RESUMEN
Alzheimer's disease is a devastating neurodegenerative disease with a dramatically increasing prevalence and no disease-modifying treatment. Inflammatory lifestyle factors increase the risk of developing Alzheimer's disease. Zinc deficiency is the most prevalent malnutrition in the world and may be a risk factor for Alzheimer's disease potentially through enhanced inflammation, although evidence for this is limited. Here we provide epidemiological evidence suggesting that zinc supplementation was associated with reduced risk and slower cognitive decline, in people with Alzheimer's disease and mild cognitive impairment. Using the APP/PS1 mouse model of Alzheimer's disease fed a control (35 mg/kg zinc) or diet deficient in zinc (3 mg/kg zinc), we determined that zinc deficiency accelerated Alzheimer's-like memory deficits without modifying amyloid ß plaque burden in the brains of male mice. The NLRP3-inflammasome complex is one of the most important regulators of inflammation, and we show here that zinc deficiency in immune cells, including microglia, potentiated NLRP3 responses to inflammatory stimuli in vitro, including amyloid oligomers, while zinc supplementation inhibited NLRP3 activation. APP/PS1 mice deficient in NLRP3 were protected against the accelerated cognitive decline with zinc deficiency. Collectively, this research suggests that zinc status is linked to inflammatory reactivity and may be modified in people to reduce the risk and slow the progression of Alzheimer's disease.SIGNIFICANCE STATEMENT Alzheimer's disease is a common condition mostly affecting the elderly. Zinc deficiency is also a global problem, especially in the elderly and also in people with Alzheimer's disease. Zinc deficiency contributes to many clinical disorders, including immune dysfunction. Inflammation is known to contribute to the risk and progression of Alzheimer's disease; thus, we hypothesized that zinc status would affect Alzheimer's disease progression. Here we show that zinc supplementation reduced the prevalence and symptomatic decline in people with Alzheimer's disease. In an animal model of Alzheimer's disease, zinc deficiency worsened cognitive decline because of an enhancement in NLRP3-driven inflammation. Overall, our data suggest that zinc status affects Alzheimer's disease progression, and that zinc supplementation could slow the rate of cognitive decline.
Asunto(s)
Enfermedad de Alzheimer/sangre , Disfunción Cognitiva/sangre , Progresión de la Enfermedad , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Zinc/sangre , Adulto , Anciano , Enfermedad de Alzheimer/diagnóstico por imagen , Enfermedad de Alzheimer/dietoterapia , Animales , Células Cultivadas , Disfunción Cognitiva/diagnóstico por imagen , Disfunción Cognitiva/dietoterapia , Suplementos Dietéticos , Femenino , Estudios de Seguimiento , Humanos , Mediadores de Inflamación/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Persona de Mediana Edad , Zinc/administración & dosificación , Zinc/deficienciaRESUMEN
Interleukin (IL)-1 family cytokines potently regulate inflammation, with the majority of the IL-1 family proteins being secreted from immune cells via unconventional pathways. In many cases, secretion of IL-1 cytokines appears to be closely coupled to cell death, yet the secretory mechanisms involved remain poorly understood. Here, we studied the secretion of the three best-characterized members of the IL-1 superfamily, IL-1α, IL-1ß, and IL-18, in a range of conditions and cell types, including murine bone marrow-derived and peritoneal macrophages, human monocyte-derived macrophages, HeLa cells, and mouse embryonic fibroblasts. We discovered that IL-1ß and IL-18 share a common secretory pathway that depends upon membrane permeability and can operate in the absence of complete cell lysis and cell death. We also found that the pathway regulating the trafficking of IL-1α is distinct from the pathway regulating IL-1ß and IL-18. Although the release of IL-1α could also be dissociated from cell death, it was independent of the effects of the membrane-stabilizing agent punicalagin, which inhibited both IL-1ß and IL-18 release. These results reveal that in addition to their role as danger signals released from dead cells, IL-1 family cytokines can be secreted in the absence of cell death. We propose that models used in the study of IL-1 release should be considered context-dependently.
