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1.
Fish Shellfish Immunol ; 56: 303-309, 2016 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-27452973

RESUMEN

Hepcidin, an antimicrobial peptide, plays a crucial role in innate immune system of teleost fish. As a cysteine-rich peptide, hepcidin possesses a dual function including iron regulation and innate immunity. In the present study, a full-length hepcidin cDNA (HtHep) was cloned and characterized by RT-PCR and RACE techniques from taimen (Hucho taimen, Pallas), which is a type of rare, precious and cold-water fish species in China. The cDNA contains an open reading frame (ORF) of 267 bp encoding 88 amino acid (aa), with 170 bp located in the 5(') untranslated region (UTR) and 151 bp in the 3' UTR. The genomic sequences analysis showed that the HtHep gene consisted of three exons and two introns (with the length 94 and 251 bp, respectively). With a predicted molecular mass of 2881.4 Da and a theoretical pI of 8.53, the deduced amino acid encodes a signal peptide of 24 aa, prodomain of 39 aa and mature peptide of 25 aa. The signal peptidase (SA-VP) and the motif RX (K/R)R of propeptide convertase suggested the cleavage site of signal and mature peptide. Eight conserved cysteine residues were also identified and formed four disulfide bonds. Pair-wise alignments showed that HtHep clustered together with two fish species of Salmonidae family (Salmo salar and Oncorhynchus mykiss) in HAMP1 branch. Quantitative RT-PCR analysis indicated that the mRNA levels of HtHep were detected in a wide range of tissues and the highest level was detected in the liver. Its expression was also detected early during embryonic stage and could be up-regulated in the liver when challenged with pathogenic bacteria (Yersinia ruckeri). The recombinant HtHep (rHtHep) had antimicrobial activity against both gram-positive (Micrococcus lysodeikticus and Staphylococcus aureus) and gram-negative bacteria (Escherichia coli). Our results suggested that HtHep might be involved in the innate immune defense against bacterial pathogens in taimen.


Asunto(s)
Enfermedades de los Peces/genética , Proteínas de Peces/genética , Hepcidinas/genética , Yersiniosis/veterinaria , Yersinia ruckeri/fisiología , Secuencia de Aminoácidos , Animales , Clonación Molecular , ADN Complementario/genética , ADN Complementario/metabolismo , Escherichia coli/inmunología , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/microbiología , Proteínas de Peces/química , Proteínas de Peces/metabolismo , Bacterias Grampositivas/inmunología , Hepcidinas/química , Hepcidinas/metabolismo , Filogenia , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Salmonidae , Alineación de Secuencia/veterinaria , Yersiniosis/genética , Yersiniosis/inmunología , Yersiniosis/microbiología
2.
J Immunol Res ; 2015: 482863, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26783541

RESUMEN

Intracellular killing of bacteria is one of the fundamental mechanisms against invading pathogens. Impaired intracellular killing of bacteria by phagocytes may be the reason of chronic infections and may be caused by antibiotics or substances that can be produced by some bacteria. Therefore, it was of great practical importance to examine whether phage preparations may influence the process of phagocyte intracellular killing of bacteria. It may be important especially in the case of patients qualified for experimental phage therapy (approximately half of the patients with chronic bacterial infections have their immunity impaired). Our analysis included 51 patients with chronic Gram-negative and Gram-positive bacterial infections treated with phage preparations at the Phage Therapy Unit in Wroclaw. The aim of the study was to investigate the effect of experimental phage therapy on intracellular killing of bacteria by patients' peripheral blood monocytes and polymorphonuclear neutrophils. We observed that phage therapy does not reduce patients' phagocytes' ability to kill bacteria, and it does not affect the activity of phagocytes in patients with initially reduced ability to kill bacteria intracellularly. Our results suggest that experimental phage therapy has no significant adverse effects on the bactericidal properties of phagocytes, which confirms the safety of the therapy.


Asunto(s)
Bacteriófagos/química , Terapia Biológica/métodos , Infecciones por Bacterias Gramnegativas/terapia , Infecciones por Bacterias Grampositivas/terapia , Monocitos/inmunología , Neutrófilos/inmunología , Bacteriófagos/fisiología , Estudios de Casos y Controles , Bacterias Gramnegativas/inmunología , Bacterias Gramnegativas/virología , Infecciones por Bacterias Gramnegativas/inmunología , Infecciones por Bacterias Gramnegativas/microbiología , Infecciones por Bacterias Gramnegativas/patología , Bacterias Grampositivas/inmunología , Bacterias Grampositivas/virología , Infecciones por Bacterias Grampositivas/inmunología , Infecciones por Bacterias Grampositivas/microbiología , Infecciones por Bacterias Grampositivas/patología , Humanos , Monocitos/microbiología , Neutrófilos/microbiología , Seguridad del Paciente , Fagocitosis/inmunología , Cultivo Primario de Células , Resultado del Tratamiento
3.
Amino Acids ; 46(10): 2403-13, 2014 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-25023447

