RESUMO
We propose an approach to determine the excitation energy distribution due to multiphoton absorption in the case of excited systems following decays to produce different ion species. This approach is based on the measurement of the time-resolved photoion position spectrum by using velocity map imaging spectrometry and an unfocused laser beam with a low fluence and homogeneous profile. Such a measurement allows us to identify the species and the origin of each ion detected and to depict the energy distribution using a pure Poisson's equation involving only one variable which is proportional to the absolute photon absorption cross section. A cascade decay model is used to build direct connections between the energy distribution and the probability to detect each ionic species. Comparison between experiments and simulations permits the energy distribution and accordingly the absolute photon absorption cross section to be determined. This approach is illustrated using C60 as an example. It may therefore be extended to a wide variety of molecules and clusters having decay mechanisms similar to those of fullerene molecules.
RESUMO
BACKGROUND: Cognitive dysfunction is frequently seen in neuromyelitis optica (NMO). However, the features and influencing factors of cognitive impairment of Chinese NMO patients are unclear. OBJECTIVE: To investigate the patterns of cognitive impairment in Chinese NMO patients, and correlate the neuropsychiatric scores with clinical and MRI parameters. METHODS: Thirty-six Chinese NMO patients, and 30 sex and age-matched healthy controls were recruited with extensive neuropsychological assessments, using the modified Minimal Assessment of Cognitive Function in MS (MACFIMS). The demographic and clinical characteristics as well as MRI parameters were compared between cognitively impaired (CI) and cognitively preserved (CP) patients. RESULTS: NMO patients were significantly impaired in the Paced Auditory Serial Addition Task (P<0.05), the Symbol Digit Modalities Test (P<0.001), the California Verbal Learning Test-Second Edition (P<0.05), the Brief Visuospatial Memory Test-Revised (P<0.05) and semantic fluency (P<0.001). Only lower education level was associated with cognitive dysfunction in NMO (odds ratio: 0.57, P<0.05). There were no significant differences of MRI parameters regarding white matter (WM) lesions, grey matter and WM brain volume between CI and CP patients. CONCLUSIONS: Chinese NMO patients particularly demonstrated cognitive impairment in information processing speed, executive function and memory. Lower education level was the main factor contributing to cognitive impairment in NMO.
Assuntos
Transtornos Cognitivos/etiologia , Função Executiva/fisiologia , Transtornos da Memória/etiologia , Neuromielite Óptica/complicações , Desempenho Psicomotor/fisiologia , Adulto , China , Transtornos Cognitivos/diagnóstico , Escolaridade , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Transtornos da Memória/diagnóstico , Pessoa de Meia-Idade , Neuromielite Óptica/diagnósticoRESUMO
The differentiation and maintenance of Th17 cells require a unique cytokine milieu and activation of lineage-specific transcription factors. This process appears to be antagonized by the transcription factor T-bet, which controls the differentiation of Th1 cells. Considering that T-bet-deficient (T-bet(-/-) ) mice are largely devoid of natural killer (NK) cells due to a defect in the terminal maturation of these cells, and because NK cells can influence the differentiation of T helper cells, we investigated whether the absence of NK cells in T-bet-deficient mice contributes to the augmentation of autoreactive Th17 cell responses. We show that the loss of T-bet renders the transcription factors Rorc and STAT3 highly responsive to activation by stimuli provided by NK cells. Furthermore, reconstitution of T-bet(-/-) mice with wild-type NK cells inhibited the development of autoreactive Th17 cells through NK cell-derived production of IFN-γ. These results identify NK cells as critical regulators in the development of autoreactive Th17 cells and Th17-mediated pathology.