Asunto(s)
Células de la Médula Ósea/metabolismo , Interleucina-18/metabolismo , Interleucina-1alfa/metabolismo , Interleucina-1beta/metabolismo , Macrófagos Peritoneales/metabolismo , Animales , Células de la Médula Ósea/citología , Fibroblastos/citología , Fibroblastos/metabolismo , Células HeLa , Humanos , Taninos Hidrolizables/farmacología , Macrófagos Peritoneales/citología , Ratones , Transporte de Proteínas/efectos de los fármacosRESUMEN
Infectious laryngotracheitis (ILT) is an acute respiratory disease of chickens controlled through vaccination with live-modified attenuated vaccines, the chicken embryo origin (CEO) vaccines and the tissue-culture origin (TCO) vaccines. Recently, novel recombinant vaccines have been developed using herpesvirus of turkey (HVT) and fowl pox virus (FPV) as vectors to express ILTV immunogens for protection against ILT. The objective of this study was to assess the protection efficacy against ILT induced by recombinants, live-modified attenuated, and inactivated virus vaccines when administered alone or in combination. Commercial layer pullets were vaccinated with one or more vaccines and challenged at 35 (35 WCH) or 74 weeks of age (74 WCH). Protection was assessed by scoring clinical signs; and by determining the challenge viral load in the trachea at five days post-challenge. The FPV-LT vaccinated birds were not protected when challenged at 35 weeks; the HVT-LT and TCO vaccines in combination provided protection similar to that observed in chickens vaccinated with either HVT-LT or TCO vaccines when challenged at 35 weeks, whereas protection induced by vaccination with HVT-LT followed by TCO was superior in the 74 WCH group compared with the 35 WCH group. Birds given the inactivated ILT vaccine had fewer clinical signs and/or lower viral replication at 74 WCH when combined with TCO or HVT-LT, but not when given alone. Finally, the CEO-vaccinated birds had top protection as indicated by reduction of clinical signs and viral replication when challenged at 35 weeks (74 weeks not done). These results suggest that certain vaccine combinations may be successful to produce long-term protection up to 74 weeks of age against ILT.
Asunto(s)
Pollos/inmunología , Infecciones por Herpesviridae/veterinaria , Herpesvirus Gallináceo 1/inmunología , Enfermedades de las Aves de Corral/prevención & control , Vacunación/veterinaria , Vacunas Virales/inmunología , Animales , Pollos/virología , Femenino , Virus de la Viruela de las Aves de Corral/genética , Vectores Genéticos/genética , Infecciones por Herpesviridae/prevención & control , Infecciones por Herpesviridae/virología , Herpesvirus Meleágrido 1/genética , Enfermedades de las Aves de Corral/virología , Vacunas Atenuadas/administración & dosificación , Vacunas Atenuadas/inmunología , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/inmunología , Vacunas Virales/administración & dosificaciónRESUMEN
The cochaperone BAG3 is a central protein homeostasis factor in mechanically strained mammalian cells. It mediates the degradation of unfolded and damaged forms of the actin-crosslinker filamin through chaperone-assisted selective autophagy (CASA). In addition, BAG3 stimulates filamin transcription in order to compensate autophagic disposal and to maintain the actin cytoskeleton under strain. Here we demonstrate that BAG3 coordinates protein synthesis and autophagy through spatial regulation of the mammalian target of rapamycin complex 1 (mTORC1). The cochaperone utilizes its WW domain to contact a proline-rich motif in the tuberous sclerosis protein TSC1 that functions as an mTORC1 inhibitor in association with TSC2. Interaction with BAG3 results in a recruitment of TSC complexes to actin stress fibers, where the complexes act on a subpopulation of mTOR-positive vesicles associated with the cytoskeleton. Local inhibition of mTORC1 is essential to initiate autophagy at sites of filamin unfolding and damage. At the same time, BAG3-mediated sequestration of TSC1/TSC2 relieves mTORC1 inhibition in the remaining cytoplasm, which stimulates protein translation. In human muscle, an exercise-induced association of TSC1 with the cytoskeleton coincides with mTORC1 activation in the cytoplasm. The spatial regulation of mTORC1 exerted by BAG3 apparently provides the basis for a simultaneous induction of autophagy and protein synthesis to maintain the proteome under mechanical strain.