RESUMEN

This study was conducted to determine effects of dietary supplementation with 1 % L-glutamine for 14 days on the abundance of intestinal bacteria and the activation of intestinal innate immunity in mice. The measured variables included (1) the abundance of Bacteroidetes, Firmicutes, Lactobacillus, Streptococcus and Bifidobacterium in the lumen of the small intestine; (2) the expression of toll-like receptors (TLRs), pro-inflammatory cytokines, and antibacterial substances secreted by Paneth cells and goblet cells in the jejunum, ileum and colon; and (3) the activation of TLR4-nuclear factor kappa B (NF-κB), mitogen-activated protein kinases (MAPK), and phosphoinositide-3-kinases (PI3K)/PI3K-protein kinase B (Akt) signaling pathways in the jejunum and ileum. In the jejunum, glutamine supplementation decreased the abundance of Firmicutes, while increased mRNA levels for antibacterial substances in association with the activation of NF-κB and PI3K-Akt pathways. In the ileum, glutamine supplementation induced a shift in the Firmicutes:Bacteroidetes ratio in favor of Bacteroidetes, and enhanced mRNA levels for Tlr4, pro-inflammatory cytokines, and antibacterial substances participating in NF-κB and JNK signaling pathways. These results indicate that the effects of glutamine on the intestine vary with its segments and compartments. Collectively, dietary glutamine supplementation of mice beneficially alters intestinal bacterial community and activates the innate immunity in the small intestine through NF-κB, MAPK and PI3K-Akt signaling pathways.


Asunto(s)
Suplementos Dietéticos , Glutamina/administración & dosificación , Inmunidad Innata , Factores Inmunológicos/administración & dosificación , Mucosa Intestinal/microbiología , Intestino Delgado/microbiología , Animales , Bacteroidetes/clasificación , Bacteroidetes/crecimiento & desarrollo , Bacteroidetes/inmunología , Bacteroidetes/aislamiento & purificación , Colon/inmunología , Colon/metabolismo , Citocinas/genética , Citocinas/metabolismo , Femenino , Células Caliciformes/inmunología , Células Caliciformes/metabolismo , Bacterias Grampositivas/clasificación , Bacterias Grampositivas/crecimiento & desarrollo , Bacterias Grampositivas/inmunología , Bacterias Grampositivas/aislamiento & purificación , Íleon/citología , Íleon/inmunología , Íleon/metabolismo , Íleon/microbiología , Mucosa Intestinal/citología , Mucosa Intestinal/inmunología , Mucosa Intestinal/metabolismo , Intestino Delgado/citología , Intestino Delgado/inmunología , Intestino Delgado/metabolismo , Yeyuno/citología , Yeyuno/inmunología , Yeyuno/metabolismo , Yeyuno/microbiología , Ratones Endogámicos ICR , Tipificación Molecular , Células de Paneth/inmunología , Células de Paneth/metabolismo , Distribución Aleatoria , Transducción de Señal , Organismos Libres de Patógenos Específicos , Receptor Toll-Like 4/genética , Receptor Toll-Like 4/metabolismo
4.
Br J Nutr ; 112(1): 15-29, 2014 Jul 14.
Artículo en Inglés | MEDLINE | ID: mdl-24774835

RESUMEN

The aim of the present study was to investigate the effects of different dietary sustained-release microencapsulated sodium butyrate (MSB) products (0 (non-supplement), 1·5 and 3·0 h) for a control or oxidised soyabean oil (SBO) diet on fish production, intestinal mucosal condition, immunity and intestinal bacteria in juvenile common carp (Cyprinus carpio). Dietary MSB increased weight gain and reduced the feed conversion ratio within the control and oxidised SBO groups. Gut mucosa was damaged in the oxidised SBO group fed without MSB, in contrast to a normal appearance found in fish fed the MSB1·5 and MSB3·0 diets in the oxidised SBO group. Microvillus density increased in fish fed the MSB1·5 and MSB3·0 diets in the oxidised SBO group (P< 0·001); however, microvillus density was affected by the different pre-fed diets in the midgut (P< 0·001) and by the different sustained-release times of MSB in the distal gut (DG) (P= 0·003). The interaction between the pre-fed diets and the sustained-release times of dietary MSB was significant for the relative gene expression levels of gut heat shock protein-70 (HSP70), pro-inflammatory cytokines (IL-1ß and TNF-α) and anti-inflammatory cytokines (transforming growth factor-ß) within each gut segment, except for HSP70 in the DG and IL-1ß in the foregut. Modulation of adherent bacterial communities within each gut segment investigated was not obvious when the common carp were fed the diets with MSB, as similarity coefficients of >0·79 were observed. These results indicated that MSB can be used as a dietary supplement to repair or prevent intestinal damage in carp fed oxidised SBO.


