Treatment of adoptive-transfer (AT)-EAN or of EAN actively induced with the neuritogenic P2 peptide by mAbs 7H4 and 8G8 led to a partial, but significant, reduction of clinical disease. Treatment with Vbeta13-specific mAb 17D5 had no clear effect on active EAN. Our data show that at least three different TCR are used by P2-specific pathogenic T cells in EAN, an animal model for human inflammatory neuropathies.
Assuntos
Genes Codificadores dos Receptores de Linfócitos T , Neurite Autoimune Experimental/genética , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Transferência Adotiva , Sequência de Aminoácidos , Animais , Anticorpos , Anticorpos Monoclonais/farmacologia , Especificidade de Anticorpos , Linhagem Celular , Epitopos , Feminino , Heterogeneidade Genética , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Neurite Autoimune Experimental/imunologia , Neurite Autoimune Experimental/terapia , Testes de Precipitina , RNA/genética , RNA/isolamento & purificação , Ratos , Ratos Endogâmicos Lew , Receptores de Antígenos de Linfócitos T alfa-beta/imunologia , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Wallerian degeneration, induced after injury to a peripheral nerve, is associated with upregulation of proinflammatory cytokines, which are suggested to contribute to the development of lesion-induced neuropathic pain. In chronic constrictive injury (CCI), an animal model of injury-induced painful mononeuropathy, inhibition of synthesis, release, or function of the cytokine tumor necrosis factor-alpha (TNF) results in reduced pain-associated behavior. Here, changes of TNF content in rat sciatic nerves after CCI (days 0, 0.5, 1, 3, 7 and 14) were investigated by enzyme-linked-immunoassay. Low levels of TNF were already detectable in control nerves. Concentrations increased rapidly after CCI, with a maximum (2.7-fold) at 12 h, and remained elevated on a lower level until day 3. Baseline levels were reached again at day 14. These results indicate that TNF is produced at an early time point in the cascade of events resulting in Wallerian degeneration and hyperalgesia following peripheral nerve injury. Given that only prophylactic treatment with TNF inhibitors efficiently reduces hyperalgesia in CCI, TNF seems to contribute to the initiation of neuropathic pain in this model.
Assuntos
Hiperalgesia/fisiopatologia , Regeneração Nervosa/fisiologia , Proteínas do Tecido Nervoso/biossíntese , Nervo Isquiático/lesões , Fator de Necrose Tumoral alfa/análise , Degeneração Walleriana , Animais , Constrição , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Feminino , Ratos , Ratos Sprague-Dawley , Nervo Isquiático/metabolismo , Nervo Isquiático/fisiologia , Sensibilidade e Especificidade , Fatores de Tempo , Fator de Necrose Tumoral alfa/biossínteseRESUMO
Guillain-Barré syndrome (GBS) is often associated with serum antibodies to glycoconjugates such as GM1 and GQ1b. The pathogenic role of these antibodies and other serum factors has not yet been clarified. We have investigated the effect of serum, plasma filtrate, and highly purified IgG and IgM from 10 patients with typical GBS on motor nerve terminals in the mouse hemidiaphragm. Quantal endplate currents were recorded by means of a perfused macro-patch-clamp electrode. The plasma filtrate of all GBS patients led to a 5- to 20-fold reduction of evoked quantal release within 7 to 15 minutes of continuous superfusion. In 4 patients, the amplitudes of single quanta were clearly reduced (by 10-66% of control values), indicating an additional postsynaptic action. Blocking effects could be reversed to a variable degree within 15 to 18 minutes after washout. Purified IgG was as effective as native serum, whereas a purified GBS IgM fraction did not block transmission. Sera from convalescent patients and IgG from healthy subjects were without blocking effect. The effects were complement independent and there was no link to the presence (in 6 patients) or absence (in 4 patients) of detectable antibodies to GM1 or GQ1b. In GBS, antibodies to an undetermined antigen depress the presynaptic transmitter release and, in some cases, the activation of postsynaptic channels. We suggest that weakness in the acute stage of GBS may be caused in part by circulating antibodies.
Assuntos
Anticorpos/farmacologia , Imunoglobulina G/farmacologia , Bloqueadores Neuromusculares/farmacologia , Polirradiculoneuropatia/imunologia , Adulto , Idoso , Animais , Fenômenos Fisiológicos Sanguíneos , Eletrofisiologia , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Pessoa de Meia-Idade , Terminações Nervosas/fisiologia , Junção Neuromuscular/efeitos dos fármacos , Técnicas de Patch-Clamp , Polirradiculoneuropatia/sangue , Sinapses/fisiologia , Transmissão Sináptica/efeitos dos fármacosRESUMO
Miller-Fisher syndrome, a variant of an acute inflammatory neuropathy is often associated with serum antibodies to the ganglioside GQ1b, but the pathogenic role of these antibodies and other serum factors is unclear. We here investigated the effect of highly purified immunoglobulin G (IgG) from patients with typical Miller-Fisher syndrome, recording quantal endplate currents by means of a perfused macro-patch-clamp electrode on hemidiaphragms of adult mice. The GQ1b-positive and the GQ1b-negative Miller-Fisher IgG as well as its monovalent Fab-fragments depressed evoked quantal release in a fast and fully reversible, concentration and voltage dependent manner. The time-course of quantal release was changed with the late releases becoming more frequent. The extent of depression of release followed a Michaelis-Menten kinetic and depended on the extracellular calcium concentration. In addition the amplitude of quanta was reduced postsynaptically. IgG and sera from healthy subjects had no effect. Our results indicate that in Miller-Fisher syndrome, IgG antibodies to an undetermined antigen depress the release process, most likely by interfering with the presynaptic Ca2+ inflow or by interacting with proteins of the exocytotic apparatus, and prevent the activation of postsynaptic channels. Antibodies thus seem to be one pathogenic factor for muscle weakness in Miller-Fisher syndrome and our findings may explain why muscle strength recovers rapidly after therapeutical plasmapheresis.
