RESUMEN
Karyopherins, including alpha and beta types, are transport proteins in the eukaryotic cell that carry cargoes across nuclear pore complexes into or out of the nucleus. In this study, full open reading frames of one beta and three alpha types of karyopherin were cloned from cDNA of the domestic silkworm (Bombyx mori). The one beta and three alpha types' open reading frames were 2661, 1563, 1515, and 1551 base pairs long, respectively, and coded 886, 520, 504, and 516 amino acids, respectively. The alphas all had one importin-beta-binding (IBB) domain, and eight, four, or seven armadillo/beta-catenin-like repeats. The beta had 19 HEAT repeat domains, which constructed one importin-beta-N-terminal domain and one IBB domain. The recombinant proteins were expressed in Escherichia coli cells. The molecular weight of the beta type was approximately 100 kDa, and the alphas weighed approximately 60 kDa. Phylogenic tree construction revealed that the alphas could be classified into three known karyopherin-alpha subfamilies. We detected mRNA of the four karyopherins in normal 3rd day of 5th instar larvae, and in larvae injected with Gram-positive bacteria, Gram-negative bacteria, viruses, and fungi using real-time fluorescence quantitative reverse transcriptase-polymerase chain reaction, and found that the four karyopherins were widely distributed, but their expression levels were related to tissues type, the microbe injected, and the time point.
Asunto(s)
Bombyx/genética , Bombyx/inmunología , Expresión Génica , Inmunidad , Carioferinas/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Bombyx/clasificación , Clonación Molecular , Carioferinas/química , Carioferinas/metabolismo , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Especificidad de Órganos/genética , Filogenia , ARN Mensajero/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Alineación de Secuencia , Análisis de Secuencia de ADNRESUMEN
BACKGROUND: Declining immune function poses an important clinical challenge worldwide and supplementation with natural products that possessing immune enhancing properties is a promising approach for preventing or delaying immune function decline. Cocoons from yellow silkworms are a significant source of lutein, and this unexplored silk extract could be a viable alternative source for dietary lutein. This study assessed immunomodulatory activities of the silk lutein extract. Female BALB/c mice orally received lutein, either as silk or marigold extracts (10 or 20 mg/kg daily), or vehicle only (1% tween 80 in PBS pH 7.4) for 4 weeks. Natural killer (NK) cell activity, specific antibody production, lymphocyte subpopulations, mitogen-induced lymphocyte proliferation, and cytokine production were examined. RESULTS: Silk lutein extract increased NK cell activity, and the effect was dose-related whereas marigold lutein extract was ineffective. Silk lutein extract dose-dependently enhanced antibody production in pre-immunized mice but marigold lutein extract had no effect. Feeding with silk lutein extract increased the populations of CD3+ and CD4 + CD3 + cells. Silk lutein extract also stimulated concanavalin A- and lipopolysaccharide-induced proliferations of T and B lymphocytes, respectively. Moreover, silk lutein extract increased IL-2 and IFN-γ production while the effect of marigold lutein extract was undetectable. CONCLUSIONS: Together, silk lutein extract enhanced both innate and adaptive immune functions. This preparation may prove to be an effective supplement for strengthened immunity.
Asunto(s)
Exoesqueleto/química , Bombyx/inmunología , Factores Inmunológicos/análisis , Luteína/inmunología , Seda/inmunología , Extractos de Tejidos/inmunología , Animales , Anticuerpos Heterófilos/sangre , Linfocitos B/efectos de los fármacos , Bombyx/metabolismo , Proliferación Celular/efectos de los fármacos , Femenino , Citometría de Flujo , Flores/inmunología , Interferón gamma/análisis , Interleucina-10/análisis , Interleucina-2/análisis , Interleucina-4/análisis , Células Asesinas Naturales/efectos de los fármacos , Células Asesinas Naturales/inmunología , Luteína/aislamiento & purificación , Ratones Endogámicos BALB C , Extractos Vegetales/inmunología , Pupa/inmunología , Pupa/metabolismo , Seda/química , Linfocitos T/efectos de los fármacos , Tagetes/inmunología , Extractos de Tejidos/farmacologíaRESUMEN
OBJECTIVE: this study aimed to prepare a silkworm moth (Bombyx mori) antigenic extract and to perform skin prick tests with this extract in patients with allergic respiratory diseases; to evaluate serum specific immunoglobulin E (IgE) to Bombyx mori using ImmunoCAP(r) system and to report the frequency of positivity between the two methods and with clinical data. METHODS: this was a cross-sectional study with 99 children and adolescents diagnosed with asthma and/or allergic rhinitis, who had skin reactivity to at least one of the six aeroallergens tested. Clinical data were evaluated: skin prick tests with Bombyx mori in-house extract, and total and specific IgE analysis using ImmunoCAP(r) were performed. RESULTS: the frequency of Bombyx mori specific IgE was found to be 52.5% and 60% using the skin prick test and ImmunoCAP(r), respectively. An association between a positive skin test for Bombyx mori and the presence of allergic rhinitis, atopic dermatitis, and urticaria was observed, but the same was not true for asthma or allergic conjunctivitis. There was no relation with the severity of asthma or rhinitis symptoms. CONCLUSIONS: a high frequency of sensitization to Bombyx mori was observed in a selected population of patients with respiratory allergic diseases in the city of Curitiba, state of Paraná, Brazil. The extract prepared from the wings of this moth species is effective in demonstrating this sensitivity. .