Asunto(s)
Antioxidantes/metabolismo , Adhesión Bacteriana , Ácido Butírico/metabolismo , Carpas/crecimiento & desarrollo , Dieta/veterinaria , Inmunidad Mucosa , Mucosa Intestinal/crecimiento & desarrollo , Animales , Antioxidantes/química , Acuicultura , Ácido Butírico/química , Carpas/inmunología , Carpas/metabolismo , Carpas/microbiología , China , Citocinas/genética , Citocinas/metabolismo , Proteínas de Peces/genética , Proteínas de Peces/metabolismo , Regulación del Desarrollo de la Expresión Génica , Bacterias Gramnegativas/crecimiento & desarrollo , Bacterias Gramnegativas/inmunología , Bacterias Gramnegativas/aislamiento & purificación , Bacterias Grampositivas/crecimiento & desarrollo , Bacterias Grampositivas/inmunología , Bacterias Grampositivas/aislamiento & purificación , Proteínas HSP70 de Choque Térmico/genética , Proteínas HSP70 de Choque Térmico/metabolismo , Mucosa Intestinal/inmunología , Mucosa Intestinal/microbiología , Mucosa Intestinal/ultraestructura , Intestinos/crecimiento & desarrollo , Intestinos/inmunología , Intestinos/microbiología , Intestinos/ultraestructura , Microvellosidades/inmunología , Microvellosidades/microbiología , Microvellosidades/ultraestructura , Oxidación-Reducción , Estrés Oxidativo , Solubilidad , Aceite de Soja/efectos adversos , Aceite de Soja/antagonistas & inhibidores , Aceite de Soja/química
5.
Br J Nutr ; 111(3): 387-402, 2014 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-23931069

RESUMEN

The human intestine is colonised by 10¹³ to 10¹4 micro-organisms, the vast majority of which belong to the phyla Firmicutes and Bacteroidetes. Although highly stable over time, the composition and activities of the microbiota may be influenced by a number of factors including age, diet and antibiotic treatment. Although perturbations in the composition or functions of the microbiota are linked to inflammatory and metabolic disorders (e.g. inflammatory bowel diseases, irritable bowel syndrome and obesity), it is unclear at this point whether these changes are a symptom of the disease or a contributing factor. A better knowledge of the mechanisms through which changes in microbiota composition (dysbiosis) promote disease states is needed to improve our understanding of the causal relationship between the gut microbiota and disease. While evidence of the preventive and therapeutic effects of probiotic strains on diarrhoeal illness and other intestinal conditions is promising, the exact mechanisms of the beneficial effects are not fully understood. Recent studies have raised the question of whether non-viable probiotic strains can confer health benefits on the host by influencing the immune system. As the potential health effect of these non-viable bacteria depends on whether the mechanism of this effect is dependent on viability, future research needs to consider each probiotic strain on a case-by-case basis. The present review provides a comprehensive, updated overview of the human gut microbiota, the factors influencing its composition and the role of probiotics as a therapeutic modality in the treatment and prevention of diseases and/or restoration of human health.


Asunto(s)
Envejecimiento , Dieta , Estado de Salud , Intestinos/microbiología , Modelos Biológicos , Probióticos/uso terapéutico , Animales , Antibacterianos/efectos adversos , Dieta/efectos adversos , Suplementos Dietéticos , Bacterias Gramnegativas/efectos de los fármacos , Bacterias Gramnegativas/crecimiento & desarrollo , Bacterias Gramnegativas/inmunología , Bacterias Grampositivas/efectos de los fármacos , Bacterias Grampositivas/crecimiento & desarrollo , Bacterias Grampositivas/inmunología , Humanos , Enfermedades Inflamatorias del Intestino/dietoterapia , Enfermedades Inflamatorias del Intestino/inmunología , Enfermedades Inflamatorias del Intestino/microbiología , Enfermedades Inflamatorias del Intestino/prevención & control , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/crecimiento & desarrollo , Mucosa Intestinal/inmunología , Mucosa Intestinal/microbiología , Intestinos/efectos de los fármacos , Intestinos/crecimiento & desarrollo , Intestinos/inmunología , Síndrome del Colon Irritable/dietoterapia , Síndrome del Colon Irritable/inmunología , Síndrome del Colon Irritable/microbiología , Síndrome del Colon Irritable/prevención & control , Viabilidad Microbiana/efectos de los fármacos
6.
J. pediatr. (Rio J.) ; 89(4): 394-399, ju.-ago. 2013. ilus
Artículo en Portugués | LILACS | ID: lil-684139

RESUMEN

OBJETIVO: Comparar o crescimento bacteriano em colostro puro e colostro com aditivo do leite materno contendo ferro. MÉTODOS: Foram comparadas 78 amostras de colostro puro ou colostro com adição de aditivo do leite materno contendo ferro para avaliar o crescimento de Escherichia coli, Staphylococcus aureus e Pseudomonas aeruginosa. Para a análise qualitativa, discos de papel-filtro foram imersos em amostras de cada grupo e incubados por 48 horas com 10¹ Unidades Formadoras de Colônias/mL de cada cepa. Para a avaliação quantitativa, 1 mL de cada cepa contendo 10(7) Unidades Formadoras de Colônias/mL foi homogeneizado com 1 mL, tanto de colostro puro quanto de colostro com aditivo do leite materno, espalhado em placa de Petri e incubado a 37ºC. O número de Unidades Formadoras de Colônias foi contado 24 horas depois. RESULTADOS: A análise qualitativa não mostrou nenhuma diferença no crescimento bacteriano. Na avaliação quantitativa, o crescimento de Escherichia coli (EC) no grupo C foi de 29,4±9,7 x 10(6) CFU/mL, enquanto no grupo FM85 foi de 31,2±10,8 x 10(6) CFU/mL. A diferença entre o crescimento médio foi de 1,9±4,9 x 10(6) CFU/mL (p = 0,001). Não houve diferenças no crescimento de Staphylococcus aureus e Pseudomonas aeruginosa. CONCLUSÃO: A adição de ferro a essa concentração reduz a ação bacteriostática do leite materno contra Escherichia coli.