Assuntos
Células Matadoras Naturais/imunologia , Proteínas com Domínio T/metabolismo , Células Th17/imunologia , Animais , Autoantígenos/imunologia , Comunicação Celular/genética , Comunicação Celular/imunologia , Diferenciação Celular/genética , Diferenciação Celular/imunologia , Células Cultivadas , Citocinas/imunologia , Citotoxicidade Imunológica , Regulação da Expressão Gênica/genética , Regulação da Expressão Gênica/imunologia , Tolerância Imunológica , Células Matadoras Naturais/transplante , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/genética , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/metabolismo , Fator de Transcrição STAT3/genética , Fator de Transcrição STAT3/metabolismo , Proteínas com Domínio T/genética , Células Th1/imunologiaRESUMO
BACKGROUND AND PURPOSE: The cerebellum plays an important role in motor and cognitive functions. However, whether and how the normal-appearing cerebellum is impaired in patients with neuromyelitis optica spectrum disorders remain unknown. We aimed to identify the occult structural damage of the cerebellum in neuromyelitis optica spectrum disorder and its possible causes at the level of substructures. MATERIALS AND METHODS: Normal-appearing gray matter volume of the cerebellar lobules and nuclei and normal-appearing white matter volume of the cerebellar peduncles were compared between patients with neuromyelitis optica spectrum disorder and healthy controls. RESULTS: The cerebellar damage of patients with neuromyelitis optica spectrum disorder in the hemispheric lobule VI, vermis lobule VI, and all cerebellar nuclei and peduncles was related only to spinal lesions; and cerebellar damage in the hemispheric lobules VIII and X was related only to the aquaporin-4 antibody. The mixed cerebellar damage in the hemispheric lobules V and IX and vermis lobule Crus I was related mainly to spinal lesions; and mixed cerebellar damage in the hemispheric lobule VIIb was related mainly to the aquaporin-4 antibody. Other cerebellar substructures showed no significant cerebellar damage. CONCLUSIONS: We have shown that the damage in cerebellar normal-appearing white matter and normal-appearing gray matter is associated with aquaporin-4-mediated primary damage or axonal degeneration secondary to spinal lesions or both. The etiologic classifications of substructure-specific occult cerebellar damage may facilitate developing neuroimaging markers for assessing the severity and the results of therapy of neuromyelitis optica spectrum disorder occult cerebellar damage.
Assuntos
Cerebelo/patologia , Neuromielite Óptica/patologia , Adulto , Aquaporina 4/imunologia , Autoanticorpos/sangue , Feminino , Humanos , Imageamento por Ressonância Magnética/métodos , Masculino , Pessoa de Meia-Idade , Neuroimagem/métodos , Neuromielite Óptica/sangue , Neuromielite Óptica/imunologia , Medula Espinal/patologiaRESUMO
Experimental autoimmune encephalomyelitis (EAE) is a primary animal model of multiple sclerosis (MS). MS predominantly presents with evidence of lesions in the subcortical periventricular white matter regions of the brain. Research into the pathogenesis of the demyelinating lesions in the brain has been hampered by the fact that conventional models of EAE present with progressive ascending paralysis which recapitulates mainly the spinal cord lesions of multiple sclerosis. There is little evidence of brain involvement. Systemic administration of pertussis toxin (PTx) has been shown to induce the proinflammatory cascade of TGF-beta, IL-6, and Th17 in the central nervous system, which recently has been identified as essential in the development of EAE. To determine whether intracerebroventricular (icv) administration of PTx would result in subcortical periventricular demyelinating lesions in the brain, we examined the effect in a MOG induced EAE model. We found that icv PTx induced subcortical periventricular brain lesions that resemble the pathologic demyelinating lesions of MS. Moreover, icv PTx induced Th17 infiltration and increased expression of cytokines IL-6 and TGF-beta. We thus generated a highly reproducible model with remarkable histological similarities to the predominant demyelinating brain lesions seen in MS.
Assuntos
Ventrículos Cerebrais/efeitos dos fármacos , Encefalomielite Autoimune Experimental/induzido quimicamente , Camundongos , Esclerose Múltipla/induzido quimicamente , Toxina Pertussis/toxicidade , Animais , Ventrículos Cerebrais/imunologia , Ventrículos Cerebrais/patologia , Encefalomielite Autoimune Experimental/imunologia , Encefalomielite Autoimune Experimental/patologia , Feminino , Interleucina-6/metabolismo , Leucócitos/imunologia , Meningite/imunologia , Meningite/patologia , Camundongos Endogâmicos C57BL , Esclerose Múltipla/imunologia , Esclerose Múltipla/patologia , Toxina Pertussis/administração & dosagem , Fator de Crescimento Transformador beta/metabolismoRESUMO
The homeostasis of CD4+ CD25+ regulatory T cells (Tregs) depends on the cytokine interleukin (IL)-2. As IL-21 shares sequence homology with IL-2 and the IL-21 receptors contain a gamma-chain common to IL-2, we hypothesized that IL-21 could also affect the homeostasis of Tregs. We tested this hypothesis in experimental autoimmune encephalomyelitis (EAE), an animal model of relapsing-remitting human multiple sclerosis. We show that blockade of IL-21 in SJL/J mice before and after the induction of EAE enhances the influx of inflammatory cells into the central nervous system (CNS). The blockade of IL-21 leads to proliferation of proteolipid peptide (PLP(139-151))-autoreactive CD4+ T cells, which are capable to cause severe EAE in adoptively transferred recipient mice. Conversely, Tregs from mice where IL-21 was blocked, lose their capacity to prevent EAE induced PLP(139-151)-reactive T cells. Notably, direct effects of IL-21 on Tregs are confirmed by studies of blockade of IL-21 in mice expressing a green fluorescent protein 'knocked' into a Foxp3 allele, in which a reduction of the number of Tregs and a downregulation of their frequency and expression of Foxp3 are observed. These data suggest a role of the IL-21/IL-21R axis in the homeostasis of Tregs in CNS autoimmunity.