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/genética , Proteínas Reguladoras de la Apoptosis/genética , Autofagia/genética , Complejos Multiproteicos/genética , Músculo Esquelético/metabolismo , Miocitos del Músculo Liso/metabolismo , Estrés Mecánico , Serina-Treonina Quinasas TOR/genética , Citoesqueleto de Actina/metabolismo , Citoesqueleto de Actina/ultraestructura , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Secuencia de Aminoácidos , Animales , Proteínas Reguladoras de la Apoptosis/metabolismo , Fenómenos Biomecánicos , Línea Celular , Clonación Molecular , Escherichia coli/genética , Escherichia coli/metabolismo , Filaminas/genética , Filaminas/metabolismo , Expresión Génica , Regulación de la Expresión Génica , Humanos , Diana Mecanicista del Complejo 1 de la Rapamicina , Ratones , Complejos Multiproteicos/metabolismo , Músculo Esquelético/citología , Miocitos del Músculo Liso/ultraestructura , Unión Proteica , Biosíntesis de Proteínas , Ratas , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Transducción de Señal , Serina-Treonina Quinasas TOR/metabolismo , Proteína 1 del Complejo de la Esclerosis Tuberosa , Proteína 2 del Complejo de la Esclerosis Tuberosa , Proteínas Supresoras de Tumor/genética , Proteínas Supresoras de Tumor/metabolismoRESUMEN
BACKGROUND: In 2004, we initiated an inception cohort of patients with recent-onset rheumatoid arthritis (RA). Hand function was incorporated into evaluations from 2014 onward. The objectives were to examine hand function in our cohort, compare hand function with function in healthy controls and determine the factors associated with impaired function. METHODS: From February 2014 to June 2015, 139 patients (97.2 % of the cohort) had disease activity scored (28 joints, [DAS28]); the Michigan Hand Outcome Questionnaire (MHQ) and Disabilities of the Arm, Shoulder and Hand Outcome Measure (DASH) were completed, and the tip-, key- and palmar-pinch and grip strengths were measured. Sixty-nine healthy controls underwent the same evaluations. Ninety-nine patients underwent a second evaluation one year after their baseline. Descriptive statistics and linear regression models were used. Patients and controls signed informed consent. RESULTS: Patients were primarily middle-aged females with a median disease duration of 7 years; 91 patients had DAS28-remission, and 16, 23, and 9 patients had low, moderate and high disease activity, respectively. Controls scored better than did patients with (any) disease activity level; remission patients had similar DASH and key pinch function as did controls with poorer MHQ and both tip and palmar pinch and grip strength. DAS28 was consistently associated with impaired hand function. Among the patients with a one-year re-assessment, changes in DAS28 correlated (rho = 0.34 to 0.63) with changes in hand function (p ≤ 0.01 for all comparisons), but there was no correlation with palmar pinch strength. CONCLUSIONS: Disease activity was associated with hand function impairment in RA patients with variable follow-up. MHQ discriminated poorer hand function in remission patients who otherwise had similar DASH scores as the controls did.
Asunto(s)
Artritis Reumatoide/fisiopatología , Evaluación de la Discapacidad , Mano/fisiopatología , Fuerza de Pellizco , Actividades Cotidianas , Adulto , Estudios de Cohortes , Personas con Discapacidad , Femenino , Humanos , Masculino , Persona de Mediana Edad , Dimensión del Dolor , Médicos , Autoinforme , Encuestas y CuestionariosRESUMEN
Intracerebral haemorrhage (ICH) is a devastating neurovascular attack with limited treatment options. Alternative, pre-clinical modelling approaches are required to identify and trial therapeutic drug compounds. In this study we have used alginate hydrogels to model blood insult in vitro. Human whole blood was mixed with alginate and encapsulated into hydrogel beads. Beads were then incorporated in a second layer of alginate containing hyaluronic acid/chitosan nanoparticles to mimic the mechanical properties of brain tissue and create a model haematoma. Beads and model haematomas were characterised to profile size, volume, mechanical properties, release capacity and storage stability over time. Beads and model haematomas stimulate a pro-inflammatory phenotype in human monocytic and macrophage-like cells, however have no pathogenic effect on brain endothelial and neuronal cell survival or function. In conclusion, we have developed an effective strategy to model ICH in vitro, to investigate the human immune response to blood insult.