OBJECTIVE: To compare bacterial growth in pure colostrum versus colostrum with human milk fortifier (HMF) containing iron. METHODS: The growth of Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa in 78 samples of pure colostrum or colostrum with added iron-containing HMF was compared. For qualitative analysis, filter paper discs were immersed in samples from each group and incubated for 48 hours with 10¹ colony forming units (CFUs)/mL of each strain. For quantitative assessment, 1 mL of each strain containing 10(7) CFUs/mL was homogenized with 1 mL of either colostrum or colostrum with human milk fortifier, seeded into a Petri dish, and incubated at 37ºC. Twenty-four hours later, the number of CFUs was counted. RESULTS: The qualitative analysis showed no difference in bacterial growth. In the quantitative evaluation, E. coli growth in the control group was 29.4±9.7 x 10(6) CFU/ mL, while in the HMF group it was 31.2±10.8 x 10(6) CFU/mL. The difference between the average growth was 1.9±4.9 x 10(6) CFU/mL (p = 0.001). There were no differences in S. aureus and P. aeruginosa growth. CONCLUSION: Addition of iron at this concentration reduces breast milk bacteriostatic action against E. coli.


Asunto(s)
Animales , Femenino , Humanos , Embarazo , Calostro/microbiología , Alimentos Fortificados , Bacterias Gramnegativas/crecimiento & desarrollo , Bacterias Grampositivas/crecimiento & desarrollo , Infecciones por Bacterias Grampositivas/inmunología , Hierro , Leche Humana , Calostro/inmunología , Escherichia coli/crecimiento & desarrollo , Bacterias Gramnegativas/inmunología , Bacterias Grampositivas/inmunología , Infecciones por Bacterias Grampositivas/prevención & control , Hierro/administración & dosificación , Lactoferrina/fisiología , Pseudomonas aeruginosa/crecimiento & desarrollo , Staphylococcus aureus/crecimiento & desarrollo
7.
J Pediatr (Rio J) ; 89(4): 394-9, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-23791235

RESUMEN

OBJECTIVE: To compare bacterial growth in pure colostrum versus colostrum with human milk fortifier (HMF) containing iron. METHODS: The growth of Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa in 78 samples of pure colostrum or colostrum with added iron-containing HMF was compared. For qualitative analysis, filter paper discs were immersed in samples from each group and incubated for 48 hours with 10(1) colony forming units (CFUs)/mL of each strain. For quantitative assessment, 1 mL of each strain containing 10(7) CFUs/mL was homogenized with 1 mL of either colostrum or colostrum with human milk fortifier, seeded into a Petri dish, and incubated at 37°C. Twenty-four hours later, the number of CFUs was counted. RESULTS: The qualitative analysis showed no difference in bacterial growth. In the quantitative evaluation, E. coli growth in the control group was 29.4±9.7×10(6)CFU/mL, while in the HMF group it was 31.2±10.8×10(6)CFU/mL. The difference between the average growth was 1.9±4.9×10(6)CFU/mL (p=0.001). There were no differences in S. aureus and P. aeruginosa growth. CONCLUSION: Addition of iron at this concentration reduces breast milk bacteriostatic action against E. coli.


Asunto(s)
Calostro/microbiología , Alimentos Fortificados , Bacterias Gramnegativas/crecimiento & desarrollo , Bacterias Grampositivas/crecimiento & desarrollo , Infecciones por Bacterias Grampositivas/inmunología , Hierro , Leche Humana , Animales , Calostro/inmunología , Escherichia coli/crecimiento & desarrollo , Femenino , Bacterias Gramnegativas/inmunología , Bacterias Grampositivas/inmunología , Infecciones por Bacterias Grampositivas/prevención & control , Humanos , Hierro/administración & dosificación , Lactoferrina/fisiología , Embarazo , Pseudomonas aeruginosa/crecimiento & desarrollo , Staphylococcus aureus/crecimiento & desarrollo
8.
PLoS One ; 7(5): e36783, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-22611464

RESUMEN

Complement C3 plays an essential role in the opsonization of pathogens in the mammalian complement system, whereas the molecular mechanism underlying C3 activation in invertebrates remains unknown. To understand the molecular mechanism of C3b deposition on microbes, we characterized two types of C2/factor B homologs (designated TtC2/Bf-1 and TtC2/Bf-2) identified from the horseshoe crab Tachypleus tridentatus. Although the domain architectures of TtC2/Bf-1 and TtC2/Bf-2 were identical to those of mammalian homologs, they contained five-repeated and seven-repeated complement control protein domains at their N-terminal regions, respectively. TtC2/Bf-1 and TtC2/Bf-2 were synthesized and glycosylated in hemocytes and secreted to hemolymph plasma, which existed in a complex with C3 (TtC3), and their activation by microbes was absolutely Mg(2+)-dependent. Flow cytometric analysis revealed that TtC3b deposition was Mg(2+)-dependent on Gram-positive bacteria or fungi, but not on Gram-negative bacteria. Moreover, this analysis demonstrated that Ca(2+)-dependent lectins (C-reactive protein-1 and tachylectin-5A) were required for TtC3b deposition on Gram-positive bacteria, and that a Ca(2+)-independent lectin (Tachypleus plasma lectin-1) was definitely indispensable for TtC3b deposition on fungi. In contrast, a horseshoe crab lipopolysaccharide-sensitive protease factor C was necessary and sufficient to deposit TtC3b on Gram-negative bacteria. We conclude that plasma lectins and factor C play key roles in microbe-specific TtC3b deposition in a C2/factor B-dependent or -independent manner.


