RESUMEN
Almost all effective treatments for non-alcoholic fatty liver disease (NAFLD) involve reduction of adiposity, which suggests the metabolic axis between liver and adipose tissue is essential to NAFLD development. Since excessive dietary sugar intake may be an initiating factor for NAFLD, we have characterized the metabolic effects of liquid sucrose intake at concentrations relevant to typical human consumption in mice. We report that sucrose intake induces sexually dimorphic effects in liver, adipose tissue, and the microbiome; differences concordant with steatosis severity. We show that when steatosis is decoupled from impairments in insulin responsiveness, sex is a moderating factor that influences sucrose-driven lipid storage and the contribution of de novo fatty acid synthesis to the overall hepatic triglyceride pool. Our findings provide physiologic insight into how sex influences the regulation of adipose-liver crosstalk and highlight the importance of extrahepatic metabolism in the pathogenesis of diet-induced steatosis and NAFLD.
Asunto(s)
Enfermedad del Hígado Graso no Alcohólico , Tejido Adiposo/metabolismo , Animales , Sacarosa en la Dieta/efectos adversos , Ácidos Grasos/metabolismo , Humanos , Insulina/metabolismo , Metabolismo de los Lípidos , Hígado/metabolismo , Ratones , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Triglicéridos/metabolismoRESUMEN
The laborious microscopic agglutination test (MAT) is the gold standard serologic test for laboratory diagnosis of leptospirosis. We developed EIA based serologic assays using recombinant proteins (rLigA, rLigB, rLipL32) and whole-cell extracts from eight Leptospira serovars as antigen and assessed the diagnostic performance of the new assay within each class, against MAT positive (MAT+) human sera panels from Portugal/PT (n = 143) and Angola/AO (n = 100). We found that a combination of recombinant proteins rLigA, rLigB and rLipL32 correctly identified antigen-specific IgG from patients with clinical and laboratory confirmed leptospirosis (MAT+) with 92% sensitivity and ~ 97% specificity (AUC 0.974) in serum from the provinces of Luanda (LDA) and Huambo (HBO) in Angola. A combination of whole cell extracts of L. interrogans sv Copenhageni (LiC), L. kirschneri Mozdok (LkM), L. borgpetersenii Arborea (LbA) and L. biflexa Patoc (LbP) accurately identified patients with clinical and laboratory confirmed leptospirosis (MAT+) with 100% sensitivity and ~ 98% specificity for all provinces of Angola and Portugal (AUC: 0.997 for AO/LDA/HBO, 1.000 for AO/HLA, 0.999 for PT/AZ and 1.000 for PT/LIS). Interestingly, we found that MAT+ IgG+ serum from Angola had a significantly higher presence of IgD and that IgG3/IgG1 isotypes were significantly increased in the MAT+ IgG+ serum from Portugal. Given that IgM/IgD class and IgG3/IgG1 specific isotypes are produced in the earliest course of infection, immunoglobulin G isotyping may be used to inform diagnosis of acute leptospirosis. The speed, ease of use and accuracy of EIA tests make them excellent alternatives to the laborious and expensive MAT for screening acute infection in areas where circulating serovars of pathogenic Leptospira are well defined.
Asunto(s)
Leptospira , Leptospirosis , Enfermedad Aguda , Pruebas de Aglutinación , Anticuerpos Antibacterianos , Antígenos Bacterianos , Extractos Celulares , Ensayo de Inmunoadsorción Enzimática , Humanos , Técnicas para Inmunoenzimas , Inmunoglobulina D , Inmunoglobulina G , Proteínas Recombinantes , Pruebas SerológicasRESUMEN
Leptospirosis is a widespread zoonotic disease caused by pathogenic spirochetes of the genus Leptospira. The commercially available vaccines are bacterins that offer limited protection, short-term effect, and serovar-specific immunity. The development of novel immunization strategies is crucial to control the infection and decrease the chances of new outbreaks. In this study, purified monoclonal antibodies (mAbs) anti-LipL32 (1D9 and mAb3) were evaluated by their capacity to bind and neutralize the pathogen improving host survival. For that, an in vitro growth inhibition assay, and in vivo passive immunization were performed in animal model. Syrian hamsters were passively immunized by three different strategies. Hamsters immunized with mAb3 6 h prior to the lethal challenge showed a significantly higher survival rate of 61.1%, and a significant reduction in tissue damage in the lungs. Cumulatively, our results showed that anti-LipL32 mAbs inhibited the growth of L. interrogans in vitro, and that passive immunization offered significant protection in animal model when administered prior to infection.
