RESUMO
Gaucher and Fabry diseases are the most prevalent sphingolipidoses. Chronic inflammation is activated in those disorders, which could play a role in pathogenesis. Significant degrees of amelioration occur in patients upon introduction of specific therapies; however, restoration to complete health status is not always achieved. The idea of an adjunctive therapy that targets inflammation may be a suitable option for patients. PPS is a mixture of semisynthetic sulfated polyanions that have been shown to have anti-inflammatory effects in mucopolysaccharidosis type I and II patients and animal models of type I, IIIA and VI. We hypothesized PPS could be a useful adjunctive therapy to inflammation for Gaucher and Fabry diseases. The objective of this work is to analyze the in vitro effect of PPS on inflammatory cytokines in cellular models of Gaucher and Fabry diseases, and to study its effect in Gaucher disease associated in vitro bone alterations. Cultures of peripheral blood mononuclear cells from Fabry and Gaucher patients were exposed to PPS. The secretion of proinflammatory cytokines was significantly reduced. Peripheral blood cells exposed to PPS from Gaucher patients revealed a reduced tendency to differentiate to osteoclasts. Osteoblasts and osteocytes cell lines were incubated with an inhibitor of glucocerebrosidase, and conditioned media was harvested in order to analyze if those cells secrete factors that induce osteoclastogenesis. Conditioned media from this cell cultures exposed to PPS produced lower numbers of osteoclasts. We could demonstrate PPS is an effective molecule to reduce the production of proinflammatory cytokines in in vitro models of Fabry and Gaucher diseases. Moreover, it was effective at ameliorating bone alterations of in vitro models of Gaucher disease. These results serve as preclinical supportive data to start clinical trials in human patients to analyze the effect of PPS as a potential adjunctive therapy for Fabry and Gaucher diseases.
Assuntos
Doença de Fabry/tratamento farmacológico , Doença de Gaucher/tratamento farmacológico , Inflamação/tratamento farmacológico , Poliéster Sulfúrico de Pentosana/farmacologia , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Doença de Fabry/patologia , Doença de Gaucher/patologia , Humanos , Inflamação/patologia , Leucócitos Mononucleares/efeitos dos fármacos , Doenças por Armazenamento dos Lisossomos/tratamento farmacológico , Doenças por Armazenamento dos Lisossomos/patologia , Lisossomos/efeitos dos fármacos , Lisossomos/genética , Osteoblastos/efeitos dos fármacos , Osteoclastos/efeitos dos fármacos , Osteócitos/efeitos dos fármacosRESUMO
Gaucher disease (GD) is caused by mutations in the glucosylceramidase ß (GBA 1) gene that confer a deficient level of activity of glucocerebrosidase (GCase). This deficiency leads to the accumulation of the glycolipid glucocerebroside in the lysosomes of cells, mainly in the monocyte/macrophage lineage. Its mildest form is Type I GD, characterized by non-neuronopathic involvement. Bone compromise is the most disabling aspect of the Gaucher disease. However, the pathophysiological aspects of skeletal alterations are not yet fully understood. The bone tissue homeostasis is maintained by a balance between resorption of old bone by osteoclasts and new bone formation by osteoblasts. A central player in this balance is the osteocyte as it controls both processes. We studied the involvement of osteocytes in an in vitro chemical model of Gaucher disease. The osteocyte cell line MLO-Y4 was exposed to conduritol-ß-epoxide (CBE), an inhibitor of GCase, for a period of 7, 14 and 21 days. Conditioned media from CBE-treated osteocytes was found to induce osteoclast differentiation. GCase inhibition caused alterations in Cx43 expression and distribution pattern and an increase in osteocyte apoptosis. Osteoclast differentiation involved osteocyte apoptotic bodies, receptor activator of nuclear factor κ-B ligand (RANKL) and soluble factors. Thus, our results indicate that osteocytes may have a role to play in the bone pathophysiology of GD.
Assuntos
Doença de Gaucher/patologia , Modelos Biológicos , Osteoclastos/patologia , Osteócitos/patologia , Osteogênese/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Células da Medula Óssea/patologia , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Conexina 43/metabolismo , Meios de Cultivo Condicionados/farmacologia , Feminino , Inositol/análogos & derivados , Inositol/farmacologia , Cadeias beta de Integrinas/metabolismo , Interleucina-6/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Macrófagos/patologia , Camundongos Endogâmicos C57BL , Osteoclastos/efeitos dos fármacos , Osteoclastos/metabolismo , Osteoprotegerina/metabolismo , Ligante RANK/farmacologia , SolubilidadeRESUMO
Pertussis has resurged during the last two decades in different countries. In particular in the 2010-2013 period large outbreaks were detected in US, Australia, UK and The Netherlands with significant mortality in infants. The epidemiological situation of pertussis points out the need to develop new vaccines and in this regard we previously developed a new vaccine based on outer membrane vesicles (OMVs) which have been shown to be safe and to induce protection in mice. Here we have further investigated the properties of OMVs vaccines; in particular we studied the contribution of pertussis toxin (PTx) and pertactin (Prn) in OMVs-mediated protection against pertussis. PTx-deficient OMVs and Prn-deficient OMVs were obtained from defective Bordetella pertussis mutants. The absence of PTx or Prn did compromise the protective capacity of the OMVs formulated as Tdap vaccine. Whereas the protective efficacy of the PTx-deficient OMVs in mice was comparable to Prn-deficient OMVs, the protective capacity of both of them was significantly impaired when it was compared with the wild type OMVs. Interestingly, using OMVs obtained from a B. pertussis strain which does not express any of the virulence factors but expresses the avirulent phenotype; we observed that the protective ability of such OMVs was lower than that of OMVs obtained from virulent B. pertussis phase. However, it was surprising that although the protective capacity of avirulent OMVs was lower, they were still protective in the used mice model. These results allow us to hypothesize that OMVs from avirulent phase shares protective components with all OMVs assayed. Using an immune proteomic strategy we identified some common components that could play an important role in protection against pertussis.