OBJETIVO: preparar extrato antigênico da mariposa do bicho-da-seda (Bombyx mori) e realizar testes cutâneos com esse extrato em pacientes com doenças respiratórias alérgicas, avaliar IgE sérica específica para Bombyx mori usando o sistema ImmunoCAP(r) e comparar a frequência de positividade entre os dois métodos e com dados clínicos. MÉTODOS: Estudo transversal com 99 crianças e adolescentes com diagnóstico de asma e/ou rinite alérgica, que apresentaram reação cutânea a pelo menos um dos seis aeroalérgenos testados. Os dados clínicos foram avaliados; testes cutâneos com extrato de Bombyx mori e análise de IgE total e específica por ImmunoCAP(r) foram realizados. RESULTADOS: a frequência de IgE específica para Bombyx mori foi de 52,5% e 60%, respectivamente, pelo teste cutâneo e ImmunoCAP(r). Foi observada uma associação entre o teste cutâneo positivo para Bombyx mori e a presença de rinite alérgica, dermatite atópica e urticária, mas o mesmo não ocorreu para a asma ou conjuntivite alérgica. Não houve relação com a gravidade dos sintomas de asma ou rinite. CONCLUSÕES: alta frequência de sensibilização à Bombyx mori foi encontrada em uma população selecionada de pacientes com doenças alérgicas respiratórias na cidade de Curitiba, estado do Paraná, Brasil. O extrato preparado a partir das asas dessa espécie de mariposa é eficaz em demonstrar essa sensibilidade. .
Asunto(s)
Adolescente , Animales , Niño , Femenino , Humanos , Masculino , Alérgenos/inmunología , Asma/inmunología , Bombyx/inmunología , Rinitis Alérgica Perenne/inmunología , Asma/epidemiología , Brasil , Estudios Transversales , Inmunoglobulina E/sangre , Mariposas Nocturnas/inmunología , Rinitis Alérgica , Rinitis Alérgica Perenne/epidemiología , Pruebas Cutáneas/métodosRESUMEN
OBJECTIVE: this study aimed to prepare a silkworm moth (Bombyx mori) antigenic extract and to perform skin prick tests with this extract in patients with allergic respiratory diseases; to evaluate serum specific immunoglobulin E (IgE) to Bombyx mori using ImmunoCAP® system and to report the frequency of positivity between the two methods and with clinical data. METHODS: this was a cross-sectional study with 99 children and adolescents diagnosed with asthma and/or allergic rhinitis, who had skin reactivity to at least one of the six aeroallergens tested. Clinical data were evaluated: skin prick tests with Bombyx mori in-house extract, and total and specific IgE analysis using ImmunoCAP® were performed. RESULTS: the frequency of Bombyx mori specific IgE was found to be 52.5% and 60% using the skin prick test and ImmunoCAP®, respectively. An association between a positive skin test for Bombyx mori and the presence of allergic rhinitis, atopic dermatitis, and urticaria was observed, but the same was not true for asthma or allergic conjunctivitis. There was no relation with the severity of asthma or rhinitis symptoms. CONCLUSIONS: a high frequency of sensitization to Bombyx mori was observed in a selected population of patients with respiratory allergic diseases in the city of Curitiba, state of Paraná, Brazil. The extract prepared from the wings of this moth species is effective in demonstrating this sensitivity.
Asunto(s)
Alérgenos/inmunología , Asma/inmunología , Bombyx/inmunología , Rinitis Alérgica Perenne/inmunología , Adolescente , Animales , Asma/epidemiología , Brasil , Niño , Estudios Transversales , Femenino , Humanos , Inmunoglobulina E/sangre , Masculino , Mariposas Nocturnas/inmunología , Rinitis Alérgica , Rinitis Alérgica Perenne/epidemiología , Pruebas Cutáneas/métodosRESUMEN
BACKGROUND: Declining immune function poses an important clinical challenge worldwide and supplementation with natural products that possessing immune enhancing properties is a promising approach for preventing or delaying immune function decline. Cocoons from yellow silkworms are a significant source of lutein, and this unexplored silk extract could be a viable alternative source for dietary lutein. This study assessed immunomodulatory activities of the silk lutein extract. Female BALB/c mice orally received lutein, either as silk or marigold extracts (10 or 20 mg/kg daily), or vehicle only (1% tween 80 in PBS pH 7.4) for 4 weeks. Natural killer (NK) cell activity, specific antibody production, lymphocyte subpopulations, mitogen-induced lymphocyte proliferation, and cytokine production were examined. RESULTS: Silk lutein extract increased NK cell activity, and the effect was dose-related whereas marigold lutein extract was ineffective. Silk lutein extract dose-dependently enhanced antibody production in pre-immunized mice but marigold lutein extract had no effect. Feeding with silk lutein extract increased the populations of CD3+ and CD4 + CD3 + cells. Silk lutein extract also stimulated concanavalin A- and lipopolysaccharide-induced proliferations of T and B lymphocytes, respectively. Moreover, silk lutein extract increased IL-2 and IFN-γ production while the effect of marigold lutein extract was undetectable. CONCLUSIONS: Together, silk lutein extract enhanced both innate and adaptive immune functions. This preparation may prove to be an effective supplement for strengthened immunity.