Asunto(s)
Anticuerpos Antibacterianos/inmunología , Anticuerpos Monoclonales/inmunología , Proteínas de la Membrana Bacteriana Externa/inmunología , Leptospira interrogans/inmunología , Leptospirosis/prevención & control , Lipoproteínas/inmunología , Animales , Anticuerpos Antibacterianos/farmacología , Anticuerpos Monoclonales/farmacología , Cricetinae , Modelos Animales de Enfermedad , Femenino , Inmunización , Leptospirosis/microbiología , Leptospirosis/mortalidad , Leptospirosis/patología , Resultado del TratamientoRESUMEN
BACKGROUND: Leptospirosis is an emerging zoonosis attributed to multiple reservoirs. Climatic conditions influence the transmission of pathogenic leptospires, which require warm and humid conditions for survival. The influence of seasonality in human and animal leptospirosis in the subtropical region of Brazil remains poorly understood. METHODS: We performed a retrospective study to describe the patterns of human and animal exposure to leptospirosis and their association with precipitation events in Southern Brazil. Rainfall data were obtained from satellite images. Serum samples were tested using the microscopic agglutination test (MAT); samples with titer ≥ 100 were defined as seroreactive. Linear regression and Pearson's correlation were performed to assess whether there is a relationship between these variables. RESULTS: We found that precipitation events were not significantly associated with the exposure to leptospirosis in humans or animal species, except for dogs. The interspecies analysis revealed an association between canine and human exposure to leptospirosis. Leptospira kirschneri serovar Butembo (serogroup Autumnalis) presented the highest seroreactivity in humans. CONCLUSION: This study provides valuable insights in human and animal leptospirosis in Southern Brazil. These insights will be essential to design intervention measures directed to reduce disease dissemination.
Asunto(s)
Leptospirosis/epidemiología , Zoonosis/epidemiología , Animales , Anticuerpos Antibacterianos/sangre , Brasil/epidemiología , Perros , Humanos , Leptospirosis/veterinaria , Estudios Retrospectivos , Factores de RiesgoRESUMEN
Enzootic Pneumonia (EP) is caused by the Mycoplasma hyopneumoniae pathogenic bacteria, and it represents a significant respiratory disease that is responsible for major economic losses within the pig industry throughout the world. The bacterins that are currently commercially available have been proven to offer only partial protection against M. hyopneumoniae, and the development of more efficient vaccines is required. Several recombinant antigens have been evaluated via different immunization strategies and have been found to be highly immunogenic. This work describes the construction and immunological characterization of a multi-antigen chimera composed of four M. hyopneumoniae antigens: P97R1, P46, P95, and P42. Immunogenic regions of each antigen were selected and combined to encode a single polypeptide. The gene was cloned and expressed in Escherichia coli, and the chimeric protein was recognized by specific antibodies against each subunit, as well as by convalescent pig sera. The immunogenic properties of the chimera were then evaluated in a mice model through two recombinant vaccines that were formulated as follows: (1) purified chimeric protein plus adjuvant or (2) recombinant Escherichia coli bacterin. The immune response induced in BALB/c mice immunized with each formulation was characterized in terms of total IgG levels, IgG1, and IgG2a isotypes against each antigen present in the chimera. The results of the study indicated that novel chimeric protein is a potential candidate for the future development of a more effective vaccine against EP.
Asunto(s)
Antígenos Bacterianos/inmunología , Vacunas Bacterianas/inmunología , Inmunoglobulina G/sangre , Mycoplasma hyopneumoniae/inmunología , Neumonía Porcina por Mycoplasma/prevención & control , Adyuvantes Inmunológicos , Animales , Antígenos Bacterianos/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Femenino , Inmunización/veterinaria , Ratones , Ratones Endogámicos BALB C , Modelos Moleculares , Mycoplasma hyopneumoniae/genética , Neumonía Porcina por Mycoplasma/microbiología , Proteínas Recombinantes , Porcinos , Vacunas Sintéticas/inmunologíaRESUMEN
Enzootic pneumonia (EP), resulting from Mycoplasma hyopneumoniae infection is one of the most prevalent diseases in pigs and is a major cause of economic losses to the swine industry worldwide. EP is often controlled by vaccination with inactivated, adjuvanted whole-cell bacterin. However, these bacterins provide only partial protection and do not prevent M. hyopneumoniae colonization. Attempts to develop vaccines that are more efficient have made use of the recombinant DNA technology. The objective of this study was to assess the potential of recombinant M. hyopneumoniae heat shock protein P42 in vaccine preparations against EP, using piglets housed under field conditions in a M. hyopneumoniae-positive farm. The cellular and humoral immune responses were elicited after a single intramuscular inoculation of rP42 in an oil-based adjuvant, or in conjunction with whole-cell vaccine preparation. The production of INF-γ and IL-10 cytokines was quantified in the supernatant of the cultured mononuclear cells. The rP42 emulsified in oil-based adjuvant was able to trigger a strong humoral immune response. Further, it induced a cellular immune response, accompanied by the production of antibodies that reacted with the native M. hyopneumoniae protein. The rP42 mediated induction of cellular and humoral immune response in the host suggests that rP42 emulsified in an oil-based adjuvant holds promise as an effective recombinant subunit vaccine against EP.