Asunto(s)
Alginatos , Hemorragia Cerebral , Hematoma , Ácido Hialurónico , Humanos , Alginatos/química , Ácido Hialurónico/química , Ácido Hialurónico/farmacología , Quitosano/química , Hidrogeles/química , Nanopartículas/química , Modelos Biológicos , Supervivencia Celular/efectos de los fármacos , Macrófagos/efectos de los fármacosRESUMEN
Interleukin-1α is a suggested dual-function cytokine that diverged from interleukin-1ß in mammals potentially by acquiring additional biological roles that relate to highly conserved regions in the pro-domain of interleukin-1α, including a nuclear localisation sequence and histone acetyltransferase-binding domains. Why evolution modified pro-interleukin-1α's subcellular location and protein interactome, and how this shaped interleukin-1α's intracellular role, is unknown. Here we show that TurboID proximity labelling with pro-interleukin-1α suggests a nuclear role for pro-interleukin-1α that involves interaction with histone acetyltransferases, including EP300. We also identify and validate inactivating mutations in the pro-interleukin-1α nuclear localisation sequence of multiple mammalian species, including toothed whales, castorimorpha and marsupials. However, histone acetyltransferase-binding domains are conserved in those species that have lost pro-interleukin-1α nuclear localisation. Together, these data suggest that histone acetyltransferase binding and nuclear localisation occurred together, and that while some species lost the nuclear localisation sequence in their pro-interleukin-1α, histone acetyltransferase binding ability was maintained. The nuclear localisation sequence was lost from several distinct species at different evolutionary times, suggesting convergent evolution, and that the loss of the nuclear localisation sequence confers some important biological outcome.
Asunto(s)
Núcleo Celular , Evolución Molecular , Interleucina-1alfa , Interleucina-1alfa/metabolismo , Interleucina-1alfa/genética , Animales , Núcleo Celular/metabolismo , Humanos , Proteína p300 Asociada a E1A/metabolismo , Proteína p300 Asociada a E1A/genética , Histona Acetiltransferasas/metabolismo , Histona Acetiltransferasas/genética , Señales de Localización Nuclear/genética , Señales de Localización Nuclear/metabolismo , Unión Proteica , Secuencia de AminoácidosRESUMEN
Unlike mammals, regenerative model organisms such as amphibians and fish are capable of spinal cord regeneration after injury. Certain key differences between regenerative and nonregenerative organisms have been suggested as involved in promoting this process, such as the capacity for neurogenesis and axonal regeneration, which appear to be facilitated by favorable astroglial, inflammatory and immune responses. These traits provide a regenerative-permissive environment that the mammalian spinal cord appears to be lacking. Evidence for the regenerative nonpermissive environment in mammals is given by the fact that they possess neural stem/progenitor cells, which transplanted into permissive environments are able to give rise to new neurons, whereas in the nonpermissive spinal cord they are unable to do so. We discuss the traits that are favorable for regeneration, comparing what happens in mammals with each regenerative organism, aiming to describe and identify the key differences that allow regeneration. This comparison should lead us toward finding how to promote regeneration in organisms that are unable to do so.