Assuntos
Encefalomielite Autoimune Experimental/imunologia , Homeostase/imunologia , Interleucinas/fisiologia , Linfócitos T Reguladores/imunologia , Sequência de Aminoácidos , Animais , Encefalomielite Autoimune Experimental/metabolismo , Feminino , Homeostase/genética , Humanos , Fragmentos Fc das Imunoglobulinas/fisiologia , Interleucinas/antagonistas & inibidores , Interleucinas/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Dados de Sequência Molecular , Receptores de Interleucina-21/biossíntese , Receptores de Interleucina-21/genética , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Linfócitos T Reguladores/metabolismoRESUMO
BACKGROUND AND PURPOSE: Whether gray matter impairment occurs in neuromyelitis optica is a matter of ongoing debate, and the association of gray matter impairment with cognitive deficits remains largely unknown. The purpose of this study was to investigate gray matter volume reductions and their association with cognitive decline in patients with neuromyelitis optica. MATERIALS AND METHODS: This study included 50 patients with neuromyelitis optica and 50 sex-, age-, handedness-, and education-matched healthy subjects who underwent high-resolution structural MR imaging examinations and a battery of cognitive assessments. Gray matter volume and cognitive differences were compared between the 2 groups. The correlations of the regional gray matter volume with cognitive scores and clinical variables were explored in the patients with neuromyelitis optica. RESULTS: Compared with healthy controls (635.9 ± 51.18 mL), patients with neuromyelitis optica (602.8 ± 51.03 mL) had a 5.21% decrease in the mean gray matter volume of the whole brain (P < .001). The significant gray matter volume reduction in neuromyelitis optica affected the frontal and temporal cortices and the right thalamus (false discovery rate correction, P < .05). The regional gray matter volumes in the frontal and temporal cortices were negatively correlated with disease severity in patients with neuromyelitis optica (Alphasim correction, P < .05). Patients with neuromyelitis optica had impairments in memory, information processing speed, and verbal fluency (P < .05), which were correlated with gray matter volume reductions in the medial prefrontal cortex and thalamus (Alphasim correction, P < .05). CONCLUSIONS: Gray matter volume reduction is present in patients with neuromyelitis optica and is associated with cognitive impairment and disease severity in this group.
Assuntos
Encéfalo/patologia , Transtornos Cognitivos/diagnóstico , Transtornos Cognitivos/patologia , Substância Cinzenta/patologia , Aumento da Imagem , Imageamento por Ressonância Magnética , Neuromielite Óptica/diagnóstico , Neuromielite Óptica/patologia , Adulto , Atrofia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Testes Neuropsicológicos , Tamanho do Órgão/fisiologia , Córtex Pré-Frontal/patologia , Valores de Referência , Estatística como Assunto , Tálamo/patologiaRESUMO
Experimental autoimmune neuritis (EAN) is a T-cell mediated autoimmune disease of the peripheral nervous system, in which macrophages and T-cells feature prominently in nerve lesions. EAN represents a counterpart to Guillain-Barré syndrome in humans. In the present study, we investigated the in vitro and in vivo effects of Linomide (LS-2616, quinoline-3-carboxamide), a synthetic immunomodulatory compound, on macrophages in relation to EAN. Linomide strongly suppressed IFN-gamma and lipopolysaccharide (LPS)-induced IL-1 beta, TNF-alpha and IL-6 mRNA expression in macrophages in vitro as demonstrated by in situ hybridisation. Linomide administered daily subcutaneously from the day of inoculation completely prevented the development of clinical symptoms of EAN. Linomide administered from day 9 post immunisation (p.i.) significantly suppressed clinical EAN. Macrophages from Linomide-treated EAN rats showed decreased IL-1 beta, TNF-alpha and IL-6 mRNA expression in response to IFN-gamma and LPS. LPS-induced nitric oxide production by macrophages was also suppressed by Linomide in vitro. Linomide, however, does not affect macrophage death and release of lactate dehydrogenase. We conclude that Linomide may exert its actions in EAN and perhaps also in other autoimmune disease models, by suppressing macrophage functions.
