RESUMO
A family of platyhelminth tegument-specific proteins comprising of one or two calcium ion binding EF-hand and a dynein light chain-like domain, termed tegumental proteins, are considered as candidates of vaccine. In this study, we cloned and characterized SjTP22.4, a novel membrane-anchored tegumental protein in Schistosoma japonicum with theoretic MW of 22.4. The recombinant SjTP22.4 could be recognized by S. japonicum infected sera. Immunofluorescence revealed that this protein is not only located on the surface of tegument of adult and schistosomulum and cercaria, but also in the parenchymatous tissues and intestinal epithelium. Circular dichroism (CD) measurement demonstrated rSjTP22.4 had Ca(2+)-binding ability. The rSjTP22.4 vaccination without adjuvants produced comparable high level of antibody with that of immunization with adjuvants together indicated it was an antigen of strong antigenicity. The level of IgG1 is much higher than that of IgG2a and IgE is nearly negative in S. japonicum-infected and rSjTP22.4 immunized mice. In cercaria challenge experiment, mice vaccinated with SjTP22.4 showed no reduction in adult burden and egg production, comparing with the control mice, but 41% decrease in egg mature rate and 32% reduction in liver egg granuloma area. However, the SjTP22.4 immunized mice that received praziquantel treatment at 10d post infection caused 26% reduction in adult burden and 53% decrease in egg mature rate, comparing with the control mice only received praziquantel treatment. In conclusion, SjTP22.4 is a valuable vaccine candidate for S. japonicum of anti-pathogenesis and anti-transmission effect and plays a synergetic role in praziquantel to kill schistosomulum.
Assuntos
Proteínas de Ligação ao Cálcio/imunologia , Proteínas de Helminto/imunologia , Praziquantel/farmacologia , Schistosoma japonicum/imunologia , Esquistossomose Japônica/terapia , Adjuvantes Imunológicos/administração & dosagem , Animais , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/genética , Antígenos de Helmintos/imunologia , Proteínas de Ligação ao Cálcio/genética , Dicroísmo Circular , Clonagem Molecular , Sinergismo Farmacológico , Dineínas/genética , Dineínas/metabolismo , Feminino , Fertilidade , Imunofluorescência , Proteínas de Helminto/genética , Imunoglobulina G/sangue , Mucosa Intestinal/metabolismo , Estágios do Ciclo de Vida/efeitos dos fármacos , Estágios do Ciclo de Vida/imunologia , Fígado/imunologia , Fígado/parasitologia , Fígado/patologia , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/imunologia , Camundongos , Peso Molecular , Contagem de Ovos de Parasitas , Estrutura Terciária de Proteína , Proteínas Recombinantes/sangue , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Schistosoma japonicum/efeitos dos fármacos , Schistosoma japonicum/genética , Esquistossomose Japônica/imunologia , Esquistossomicidas/farmacologia , VacinaçãoRESUMO
To understand the defense mechanisms of Crustacean animals, brine shrimp Artemia sinica prophenoloxidase (AsproPO) cDNA was cloned and its expression at early developmental stages was examined by reverse-transcription PCR (RT-PCR) and semi-quantitative RT-PCR, and activity of phenoloxidase (PO) at different developmental stages was further detected by using l-3,4-dihydroxyphenylalanine (l-DOPA) as a specific substrate in this study. It was found that the full-length of AsproPO cDNA is 2125 bp and it contains an open reading frame of 2100 bp encoding a protein of 699 amino acids. The deduced amino acid sequence of AsproPO has two putative copper binding sites highly conserved in Arthropods. Semi-quantitative RT-PCR analyses showed that the gene of AsproPO expressed at Emergence, Instar I and Instar II stages but did not at 0 h and 6 h stages. Activity measurement showed that PO activity could only be detected at Instar II stage but the other measured stages. All these implied that Artemia proPO immune system was complexly modulated during early development.
Assuntos
Artemia/enzimologia , Artemia/imunologia , Catecol Oxidase/genética , Catecol Oxidase/metabolismo , Precursores Enzimáticos/genética , Precursores Enzimáticos/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/imunologia , Estágios do Ciclo de Vida/imunologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Sítios de Ligação/genética , Cobre/metabolismo , DNA Complementar/genética , Levodopa/metabolismo , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNARESUMO
Leishmaniasis is caused by parasitic protozoa transmitted by the bite of a female sand fly and is currently endemic in 88 countries. BALB/c mice are highly susceptible to the infection with the parasite Leishmania major, and this susceptibility has been attributed, in part, to the expansion of Th2 cells, production of their cytokines, and downregulation of Th1 cytokine, interferon gamma (IFN-γ). In this report, we used both aqueous and alcoholic extracts of Iranian borage (Echium amoenum Fisch & C.A. Mey) for treatment of L. major infection in BALB/c mice. We found that both extracts had immunomodulatory properties and increased the level of IFN-γ and lowered the parasite burden in the proximal lymph nodes and prevented the necrosis of the footpad as compared with the untreated infected mice. These results may provide a basis for further studies directed toward the use of the Iranian borage against L. major infection.