Asunto(s)
Complemento C2/inmunología , Complemento C3b/inmunología , Factor B del Complemento/inmunología , Cangrejos Herradura/inmunología , Animales , Clonación Molecular , Activación de Complemento/inmunología , Complemento C2/genética , Complemento C2/metabolismo , Complemento C3b/metabolismo , Factor B del Complemento/genética , Factor B del Complemento/metabolismo , ADN Complementario , Factores de Transcripción Forkhead/inmunología , Factores de Transcripción Forkhead/metabolismo , Hongos/inmunología , Bacterias Grampositivas/inmunología , Cangrejos Herradura/microbiología , Magnesio/metabolismo , Modelos Biológicos , Unión Proteica/inmunología , Estructura Terciaria de Proteína
9.
Fish Shellfish Immunol ; 32(5): 716-23, 2012 May.
Artículo en Inglés | MEDLINE | ID: mdl-22342746

RESUMEN

C-type lectins are a family of calcium-dependent carbohydrate-binding proteins. In the present study, a C-type lectin (designated as AiCTL5) was identified and characterized from Argopecten irradians. The full-length cDNA of AiCTL5 was of 673 bp, containing a 5' untranslated region (UTR) of 24 bp, a 3' UTR of 130 bp with a poly (A) tail, and an open reading frame (ORF) of 519 bp encoding a polypeptide of 172 amino acids with a putative signal peptide of 17 amino acids. A C-type lectin-like domain (CRD) containing 6 conserved cysteines and a putative glycosylation sites were identified in the deduced amino acid sequence of AiCTL5. AiCTL5 shared 11%-27.5% identity with the previous reported C-type lectin from A. irradians. The cDNA fragment encoding the mature peptide of AiCTL5 was recombined into pET-21a (+) with a C-terminal hexa-histidine tag fused in-frame, and expressed in Escherichia coli Origami (DE3). The recombinant AiCTL5 (rAiCTL5) agglutinated Gram-negative E. coli TOP10F' and Listonella anguillarum, but did not agglutinate Gram-positive bacteria Bacillus thuringiensis and Micrococcus luteus, and the agglutination could be inhibited by EDTA, indicating that AiCTL5 was a Ca(2+)-dependent lectin. rAiCTL5 exhibited a significantly strong activity to bind LPS from E. coli, which conformed to the agglutinating activity toward Gram-negative bacteria. Moreover, rAiCTL5 also agglutinated rabbit erythrocytes. These results indicated that AiCTL5 could function as a pattern recognition receptor to protect bay scallop from Gram-negative bacterial infection, and also provide evidence to understand the structural and functional diverse of lectin.


Asunto(s)
Lectinas Tipo C/metabolismo , Pectinidae/genética , Pectinidae/inmunología , Aglutinación , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , ADN Complementario/genética , Eritrocitos/inmunología , Perfilación de la Expresión Génica/veterinaria , Bacterias Gramnegativas/genética , Bacterias Gramnegativas/inmunología , Bacterias Grampositivas/genética , Bacterias Grampositivas/inmunología , Inmunidad Innata , Lectinas Tipo C/química , Lectinas Tipo C/genética , Lipopolisacáridos/metabolismo , Lipopolisacáridos/farmacología , Datos de Secuencia Molecular , Pectinidae/microbiología , Filogenia , Estructura Terciaria de Proteína , Conejos , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Alineación de Secuencia/veterinaria
10.
Am J Physiol Endocrinol Metab ; 302(3): E374-86, 2012 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-22094473

RESUMEN

Low-grade inflammation observed in obesity is a risk factor for cardiovascular disease. Recent studies revealed that this would be linked to gut-derived endotoxemia during fat digestion in high-fat diets, but nothing is known about the effect of lipid composition. The study was designed to test the impact of oil composition of high-fat diets on endotoxin metabolism and inflammation in mice. C57/Bl6 mice were fed for 8 wk with chow or isocaloric isolipidic diets enriched with oils differing in fatty acid composition: milk fat, palm oil, rapeseed oil, or sunflower oil. In vitro, adipocytes (3T3-L1) were stimulated or not with lipopolysaccharide (LPS; endotoxin) and incubated with different fatty acids. In mice, the palm group presented the highest level of IL-6 in plasma (P < 0.01) together with the highest expression in adipose tissue of IL-1ß and of LPS-sensing TLR4 and CD14 (P < 0.05). The higher inflammation in the palm group was correlated with a greater ratio of LPS-binding protein (LBP)/sCD14 in plasma (P < 0.05). The rapeseed group resulted in higher sCD14 than the palm group, which was associated with lower inflammation in both plasma and adipose tissue despite higher plasma endotoxemia. Taken together, our results reveal that the palm oil-based diet resulted in the most active transport of LPS toward tissues via high LBP and low sCD14 and the greatest inflammatory outcomes. In contrast, a rapeseed oil-based diet seemed to result in an endotoxin metabolism driven toward less inflammatory pathways. This shows that dietary fat composition can contribute to modulate the onset of low-grade inflammation through the quality of endotoxin receptors.