Assuntos
Adjuvantes Imunológicos/farmacologia , Hidroxiquinolinas/farmacologia , Macrófagos/efeitos dos fármacos , Neurite Autoimune Experimental/prevenção & controle , Animais , Regulação para Baixo , Interleucina-1/genética , Interleucina-6/genética , Lipopolissacarídeos/farmacologia , Macrófagos/fisiologia , Masculino , Óxido Nítrico/biossíntese , RNA Mensageiro/análise , Ratos , Ratos Endogâmicos Lew , Fator de Necrose Tumoral alfa/genéticaRESUMO
Suppressive effects of the synthetic immunomodulatory drug Linomide have been shown in several autoimmune models, including antibody-mediated experimental autoimmune myasthenia gravis (EAMG), a model for human myasthenia gravis (MG). To define the mechanisms underlying EAMG suppression, we injected Linomide subcutaneously at different doses into Lewis rats immunized with Torpedo acetylcholine receptor (AChR) in complete Freund's adjuvant (CFA), and investigated AChR-specific T and B cell responses, and the levels of lymph node cells expressing mRNA of different cytokines after AChR stimulation in vitro. Both 160 and 16, but not 1.6, mg/kg/day of Linomide effectively suppressed clinical muscle weakness, accompanied by decreased AChR-induced T and B cell responses. Linomide also suppressed the mRNA expression of the Th1 cytokines IFN-gamma, IL-12 and TNF-alpha as well as the Th2 cytokines IL-4 and IL-10, which are important in the immunopathogenesis of EAMG by promoting antibody production. There were no differences for IL-1beta, IL-6, lymphotoxin or TGF-beta expression in Linomide-treated vs nontreated control EAMG rats. We conclude that Linomide suppresses clinical EAMG as well as B and T cell responses to AChR by counteracting the production of AChR-induced Th1 and Th2 cytokines.
Assuntos
Adjuvantes Imunológicos/farmacologia , Citocinas/antagonistas & inibidores , Hidroxiquinolinas/farmacologia , Miastenia Gravis/metabolismo , Animais , Anticorpos/análise , Divisão Celular , Citocinas/genética , Feminino , Imunoglobulina G/análise , Linfócitos/patologia , Monócitos/imunologia , Debilidade Muscular , Miastenia Gravis/patologia , Miastenia Gravis/fisiopatologia , RNA Mensageiro/metabolismo , Ratos , Ratos Endogâmicos Lew , Receptores Colinérgicos/imunologiaRESUMO
This study explores nasal administration of myelin basic protein (MBP) as a potential means of inducing tolerance to relapsing experimental autoimmune encephalomyelitis (PR-EAE), an experimental multiple sclerosis (MS) model that was induced in DA rats by immunization with rat spinal cord homogenate and incomplete Freund's adjuvant. DA rats received a total dosage of 0, 6, 60, 600 micrograms/rat of bovine MBP on ten consecutive days prior to immunization. EAE with typical course was observed in control rats receiving only PBS nasally, and in rats receiving 6 micrograms/rat of MBP. Rats receiving 60 micrograms/rat of MBP developed acute EAE but no relapse during 60 days of observation post immunization (p.i.). Only one of eight rats receiving 600 micrograms/rat of MBP developed slight, transient EAE. This protection was confirmed at the histology level and was associated with decreased levels of MBP-reactive IFN-gamma secreting Th1-like spleen cells on day 13 and 60 p.i. Rats receiving 60 and 600 micrograms/rat of MBP showed decreased serum anti-MBP IgG2b antibody levels on day 60 p.i., and rats receiving 600 micrograms/rat of MBP had marginally increased anti-MBP IgG1 antibody levels in serum compared to control EAE rats. Cytokine mRNA profiles in central nervous system (CNS) and spleen mononuclear cells were evaluated. Dose-dependent reduction of TNF-alpha mRNA expression were observed both in CNS and in splenocytes. Increased IL-4 and TGF-beta mRNA expression were observed in CNS of low (6 micrograms/rat) and median (60 micrograms/rat) dose of MBP tolerized rats and in splenocytes of rats tolerized with 600 micrograms/rat of MBP. We conclude that nasal administration of MBP in DA rat prevents EAE induced by immunization with whole rat spinal cord homogenate that, besides MBP, contains multiple antigenic myelin proteins. A mechanism involving MBP-reactive regulatory cells expressing IL-4 and TGF-beta mRNA acts as part in the induction of this tolerance.