Asunto(s)
Regeneración Nerviosa , Médula Espinal/fisiología , Animales , Axones/fisiología , Humanos , Mamíferos , Neurogénesis , Traumatismos de la Médula Espinal/inmunología , Traumatismos de la Médula Espinal/patología , Traumatismos de la Médula Espinal/fisiopatologíaRESUMEN
Aicardi-Goutières syndrome (AGS1-9) is a genetically determined encephalopathy that falls under the type I interferonopathy disease class, characterized by excessive type I interferon (IFN-I) activity, coupled with upregulation of IFN-stimulated genes (ISGs), which can be explained by the vital role these proteins play in self-non-self-discrimination. To date, few mouse models fully replicate the vast clinical phenotypes observed in AGS patients. Therefore, we investigated the use of zebrafish as an alternative species for generating a clinically relevant model of AGS. Using CRISPR-cas9 technology, we generated a stable mutant zebrafish line recapitulating AGS5, which arises from recessive mutations in SAMHD1. The resulting homozygous mutant zebrafish larvae possess a number of neurological phenotypes, exemplified by variable, but increased expression of several ISGs in the head region, a significant increase in brain cell death, microcephaly and locomotion deficits. A link between IFN-I signaling and cholesterol biosynthesis has been highlighted by others, but not previously implicated in the type I interferonopathies. Through assessment of neurovascular integrity and qPCR analysis we identified a significant dysregulation of cholesterol biosynthesis in the zebrafish model. Furthermore, dysregulation of cholesterol biosynthesis gene expression was also observed through RNA sequencing analysis of AGS patient whole blood. From this novel finding, we hypothesize that cholesterol dysregulation may play a role in AGS disease pathophysiology. Further experimentation will lend critical insight into the molecular pathophysiology of AGS and the potential links involving aberrant type I IFN signaling and cholesterol dysregulation.
Asunto(s)
Enfermedades Autoinmunes del Sistema Nervioso , Interferón Tipo I , Malformaciones del Sistema Nervioso , Animales , Ratones , Enfermedades Autoinmunes del Sistema Nervioso/genética , Enfermedades Autoinmunes del Sistema Nervioso/metabolismo , Interferón Tipo I/genética , Interferón Tipo I/metabolismo , Malformaciones del Sistema Nervioso/genética , Malformaciones del Sistema Nervioso/metabolismo , Proteína 1 que Contiene Dominios SAM y HD/genética , Pez Cebra/genética , Pez Cebra/metabolismoRESUMEN
Viral arthritis/tenosynovitis, a disease caused by avian reovirus (ARV), leads to great economic losses for the chicken industry worldwide. Since autumn 2011, the poultry industries in the United States and Canada have sustained significant economic losses in the progeny of broiler breeders vaccinated with classic strains of ARV. Vaccination failure has been caused by field challenge with variant ARVs. The variant field ARVs are refractory to the immunity stimulated by classic vaccines and have become the prevalent challenge in the field. Because all genotypes described in the literature have been reported to be circulating in Canada, genotyping of circulating ARVs is paramount for the selection of appropriate isolates, representative of the field challenge, for use in autogenous vaccines. In this review, the history of ARVs and the current situation in Canada are discussed. On the basis of recent field data, inadequate measures commonly used in the field are discussed, and successful vaccination strategies are recommended.
Estudio recapitulativo- Revisión de la artritis viral en Canadá La artritis/tenosinovitis viral, una enfermedad causada por el reovirus aviares (ARV), genera grandes pérdidas económicas para la industria avícola en todo el mundo. Desde el otoño del 2011, las industrias avícolas de los Estados Unidos y Canadá han sufrido pérdidas económicas significativas en la progenie de reproductoras de pollos de engorde vacunadas con cepas clásicas de reovirus aviares. Las fallas de la vacunación han sido causadas por el desafío de campo con reovirus aviares variantes. Los reovirus aviares de campo variantes son refractarios a la inmunidad estimulada por las vacunas clásicas y se han convertido en el desafío predominante en el campo. Debido a que se ha reportado que todos los genotipos descritos en la literatura están circulando en Canadá, la determinación del genotipo de los reovirus aviares circulantes es fundamental para la selección de aislamientos apropiados, representativos del desafío de campo, para su uso en vacunas autógenas. En esta revisión, se discute la historia de los reovirus aviares y la situación actual en Canadá. Sobre la base de datos de campo recientes, se analizan las medidas inadecuadas comúnmente utilizadas en el campo y se recomiendan estrategias de vacunación exitosas.