Asunto(s)
Tejido Adiposo Blanco/inmunología , Citocinas/metabolismo , Dieta Alta en Grasa/efectos adversos , Enfermedades Metabólicas/etiología , Enfermedades Metabólicas/inmunología , Receptores Inmunológicos/metabolismo , Células 3T3-L1 , Proteínas de Fase Aguda , Tejido Adiposo Blanco/metabolismo , Animales , Biomarcadores/sangre , Biomarcadores/metabolismo , Proteínas Portadoras/sangre , Citocinas/sangre , Ácidos Grasos Monoinsaturados , Ácidos Grasos no Esterificados/efectos adversos , Ácidos Grasos no Esterificados/sangre , Bacterias Gramnegativas/inmunología , Bacterias Gramnegativas/aislamiento & purificación , Bacterias Grampositivas/inmunología , Bacterias Grampositivas/aislamiento & purificación , Intestinos/inmunología , Intestinos/microbiología , Intestinos/patología , Receptores de Lipopolisacáridos/sangre , Receptores de Lipopolisacáridos/metabolismo , Masculino , Glicoproteínas de Membrana/sangre , Enfermedades Metabólicas/metabolismo , Enfermedades Metabólicas/microbiología , Ratones , Ratones Endogámicos C57BL , Aceite de Palma , Aceites de Plantas/efectos adversos , Distribución Aleatoria , Aceite de Brassica napus , Aceite de Girasol , Receptor Toll-Like 4/metabolismo
11.
Clin Exp Allergy ; 42(1): 76-84, 2012 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-22092824

RESUMEN

BACKGROUND: Recently, it has been established that pollen grains contain Th2-enhancing activities besides allergens. OBJECTIVE: The aim of this study was to analyse whether pollen carry additional adjuvant factors like microbes and what immunological effects they may exert. METHODS: Timothy pollen grains were collected and disseminated on agar plates, and the growing microorganisms were cultivated and defined. Furthermore, the immunologic effects of microbial products on DC and T cell responses were analysed. RESULTS: A complex mixture of bacteria and moulds was detected on grass pollen. Besides Gram-negative bacteria that are known to favour Th1-directed immune responses, moulds were identified as being sources of allergens themselves. Herein, we focused on Gram-positive bacteria that were found in high numbers, e.g. Bacillus cereus and Bacillus subtilis. Contact of immature dendritic cells (DC) from grass pollen allergic donors with supernatants of homogenized Gram-positive bacteria induced maturation of DC as measured by up-regulation of CD80, CD83 and CD86 and by enhanced production of IL-6, IL-12p40 and TNF-α, which was less pronounced compared with effects induced by lipopolysaccharide (LPS). Consequently, stimulation of autologous CD4(+) T cells with supernatants of homogenized Gram-positive bacteria plus grass pollen allergen-pulsed DC led to an enhanced proliferation and production of IL-4, IL-13, IL-10, IL-17, IL-22 and IFN-γ production compared with T cells that were stimulated with allergen-pulsed immature DC alone, whereas production of the transcription factor for regulatory T cells FoxP3 was not significantly affected. CONCLUSIONS AND CLINICAL RELEVANCE: These data indicate that grass pollen is colonized by several microorganisms that influence the immune response differently. Similar to LPS, supernatants of homogenized Gram-positive bacteria may serve as adjuvants by augmenting DC maturation and inflammatory Th1, Th2 and Th17 responses helping to initiate allergic immune responses.


Asunto(s)
Bacterias Grampositivas/inmunología , Inflamación/inmunología , Activación de Linfocitos/inmunología , Phleum/microbiología , Polen/microbiología , Rinitis Alérgica Estacional/inmunología , Adyuvantes Inmunológicos , Bacillus cereus/inmunología , Bacillus cereus/aislamiento & purificación , Bacillus subtilis/inmunología , Bacillus subtilis/aislamiento & purificación , Diferenciación Celular , Medios de Cultivo , Células Dendríticas/citología , Células Dendríticas/inmunología , Citometría de Flujo , Bacterias Grampositivas/clasificación , Bacterias Grampositivas/aislamiento & purificación , Humanos , Phleum/inmunología , Polen/inmunología , Rinitis Alérgica Estacional/fisiopatología , Células TH1/inmunología , Células Th17/inmunología , Células Th2/inmunología
12.
J Immunol ; 181(2): 1083-95, 2008 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-18606660

RESUMEN

Following trauma, the CNS of the medicinal leech, unlike the mammalian CNS, has a strong capacity to regenerate neurites and synaptic connections that restore normal function. In this study, we show that this regenerative process is enhanced by a controlled bacterial infection, suggesting that induction of regeneration of normal CNS function may depend critically upon the coinitiation of an immune response. We explore the interaction between the activation of a neuroimmune response and the process of regeneration by assaying the potential roles of two newly characterized antimicrobial peptides. Our data provide evidence that microbial components differentially induce the transcription, by microglial cells, of both antimicrobial peptide genes, the products of which accumulate rapidly at sites in the CNS undergoing regeneration following axotomy. Using a preparation of leech CNS depleted of microglial cells, we also demonstrate the production of antimicrobial peptides by neurons. Interestingly, in addition to exerting antibacterial properties, both peptides act as promoters of the regenerative process of axotomized leech CNS. These data are the first to report the neuronal synthesis of antimicrobial peptides and their participation in the immune response and the regeneration of the CNS. Thus, the leech CNS appears as an excellent model for studying the implication of immune molecules in neural repair.