Assuntos
Encefalomielite Autoimune Experimental/prevenção & controle , Interleucina-4/genética , Ativação Linfocitária/genética , Proteína Básica da Mielina/administração & dosagem , RNA Mensageiro/biossíntese , Células Th1/imunologia , Fator de Crescimento Transformador beta/genética , Administração Intranasal , Animais , Especificidade de Anticorpos , Bovinos , Relação Dose-Resposta Imunológica , Encefalomielite Autoimune Experimental/genética , Encefalomielite Autoimune Experimental/imunologia , Epitopos/imunologia , Tolerância Imunológica , Imunoglobulina G/sangue , Isotipos de Imunoglobulinas/sangue , Imunossupressores/administração & dosagem , Interleucina-4/biossíntese , Ativação Linfocitária/efeitos dos fármacos , Proteína Básica da Mielina/imunologia , Ratos , Ratos Endogâmicos , Recidiva , Medula Espinal/imunologia , Medula Espinal/metabolismo , Baço/imunologia , Baço/metabolismo , Células Th1/efeitos dos fármacos , Células Th1/metabolismo , Fator de Crescimento Transformador beta/biossínteseRESUMO
Nasal administration of Torpedo acetylcholine receptor (AChR) to Lewis rats prior to induction of experimental autoimmune myasthenia gravis (EAMG) is highly efficient in prevention of clinical weakness, and suppression of AChR-specific T and B cell responses. To identify possible antigenic determinants within the receptor which can modulate EAMG and anti-AChR response, we evaluated the effects of nasal administration of alpha 61-76, alpha 100-116, alpha 146-162, delta 354-367, and alpha 261-277 of Torpedo AChR at different doses on the tolerance induction against EAMG irrespective if given at lower, the same or higher doses than whole Torpedo AChR protein, that was confirmed to be highly efficient as tolerogen to EAMG. None of these peptides, neither administrated alone nor in combination, induced tolerance to EAMG. Peptide administration did not affect the levels or affinities of anti-AChR antibodies when compared with non-tolerized control EAMG rats, while administration of whole AChR protein affected both variables. The results may indicate that the T and B cell heterogeneity of AChR epitopes makes it difficult to induce tolerance using synthetic peptide.
Assuntos
Tolerância Imunológica , Miastenia Gravis/imunologia , Cavidade Nasal/imunologia , Fragmentos de Peptídeos/imunologia , Receptores Colinérgicos/imunologia , Animais , Anticorpos/análise , Feminino , Tolerância Imunológica/imunologia , Imunoglobulina G/imunologia , Músculos/fisiopatologia , Miastenia Gravis/fisiopatologia , Fragmentos de Peptídeos/síntese química , Ratos , Ratos Endogâmicos Lew , Receptores Colinérgicos/química , TorpedoRESUMO
Oral tolerization with acetylcholine receptor (AChR) and myelin basic protein (MBP) prior to immunization with AChR+MBP+ complete Freund's adjuvant (CFA) alleviated clinical signs of experimental autoimmune myasthenia gravis (EAMG)+experimental allergic encephalomyelitis (EAE) and AChR- or MBP-specific T and B cell responses. Tolerance induced via the nasal route needs much less tolerogen and may still be as effective as oral tolerance induction. We now immunized Lewis rats with AChR+MBP+bovine peripheral nerve myelin (BPM)+CFA, which resulted in a multiphasic clinical picture with a combination of clinical signs of the EAMG+EAE+experimental allergic neuritis (EAN), accompanied by massive macrophage infiltrations in sections of muscle, spinal cord and sciatic nerve, and strong T and B cell responses to AChR, MBP and BPM in lymphoid organs. Nasal administration of microg doses of AChR+MBP+BPM prior to immunization with a mixture of these antigens+CFA effectively suppressed the incidence and severity of clinical disease, reduced macrophage infiltrations in sections of muscle, spinal cord and sciatic nerve, and down-regulated autoreactive T cell responses to the three antigens in lymphoid organs. Numbers of AChR-, MBP-, BPM-reactive Th1 type of cytokine interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha mRNA expression in lymph node cells were markedly suppressed, while transforming growth factor-beta (TGF-beta) mRNA expression was upregulated from nasally tolerized rats, suggesting an active suppression mechanism may act partly in the induction of tolerance. The results implicate the possibility to establish multiple autoantigen-based vaccination for the prevention of autoimmune diseases in humans.