Assuntos
Echium/química , Fatores Imunológicos/farmacologia , Leishmania major/efeitos dos fármacos , Leishmaniose/tratamento farmacológico , Estágios do Ciclo de Vida/efeitos dos fármacos , Linfonodos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Animais , Proliferação de Células/efeitos dos fármacos , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Feminino , Flores/química , Fatores Imunológicos/química , Injeções Intraperitoneais , Interferon gama/análise , Interferon gama/biossíntese , Leishmania major/imunologia , Leishmaniose/imunologia , Leishmaniose/parasitologia , Estágios do Ciclo de Vida/imunologia , Linfonodos/imunologia , Linfonodos/parasitologia , Camundongos , Camundongos Endogâmicos BALB C , Extratos Vegetais/química , Células Th1/efeitos dos fármacos , Células Th1/imunologia , Células Th2/efeitos dos fármacos , Células Th2/imunologiaRESUMO
Among the three clinical forms (cutaneous, mucosal and visceral) of leishmaniasis visceral (VL) one is the most devastating type caused by the invasion of the reticuloendothelial system of human by Leishmania donovani, L. infantum and L. chagasi. India and Sudan account for about half the world's burden of VL. Current control strategy is based on chemotherapy, which is difficult to administer, expensive and becoming ineffective due to the emergence of drug resistance. An understanding of resistance mechanism(s) operating in clinical isolates might provide additional leads for the development of new drugs. Further, due to the lack of fully effective treatment the search for novel immune targets is also needed. So far, no vaccine exists for VL despite indications of naturally developing immunity. Therefore, an urgent need for new and effective leishmanicidal agents and for this identification of novel drug and vaccine targets is imperative. The availability of the complete genome sequence of Leishmania has revolutionised many areas of leishmanial research and facilitated functional genomic studies as well as provided a wide range of novel targets for drug designing. Most notably, proteomics and transcriptomics have become important tools in gaining increased understanding of the biology of Leishmania to be explored on a global scale, thus accelerating the pace of discovery of vaccine/drug targets. In addition, these approaches provide the information regarding genes and proteins that are expressed and under which conditions. This review provides a comprehensive view about those proteins/genes identified using proteomics and transcriptomic tools for the development of vaccine/drug against VL.
Assuntos
Perfilação da Expressão Gênica/métodos , Vacinas contra Leishmaniose/imunologia , Leishmaniose Visceral/imunologia , Proteômica/métodos , Animais , Avaliação Pré-Clínica de Medicamentos/métodos , Humanos , Leishmania/efeitos dos fármacos , Leishmania/fisiologia , Vacinas contra Leishmaniose/genética , Vacinas contra Leishmaniose/metabolismo , Leishmaniose Visceral/tratamento farmacológico , Leishmaniose Visceral/prevenção & controle , Estágios do Ciclo de Vida/efeitos dos fármacos , Estágios do Ciclo de Vida/genética , Estágios do Ciclo de Vida/imunologia , Tripanossomicidas/farmacologia , Tripanossomicidas/uso terapêuticoRESUMO
Se analiza la evidencia científica disponible sobre las recomendaciones nutricionales en las distintas etapas de la vida a través de 10 preguntas clave que se cuestionan alguno de los hechos más relevantes en cuanto a las modificaciones dietéticas a lo largo del ciclo vital: ¿cuál debe ser la duración de la lactancia materna?, ¿cuáles son los mejores métodos para promocionar la lactancia materna?, ¿deben utilizarse fórmulas con proteínas de soja para prevenir enfermedades alérgicas en lactantes de alto riesgo?, ¿está indicada la suplementación con micronutrientes en el lactante?, ¿cómo prevenir la obesidad infantil?, ¿existen estrategias válidas de prevención de los trastornos alimentarios en adolescentes?, ¿qué alimentación debemos recomendar para la embarazada y la mujer lactante?, ¿deben utilizarse suplementos nutricionales en los ancianos con riesgo de desnutrición?, ¿deben utilizarse suplementos de micronutrientes en los ancianos para prevenir o tratar las enfermedades degenerativas? (AU)
We review the scientific evidence available on nutritional recommendations in the distinct stages of life through 10 key questions relating to dietary modifications throughout the life cycle, namely: how long should breast-feeding last? What are the best methods for promoting breast-feeding? Should soya infant formulas be used to prevent allergic diseases in infants at high risk? Is micronutrient supplementation indicated in infants? How should childhood obesity be prevented? Are there any valid strategies for preventing eating disorders in adolescents? Which type of diet should be recommended in pregnant and breast-feeding women? Should nutritional supplements be used in elderly individuals at risk of malnutrition? Should micronutrient supplements be used in the elderly to prevent or treat degenerative diseases? (AU)