Asunto(s)
Aeromonas/inmunología , Péptidos Catiónicos Antimicrobianos/biosíntesis , Bacterias Grampositivas/inmunología , Hirudo medicinalis/fisiología , Microglía/metabolismo , Neuronas/metabolismo , Secuencia de Aminoácidos , Animales , Antígenos Bacterianos/inmunología , Antígenos Bacterianos/metabolismo , Péptidos Catiónicos Antimicrobianos/química , Péptidos Catiónicos Antimicrobianos/genética , Péptidos Catiónicos Antimicrobianos/inmunología , Axotomía , Secuencia de Bases , Sistema Nervioso Central/citología , Sistema Nervioso Central/inmunología , Sistema Nervioso Central/metabolismo , Sistema Nervioso Central/fisiología , Exocitosis , Hirudo medicinalis/genética , Hirudo medicinalis/inmunología , Hirudo medicinalis/microbiología , Microglía/citología , Microglía/inmunología , Datos de Secuencia Molecular , Regeneración Nerviosa , Neuronas/citología , Neuronas/inmunología , Alineación de Secuencia
13.
Inflammation ; 27(6): 329-32, 2003 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-14760940

RESUMEN

Gram-positive and Gram-negative bacteria induce different cytokine patterns in human mononuclear cells. We have seen that Gram-positives preferentially induce IL-12 and TNF-alpha, whereas Gram-negatives induce more IL-10, IL-6, and IL-8. In this study, we compared the capacity of these two groups of bacteria to induce PGE2. Monocytes stimulated with Gram-negative bacterial species induced much more PGE2 than did Gram-positive bacteria (5600 +/- 330 vs. 1700 +/- 670 pg/mL, p < 0.001). Blocking of COX-2 by NS398 abolished PGE2 production, but did not alter the cytokine patterns induced by Gram-positive and Gram-negative bacteria. We suggest that Gram-positive and Gram-negative bacteria may stimulate different innate effector functions; Gram-positive bacteria promoting cell-mediated effector functions whereas Gram-negative bacteria inducing mediators inhibiting the same.


Asunto(s)
Dinoprostona/biosíntesis , Bacterias Gramnegativas/inmunología , Bacterias Grampositivas/inmunología , Monocitos/microbiología , Adyuvantes Inmunológicos/biosíntesis , Células Cultivadas , Ciclooxigenasa 2 , Citocinas/biosíntesis , Humanos , Inmunidad Celular , Inflamación/microbiología , Isoenzimas/fisiología , Proteínas de la Membrana , Monocitos/inmunología , Prostaglandina-Endoperóxido Sintasas/fisiología
14.
Int Immunol ; 9(12): 1867-74, 1997 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-9466314

RESUMEN

Modulation of allergic immune responses by using adequate adjuvants is a promising concept for future immunotherapy of type I hypersensitivity. In the present study, recombinant Bet v 1 (rBet v 1, the major birch pollen allergen) was conjugated to cross-linked crystalline surface layer proteins (SL) derived from Gram-positive eubacteria. T cell lines (TCL) and clones (TCC) were established from peripheral blood of birch pollen-allergic patients. TCL and TCC were induced either using rBet v 1 alone or rBet v 1/SL conjugates (rBet v 1/SL) as initial antigen stimulus. Cytokine production after re-stimulation with rBet v 1 was investigated. TCL initiated with rBet v 1/SL showed significantly increased IFN-gamma production as compared to rBet v 1 -selected TCL. TCC were established from TCL of five patients. As expected, the majority of CD4+ TCC induced by rBet v 1 (55%) displayed a Th2-like pattern of cytokine production. However, only 21% of Bet v 1-specific TCC isolated from TCL established with the Bet v 1/SL revealed this phenotype. The majority of SL-specific TCC (80%) belonged to the Th1 phenotype. In cultures of peripheral blood mononuclear cells, both, SL and Bet v 1/SL (but not rBet v 1) stimulated the production of high levels of IL-12, a pivotal mediator of Th1 responses. Moreover, stimulation of rBet v 1-induced TCC with rBet v 1/SL led to an increased IFN-gamma production. This effect could be reversed by neutralizing anti-IL-12 mAb. Together these results indicate an adjuvant effect of SL mediated by IL-12. Our results indicate that bacterial components, such as SL, displaying adjuvant effects may be suitable for immunotherapeutical vaccines for type I allergy.