Assuntos
Autoantígenos/administração & dosagem , Doenças Autoimunes/prevenção & controle , Imunossupressores/administração & dosagem , Administração Intranasal , Animais , Autoantígenos/imunologia , Doenças Autoimunes/imunologia , Bovinos , Encefalomielite Autoimune Experimental/imunologia , Encefalomielite Autoimune Experimental/metabolismo , Encefalomielite Autoimune Experimental/prevenção & controle , Epitopos/genética , Feminino , Tolerância Imunológica , Imunoglobulina G/biossíntese , Imuno-Histoquímica , Interferon gama/genética , Interleucina-10/genética , Leucócitos Mononucleares/metabolismo , Debilidade Muscular/imunologia , Debilidade Muscular/metabolismo , Miastenia Gravis/metabolismo , Miastenia Gravis/prevenção & controle , Proteína Básica da Mielina/metabolismo , Bainha de Mielina/imunologia , Neurite Autoimune Experimental/imunologia , Neurite Autoimune Experimental/metabolismo , Neurite Autoimune Experimental/prevenção & controle , RNA Mensageiro/biossíntese , Ratos , Ratos Endogâmicos Lew , Receptores Colinérgicos/metabolismo , Linfócitos T/metabolismo , Fator de Crescimento Transformador beta/genética , Fator de Necrose Tumoral alfa/genéticaRESUMO
In-situ hybridization with labeled oligonucleotide probes was applied to explore cytokine and chemokine mRNA expression in sections of striated muscle, the target organ in experimental autoimmune myasthenia gravis (EAMG), induced in Lewis rats by immunization with acetylcholine receptor (AChR) and complete Freund's adjuvant (CFA). A transient burst of TNF-alpha, IL-1beta and IL-6 mRNA expressing cells was detected during the early phase of EAMG. This cytokine pattern was related to muscular infiltration of macrophages. Levels of IL-4, IL-10, IFN-gamma, cytolysin and TGF-beta mRNA expressing cells were low and observed mainly during the early phase of EAMG. C-C chemokine RANTES, MCP, MIP-1alpha and MIP-2 mRNA expressing cells were not detected over the course of EAMG. The low and transient expression of cytokines in EAMG muscle tissues suggests that the immune effector responses are unlikely operated by infiltrating cells in muscle. Muscular infiltrations in EAMG are unlikely due to local accumulation of C-C chemokines.
Assuntos
Quimiocinas/genética , Citocinas/genética , Músculo Esquelético/metabolismo , Miastenia Gravis/metabolismo , RNA Mensageiro/metabolismo , Animais , Feminino , Imuno-Histoquímica , Hibridização In Situ , Músculo Esquelético/patologia , Miastenia Gravis/patologia , Miastenia Gravis/fisiopatologia , Ratos , Ratos Endogâmicos LewRESUMO
Myasthenia gravis (MG) and experimental autoimmune myasthenia gravis (EAMG) are caused by auto-antibodies against the nicotinic acetylcholine receptor (AChR) at the postsynaptic membrane. To evaluate the extent to which the humoral immune response against AChR operates in the pathogenesis of EAMG, we immunized B-cell knockout (microMT) and wild type C57BL/6 mice with AChR in complete Freund's adjuvant. The ability of AChR-primed lymph node cells to proliferate and secrete IFN-gamma in response to AChR and its dominant peptide alpha 146-162 were intact in microMT as in wild type mice. Similar levels of mRNA for IFN-gamma, IL-4 and IL-10 in AChR-reactive lymph node cells were detected in microMT and wild type mice. However, microMT mice had no detectable anti-AChR antibodies and never developed clinical EAMG. We conclude that B-cells are critically required for the genesis of clinical EAMG, but not for AChR-specific T-cell priming.