Assuntos
Proteínas da Membrana Bacteriana Externa/imunologia , Toxina Pertussis/imunologia , Vacina contra Coqueluche/imunologia , Fatores de Virulência de Bordetella/imunologia , Coqueluche/prevenção & controle , Animais , Antígenos de Bactérias/imunologia , Feminino , Camundongos Endogâmicos BALB CRESUMO
Despite high vaccination coverage rates, pertussis continues to be a global concern, with increased incidence widely noted. The current pertussis epidemiologic situation has been mainly attributed to waning immunity and pathogen adaptation. To improve the disease control, a new generation of vaccines capable to overcome those weaknesses associated to the current vaccines need to be developed. Previously we have demonstrated that the outer membrane vesicles obtained from the recombinant Bordetella pertussis strain expressing PagL enzyme (OMVs(BpPagL)) are good vaccine candidates to protect against pertussis. In this work the OMVs(BpPagL) formulated with diphtheria and tetanus toxoids (Tdap(OMVsBpPagL)) was used to evaluate its capacity to offer protection against Argentinean clinical isolates and to induce long-term immunity. To these aims BALB/c mice were immunized with Tdap(OMVsBpPagL) and challenged with sublethal doses of the clinical isolate Bp106 selected as a representative circulating isolate. Comparisons with a current commercial Tdap vaccine used at a dose in which pertussis toxin level was equivalent to that of Tdap(OMVsBpPagL) were performed. With the normalized doses of both vaccines we observed that Tdap(OMVsBpPagL) protected against the clinical isolate infection, whereas current commercial Tdap vaccine showed little protection against such pathogen. Regarding long-term immunity we observed that the Tdap(OMVsBpPagL) protective capacity against the recommended WHO reference strain persisted at least 9 months. In agreement with these results Tdap(OMVsBpPagL) induced Th1 and Th2 immune response. In contrast, commercial Tdap induced Th2 but weak Th1 responses. All results presented here showed that Tdap(OMVsBpPagL) is an interesting formulation to be considered for the development of novel acellular multi-antigen vaccine.
Assuntos
Proteínas da Membrana Bacteriana Externa/imunologia , Bordetella pertussis/classificação , Proteção Cruzada , Vacinas contra Difteria, Tétano e Coqueluche Acelular/imunologia , Coqueluche/prevenção & controle , Animais , Anticorpos Antibacterianos/sangue , Formação de Anticorpos , Bordetella pertussis/genética , Feminino , Genótipo , Memória Imunológica , Camundongos , Camundongos Endogâmicos BALB C , Toxina Pertussis/imunologia , Proteínas Recombinantes/imunologia , Células Th1/imunologia , Células Th2/imunologia , Vacinas Acelulares/imunologiaRESUMO
INTRODUCCION: La tos convulsa o pertussis es una enfermedad respiratoria que es más severa en los lactantes. Antes de que la vacunación infantil se introdujera en la década de 1950, la tos convulsa era una de las principales causas de mortalidad infantil en el mundo. La enfermedad hoy es reconocida como una infección frecuente, no sólo para los niños sino también para los adultos. Las razones de esta situación epidemiológica y las estrategias de control para la enfermedad son objeto de debate en la comunidad científica. En este contexto, los modelos matemáticos se utilizan cada vez más como un herramienta no sólo para el análisis, sino también para predicciones con el fin de contribuir al conocimiento de este complejo problema.OBJETIVO: En este estudio se presenta un modelo compartamentalizado, que de manera simplificada permite describir la propagación de la tos convulsa en la Argentina y evaluar el impacto de los cambios en el calendario de vacunación en el control de la enfermedad.METODOS: El modelo epidemiológico aplicado considera que la exposición a la tos convulsa a través de la infección natural o vacunación induce una respuesta inmune que previene la enfermedad grave. Además, se considera que estos efectos protectores son temporales debido a la disminución de la inmunidad.RESULTADOS: El estudio señala que la dosis administrada a los 11 años (recientemente introducidos en el esquema de vacunación de Argentina) disminuye la incidencia de la enfermedad en el grupo etario de 11 a 13 años de edad en una proporción de alrededor del 40%. Sin embargo, este refuerzo podría tener un impacto mucho menor (menos del 5%) para los niños menores de 1 año de edad que son el grupo más vulnerable.CONCLUSIONES: Nuestro estudio sugiere que un esfuerzo dirigido a la mejora de la cobertura de las primeras dosis tendrá un impacto mucho mayor que el refuerzo de los 11 en lo que se refiere a la reducción de la incidencia de tos convulsa en los niños más pequeños.