Assuntos
Imunoglobulina G/farmacologia , Síndrome de Miller Fisher/imunologia , Neurônios Motores/fisiologia , Junção Neuromuscular/imunologia , Transmissão Sináptica/imunologia , Animais , Reações Antígeno-Anticorpo , Autoanticorpos/farmacologia , Cálcio/metabolismo , Fragmentos Fab das Imunoglobulinas/farmacologia , Potenciais da Membrana/fisiologia , Camundongos , Camundongos Endogâmicos BALB C , Junção Neuromuscular/metabolismo , Técnicas de Patch-Clamp , Terminações Pré-Sinápticas/imunologia , Terminações Pré-Sinápticas/metabolismo , Fatores de TempoRESUMO
Exposure of T cells to their specific antigen normally results in proliferation, but in the presence of high and repeatedly administered doses of antigen, T cells may undergo apoptosis. Here we demonstrate that i.v. administration of as little as 100 microg of recombinant P2 protein twice daily completely prevents experimental autoimmune neuritis induced by adoptive transfer of neuritogenic P2-specific T cells or by immunization with the neuritogenic P2-peptide-spanning amino acids 53-78. Antigen treatment started after disease onset markedly ameliorated experimental autoimmune neuritis. The mechanism of action may be through programmed T cell death; a profound increase of the rate of apoptosis was seen in inflammatory foci of peripheral nerves and in the spleen. There was no cytokine switch by our Th1 cells after exposure to their specific antigen, but increased secretion of interferon gamma and tumor necrosis factor alpha was demonstrated. High antigen dose therapy using recombinant, pathogen-free protein may prove useful for the treatment of autoimmune inflammatory disorders of the nervous system.
Assuntos
Apoptose , Imunização , Proteína Básica da Mielina/uso terapêutico , Proteína P2 de Mielina , Neurite Autoimune Experimental/terapia , Fragmentos de Peptídeos/uso terapêutico , Linfócitos T/imunologia , Transferência Adotiva , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Feminino , Humanos , Inflamação/terapia , Ativação Linfocitária/efeitos dos fármacos , Proteína Básica da Mielina/genética , Proteína Básica da Mielina/imunologia , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/imunologia , Ratos , Ratos Endogâmicos Lew , Proteínas Recombinantes/uso terapêuticoRESUMO
The major neuritogenic protein of peripheral nerve myelin is the P2 protein. Human P2 is a candidate autoantigen in inflammatory demyelinating diseases of the peripheral nervous system. Since human P2 is not readily available, we produced full-length recombinant human P2 protein (rhP2) in Escherichia coli. RhP2 was recognized by neuritogenic rat T cell lines and induced experimental autoimmune neuritis in Lewis rats. Production of rhP2 allowed the generation of human T cell lines reactive to the autologous protein. Studies of human T cell autoreactivity as well as efforts to use hP2 as a tolerogen will be facilitated by the large-scale expression of rhP2.
Assuntos
Proteína P2 de Mielina/imunologia , Linfócitos T/imunologia , Sequência de Aminoácidos , Animais , Autoanticorpos , Sequência de Bases , Escherichia coli/genética , Feminino , Humanos , Imunização , Dados de Sequência Molecular , Proteína P2 de Mielina/isolamento & purificação , Neurite (Inflamação)/imunologia , Sistema Nervoso Periférico/imunologia , Ratos , Ratos Endogâmicos Lew , Proteínas Recombinantes/imunologiaRESUMO
A neuromuscular blocking factor has been described in the serum of patients with Miller-Fisher syndrome (MFS). We here examined the effect of immunoglobulins (Ig) on neuromuscular transmission in mice recording quantal endplate currents by means of a perfused macro-patch-clamp electrode. Ig and IgM- and IgG-fractions from an anti-GQ1b-positive patient with typical MFS were highly purified. After application of MFS-IgG, quantal release decreased 1000-fold within 2 min. Returning to control solution the average release came back to the baseline level within 4 min. In contrast, control-IgG and MFS-IgM did not cause any blocking effect. The very fast and fully reversible presynaptic blockade of release caused by the highly purified IgG-fraction may be one factor producing muscle weakness in MFS.
Assuntos
Doenças Desmielinizantes/imunologia , Imunoglobulina G/imunologia , Junção Neuromuscular/fisiologia , Doenças do Sistema Nervoso Periférico/imunologia , Animais , Diafragma/efeitos dos fármacos , Diafragma/fisiologia , Humanos , Imunoglobulina M/imunologia , Técnicas In Vitro , Camundongos , Camundongos Endogâmicos BALB C , Placa Motora/efeitos dos fármacos , Síndrome , Tetrodotoxina/farmacologiaRESUMO
Se presenta un caso de ileo mecánico por invaginación intestinal, cuyo origen resulta ser una malformación congénita con las características de divertículo ileocecal, desarrollado dentro de la pared del ciego, que se resuelve quirúrgicamente mediante la resección del segmento afectado