Asunto(s)
Alérgenos/inmunología , Proteínas Bacterianas/inmunología , Epítopos de Linfocito T/inmunología , Interleucina-12/biosíntesis , Proteínas de la Membrana/inmunología , Proteínas de Plantas/inmunología , Polen/inmunología , Linfocitos T/inmunología , Adyuvantes Inmunológicos/farmacología , Antígenos de Plantas , Proteínas Bacterianas/administración & dosificación , Proteínas Bacterianas/farmacología , Citocinas/biosíntesis , Mapeo Epitopo , Bacterias Grampositivas/inmunología , Bacterias Grampositivas/metabolismo , Humanos , Interferón gamma/biosíntesis , Activación de Linfocitos/inmunología , Proteínas de la Membrana/administración & dosificación , Proteínas de la Membrana/farmacología , Fenotipo , Proteínas de Plantas/administración & dosificación , Proteínas de Plantas/farmacología , Subgrupos de Linfocitos T , Linfocitos T/metabolismo
15.
Immunotechnology ; 2(2): 103-13, 1996 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-9373319

RESUMEN

BACKGROUND: Crystalline cell surface layers (S-layers) from Gram-positive eubacteria had been demonstrated as carrier/adjuvants for chemically synthesized tumor-associated oligosaccharide haptens and capsular polysaccharide antigens of Streptococcus pneumoniae strains. OBJECTIVES: The applicability of S-layers as vaccine carrier for treatment of Type I allergy was investigated. STUDY DESIGN: Native or cross-linked S-layer self-assembly products and cell wall preparations from Bacillus sphaericus CCM 2177 and Thermoanaerobacter thermohydrosulfuricus L111-69 and L110-69 were used for immobilization of recombinant major birch pollen allergen Bet v 1. RESULTS AND CONCLUSIONS: Depending on the carrier used, amounts of approximately 20-40 micrograms allergen per mg conjugate could be immobilized. By application of L-glutamic acid dimethyl ester as a spacer, this value could be increased approximately 10-fold. The functionality of the rBet v 1-conjugates was assessed in immunological systems. (i) The presence of intact B-cell epitopes was demonstrated in inhibition experiments using human Bet v 1-specific IgE. (ii) The rBet v 1-S-layer conjugates were immunogenic in mice. (iii) The proliferation of rBet v 1-specific T-cell clones suggested that the peptides created by processing of immobilized Bet v 1 were similar to those derived from natural allergen. (iv) Stimulation of human allergen-specific TH2 lymphocytes with S-layer-conjugated Bet v 1 led to a modulation of the cytokine production pattern from TH2 to TH0/TH1. This study indicates that S-layers may be suitable carriers for few immunotherapeutical vaccines for Type 1 hypersensitivity.


Asunto(s)
Alérgenos/inmunología , Bacillus/inmunología , Proteínas Bacterianas/inmunología , Proteínas de Plantas/inmunología , Polen/inmunología , Adyuvantes Inmunológicos , Alérgenos/química , Animales , Antígenos de Plantas , Bacillus/química , Pared Celular/inmunología , Cristalografía , Citocinas/biosíntesis , Epítopos , Femenino , Bacterias Grampositivas/química , Bacterias Grampositivas/inmunología , Humanos , Hipersensibilidad/terapia , Inmunoglobulina E/inmunología , Inmunoterapia , Ratones , Ratones Endogámicos BALB C , Microscopía Electrónica , Linfocitos T/inmunología , Células Th2/inmunología , Árboles/inmunología , Vacunas Sintéticas/química
16.
Rev. farm. bioquim ; 6(n.único): 23-31, 1985. ilus
Artículo en Portugués | LILACS | ID: lil-139420

RESUMEN

Cerca de treze xantonas isoladas de duas espécies do gênero Haploclathra foram submetidos à açäo de microorganismos-teste. Foram eleitos um fungo (Thielaviopsis paradoxa), uma bactéria Gram-positiva (Corynebacterium michiganense pv michiganense) e uma bactéria Gram-negativa (Pseudomonas syringae pv pisi). Nenhuma das xantonas testadas apresentou resultados positivos na inibiçäo do crescimento dos microorganismos em questäo.


Asunto(s)
Antígenos Bacterianos , Antígenos Fúngicos , Antifúngicos/inmunología , Corynebacterium/inmunología , Extractos Vegetales/inmunología , Pseudomonas/inmunología , Bacterias Gramnegativas/inmunología , Bacterias Grampositivas/inmunología
17.
Rev. farm. bioquim ; 6(n.único): 33-40, 1985. tab
Artículo en Portugués | LILACS | ID: lil-139421

RESUMEN

Foram investigadas as atividades antibacteriana e antifúngica dos extratos éter de petróleo e alcaloídico de folhas de Aristolochia gigantea Mart e Zucc, pelo método de difusäo em agar. O extrato éter de petróleo mostrou atividade contra uma bactéria Gram-positiva. O extrato alcaloídico apresentou atividade contra duas bactérias Gram-positivas. Uma Gram-negativa, näo apresentando qualquer atividade antifúngica.


Asunto(s)
Antígenos Bacterianos/inmunología , Antígenos Fúngicos/inmunología , Antifúngicos/inmunología , Aspergillus niger/inmunología , Bacillus subtilis/inmunología , Candida/inmunología , Escherichia coli/inmunología , Extractos Vegetales/inmunología , Pseudomonas aeruginosa/inmunología , Saccharomyces cerevisiae/inmunología , Staphylococcus aureus/inmunología , Bacterias Gramnegativas/inmunología , Bacterias Grampositivas/inmunología
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