Assuntos
Linfócitos B/imunologia , Miastenia Gravis/imunologia , Animais , Autoanticorpos/análise , Autoanticorpos/imunologia , Doenças Autoimunes/imunologia , Linfócitos B/metabolismo , Contagem de Células , Divisão Celular/fisiologia , Citocinas/metabolismo , Ensaio de Imunoadsorção Enzimática , Imunoglobulina G/biossíntese , Interferon gama/metabolismo , Linfonodos/citologia , Linfócitos/imunologia , Linfócitos/metabolismo , Linfócitos/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptores Nicotínicos/imunologiaRESUMO
One of the major consequences of stroke is brain injury caused by glutamate-mediated excitotoxicity. Glutamate-mediated excitatory activities are partially driven by ß2-containing nicotinic acetylcholine receptors (ß2-nAChRs). In examining the role of ß2-nAChRs in cerebral ischemic injury, excitotoxicity and stroke outcome, we found that deficiency of ß2-nAChRs attenuated brain infarction and neurological deficit at 24 and 72 h after transient middle cerebral artery occlusion (MCAO). Genetic deletion of ß2-nAChRs associated with reduced terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL(+)) and cleaved caspase-3(+) cells after MCAO, together with a reduction of extracellular glutamate and oxygen-glucose deprivation-induced increase of excitatory inputs in cortical neurons. Pharmacologic pretreatment with a selective ß2-nAChRs antagonist reduced brain infarction, neurological deficit, and MCAO-induced glutamate release. These findings suggest that deficiency of ß2-nAChRs, also achievable by pharmacological blockade, can decrease brain infarction and improve the neurological status in ischemic stroke. The improved outcome is associated with reduced extracellular glutamate level and lower excitatory inputs into ischemic neurons, suggesting a reduction of glutamate-mediated excitotoxicity in the mechanisms of neuroprotection.
Assuntos
Lesões Encefálicas/etiologia , Lesões Encefálicas/genética , Infarto da Artéria Cerebral Média/complicações , Infarto da Artéria Cerebral Média/genética , Receptores Nicotínicos/deficiência , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/genética , Animais , Lesões Encefálicas/tratamento farmacológico , Células Cultivadas , Córtex Cerebral/citologia , Di-Hidro-beta-Eritroidina/farmacologia , Di-Hidro-beta-Eritroidina/uso terapêutico , Modelos Animais de Doenças , Glucose/deficiência , Hipóxia/fisiopatologia , L-Lactato Desidrogenase/metabolismo , Imageamento por Ressonância Magnética , Masculino , Camundongos , Camundongos Knockout , Exame Neurológico , Fármacos Neuroprotetores/farmacologia , Receptores Nicotínicos/genética , Fatores de TempoRESUMO
The potential of stem cells in regenerative medicine, developmental biology, and drug discovery has been well documented. For example, stem cells have the extraordinary ability of self-renewal, and also give rise to many specialized cells. It is clear that stem cell technology has revolutionized our understanding of modern biology and medicine and provided new insights into the mechanisms controlling basic cell biology and various diseases. Nicotinic acetylcholine receptors (nAChRs) are prototypical members of the ligand-gated ion channel super family of neurotransmitter receptors that play many critical roles in brain and body function. It has been demonstrated that in addition to mediation of classical excitatory neurotransmission at some loci and modulation of release of neurotransmitters in some cases, nAChRs also play important roles in influencing synaptic architecture and plasticity as well as neuronal survival/death. Recently, emerging lines of evidence have suggested that nAChRs express on stem cells, where they likely mediate crucial effects of cholinergic signaling on stem cell survival/apoptosis, proliferation, differentiation and maturation. In this review, we summarize current development in cholinergic modulations of stem cell survival/apoptosis, proliferation and differentiation in order to evaluate the impact of nAChRs in stem cell biology and pathology.