INTRODUCTION: Whooping cough, or pertussis, is a respiratory disease that is mose severe in infants. Before childhood vaccination was introduced in the 1950s, pertussis was a major cause of infant mortality worldwide. The disease is now recognized as a frequent infection not only for infants but also for adults. The reasons for this epidemiological situation and strategies for disease control are matters of debate in the scientific community. In this context, the mathematical models are being used increasingly as a tool not only for analysis but also for predictions in order to contribute to the knowledge of this complex problem.OBJECTIVE: The study has a compartmental model that, in a simplistic way, allows to describe the propagation of pertussis in Argentina and to assess the impact of changes in the vaccination schedule on the disease control.METHODS: The model here presented considers that pertussis exposure through natural infection or vaccination induces an immune response that prevents severe disease and assumes that these protective effects are temporary due to waning of immunity.RESULTS: The study points out that the dose given at 11 years of age (recently introduced in Argentica vaccination schedule) would decrease around 40% the incidence of the disease in the age group from 11 to 13 years old. However, this reinforcement would have a much lower impact (less than 5%) in children under 1 year, who are the most vulnerable group.CONCLUSIONS: It would be important to make an effort towards vigilance, so as to improve the coverage of the primary close and then significantly reduce the incidence of pertussis in the youngest children.
Assuntos
Lactente , Pré-Escolar , Criança , Adolescente , Coqueluche , Vacina contra Coqueluche , Vacinação em Massa , Argentina , Saúde PúblicaRESUMO
INTRODUCCION: La tos convulsa o pertussis es una enfermedad respiratoria que es más severa en los lactantes. Antes de que la vacunación infantil se introdujera en la década de 1950, la tos convulsa era una de las principales causas de mortalidad infantil en el mundo. La enfermedad hoy es reconocida como una infección frecuente, no sólo para los niños sino también para los adultos. Las razones de esta situación epidemiológica y las estrategias de control para la enfermedad son objeto de debate en la comunidad científica. En este contexto, los modelos matemáticos se utilizan cada vez más como un herramienta no sólo para el análisis, sino también para predicciones con el fin de contribuir al conocimiento de este complejo problema.OBJETIVO: En este estudio se presenta un modelo compartamentalizado, que de manera simplificada permite describir la propagación de la tos convulsa en la Argentina y evaluar el impacto de los cambios en el calendario de vacunación en el control de la enfermedad.METODOS: El modelo epidemiológico aplicado considera que la exposición a la tos convulsa a través de la infección natural o vacunación induce una respuesta inmune que previene la enfermedad grave. Además, se considera que estos efectos protectores son temporales debido a la disminución de la inmunidad.RESULTADOS: El estudio señala que la dosis administrada a los 11 años (recientemente introducidos en el esquema de vacunación de Argentina) disminuye la incidencia de la enfermedad en el grupo etario de 11 a 13 años de edad en una proporción de alrededor del 40%. Sin embargo, este refuerzo podría tener un impacto mucho menor (menos del 5%) para los niños menores de 1 año de edad que son el grupo más vulnerable.CONCLUSIONES: Nuestro estudio sugiere que un esfuerzo dirigido a la mejora de la cobertura de las primeras dosis tendrá un impacto mucho mayor que el refuerzo de los 11 en lo que se refiere a la reducción de la incidencia de tos convulsa en los niños más pequeños.
INTRODUCTION: Whooping cough, or pertussis, is a respiratory disease that is mose severe in infants. Before childhood vaccination was introduced in the 1950s, pertussis was a major cause of infant mortality worldwide. The disease is now recognized as a frequent infection not only for infants but also for adults. The reasons for this epidemiological situation and strategies for disease control are matters of debate in the scientific community. In this context, the mathematical models are being used increasingly as a tool not only for analysis but also for predictions in order to contribute to the knowledge of this complex problem.OBJECTIVE: The study has a compartmental model that, in a simplistic way, allows to describe the propagation of pertussis in Argentina and to assess the impact of changes in the vaccination schedule on the disease control.METHODS: The model here presented considers that pertussis exposure through natural infection or vaccination induces an immune response that prevents severe disease and assumes that these protective effects are temporary due to waning of immunity.RESULTS: The study points out that the dose given at 11 years of age (recently introduced in Argentica vaccination schedule) would decrease around 40% the incidence of the disease in the age group from 11 to 13 years old. However, this reinforcement would have a much lower impact (less than 5%) in children under 1 year, who are the most vulnerable group.CONCLUSIONS: It would be important to make an effort towards vigilance, so as to improve the coverage of the primary close and then significantly reduce the incidence of pertussis in the youngest children.