Assuntos
Apoptose/fisiologia , Diferenciação Celular/fisiologia , Receptores Nicotínicos/metabolismo , Células-Tronco/metabolismo , Animais , Sobrevivência Celular/fisiologia , Humanos , Células-Tronco/citologiaAssuntos
Adjuvantes Imunológicos/uso terapêutico , Citocinas/biossíntese , Hidroxiquinolinas/uso terapêutico , Miastenia Gravis/imunologia , Células Th1/imunologia , Células Th2/imunologia , Animais , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Miastenia Gravis/fisiopatologia , Miastenia Gravis/terapia , RNA Mensageiro/biossíntese , Ratos , Ratos Endogâmicos Lew , Células Th1/efeitos dos fármacos , Células Th2/efeitos dos fármacos , Fatores de Tempo , Transcrição Gênica/efeitos dos fármacosRESUMO
PLeptin is an adipocyte-derived hormone with pleiotropic effects on energy homeostasis, endocrine and reproductive functions, and immune responses. The multiple actions of leptin have led to the design and development of several leptin-based approaches to modulate the metabolic and endocrine status, to reduce inflammation, and to improve immune responses. Here, we review the current patents on leptin in different clinic applications.
Assuntos
Doenças do Sistema Imunitário/tratamento farmacológico , Leptina/uso terapêutico , Doenças Metabólicas/tratamento farmacológico , Animais , Biomimética , Terapia Genética , Humanos , Imunização , Infertilidade/tratamento farmacológico , Neoplasias/tratamento farmacológico , Patentes como AssuntoRESUMO
BACKGROUND: Although the concept that an initial course of immune-suppression facilitates subsequent immune-modulation (such as Th1 to Th2 deviation) is attractive for several autoimmune diseases, such a mechanism for serial-combination therapy has never been formally demonstrated. Recently, brief mitoxantrone induction-chemotherapy followed by immune-modulation with glatiramer acetate (GA) was significantly more effective at reducing multiple sclerosis disease activity than with GA alone. OBJECTIVE: To examine whether the benefit of initial immune suppression with mitoxantrone before GA treatment is associated with more efficient immune modulation. METHODS: IgG1/IgG4 GA-reactive antibody profiles, previously established as markers of GA-induced Th2 immune-deviation, were prospectively measured in vivo in patients treated with GA alone or with mitoxantrone induction therapy followed by GA. RESULTS: Significant and sustained increase in IgG4 antibodies (and the anticipated reversal of the IgG1/IgG4 ratio) was seen in patients treated with GA alone. Combination therapy resulted in lesser IgG4 induction (and no reversal of IgG1/IgG4 ratio). Thus, the enhanced efficacy of mitoxantrone-GA combination regimen was associated with decreased, rather than increased, efficiency of shifting the GA-reactive IgG1/IgG4 antibody profile. CONCLUSION: These results provide important insights into mechanisms of combination therapy and therapeutic strategies for autoimmune diseases.
Assuntos
Fatores Imunológicos/administração & dosagem , Imunossupressores/administração & dosagem , Mitoxantrona/administração & dosagem , Esclerose Múltipla/tratamento farmacológico , Peptídeos/administração & dosagem , Adolescente , Adulto , Linfócitos B/efeitos dos fármacos , Linfócitos B/imunologia , Células Cultivadas , Esquema de Medicação , Quimioterapia Combinada , Feminino , Acetato de Glatiramer , Humanos , Imunoglobulina G/sangue , Imunoglobulina G/efeitos dos fármacos , Imunoglobulina G/imunologia , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/imunologia , Estudos Prospectivos , Método Simples-Cego , Células Th1/efeitos dos fármacos , Células Th1/imunologia , Células Th2 , Fatores de Tempo , Resultado do Tratamento , Adulto JovemRESUMO
CD4(+)CD25(+) regulatory T cells (Tregs) are potent immunosuppressors that are pivotal in the maintenance of self-tolerance. The involvement of Tregs in therapies for immune-mediated diseases has been proposed, but direct supporting evidence is still lacking. While investigating mechanisms underlying the clinical benefits of glatiramer acetate (GA) in an animal model of multiple sclerosis (MS), i.e., experimental autoimmune encephalomyelitis (EAE), we recently demonstrated that GA can protect mice deficient in the Th(2) cytokines IL-4, IL-10 and IL-4/IL-10 from acquiring EAE, suggesting that mechanisms other than Th(2) cells may be responsible for the therapeutic effects of GA. Here we demonstrate that GA treatment boosts the expression of Foxp3 on Tregs during EAE. Furthermore, adoptive transfer of purified Tregs from GA-treated EAE mice is more effective in preventing EAE development than Tregs from untreated EAE controls. Thus, our current data provide evidence that Tregs may be the major contributor to GA's therapeutic action in EAE and, possibly, MS. Further mechanistic studies to reveal the molecular events linking GA with Tregs may optimize GA treatment and lead to the development of new, even more effective therapies that utilize this mechanism of action.