Asunto(s)
Artritis Infecciosa , Orthoreovirus Aviar , Enfermedades de las Aves de Corral , Infecciones por Reoviridae , Vacunas Virales , Animales , Pollos , Infecciones por Reoviridae/epidemiología , Infecciones por Reoviridae/veterinaria , Enfermedades de las Aves de Corral/epidemiología , Enfermedades de las Aves de Corral/prevención & control , Filogenia , Artritis Infecciosa/veterinaria , Canadá/epidemiologíaRESUMEN
Despite the global health burden, treatment of spontaneous intracerebral haemorrhage (ICH) is largely supportive, and translation of specific medical therapies has not been successful. Zebrafish larvae offer a unique platform for drug screening to rapidly identify neuroprotective compounds following ICH. We applied the Spectrum Collection library compounds to zebrafish larvae acutely after ICH to screen for decreased brain cell death and identified 150 successful drugs. Candidates were then evaluated for possible indications with other cardiovascular diseases. Six compounds were identified, including two angiotensin-converting enzyme inhibitors (ACE-Is). Ramipril and quinapril were further assessed to confirm a significant 55% reduction in brain cell death. Proteomic analysis revealed potential mechanisms of neuroprotection. Using the INTERACT2 clinical trial dataset, we demonstrated a significant reduction in the adjusted odds of an unfavourable shift in the modified Rankin scale at 90â days for patients receiving an ACE-I after ICH (versus no ACE-I; odds ratio, 0.80; 95% confidence interval, 0.68-0.95; P=0.009). The zebrafish larval model of spontaneous ICH can be used as a reliable drug screening platform and has identified therapeutics that may offer neuroprotection. This article has an associated First Person interview with the first author of the paper.
Asunto(s)
Neuroprotección , Pez Cebra , Animales , Hemorragia Cerebral/tratamiento farmacológico , Evaluación Preclínica de Medicamentos , Humanos , Larva , ProteómicaRESUMEN
The PQBP1 (polyglutamine tract-binding protein 1) gene encodes a nuclear protein that regulates pre-mRNA splicing and transcription. Mutations in the PQBP1 gene were reported in several X chromosome-linked mental retardation disorders including Golabi-Ito-Hall syndrome. The missense mutation that causes this syndrome is unique among other PQBP1 mutations reported to date because it maps within a functional domain of PQBP1, known as the WW domain. The mutation substitutes tyrosine 65 with cysteine and is located within the conserved core of aromatic amino acids of the domain. We show here that the binding property of the Y65C-mutated WW domain and the full-length mutant protein toward its cognate proline-rich ligands was diminished. Furthermore, in Golabi-Ito-Hall-derived lymphoblasts we showed that the complex between PQBP1-Y65C and WBP11 (WW domain-binding protein 11) splicing factor was compromised. In these cells a substantial decrease in pre-mRNA splicing efficiency was detected. Our study points to the critical role of the WW domain in the function of the PQBP1 protein and provides an insight into the molecular mechanism that underlies the X chromosome-linked mental retardation entities classified globally as Renpenning syndrome.
Asunto(s)
Proteínas Portadoras/química , Proteínas Portadoras/genética , Mutación Missense , Proteínas Nucleares/química , Proteínas Nucleares/genética , Empalme Alternativo , Calorimetría/métodos , Dicroismo Circular , Proteínas de Unión al ADN , Humanos , Discapacidad Intelectual/genética , Ligandos , Linfocitos/metabolismo , Espectroscopía de Resonancia Magnética , Mutación , Prolina/química , Estructura Terciaria de Proteína , Resonancia por Plasmón de Superficie , Transcripción GenéticaRESUMEN
The growing demand for binding assays to study protein-protein interaction can be addressed by peptide array-based methods. The SPOT technique is a widespread peptide-array technology, which is able to distinguish semi-quantitatively the binding affinities of peptides to defined protein targets within one array. The quality of an assay system used for probing peptide arrays depends on the well-balanced combination of screening and read-out methods. The former address the steady-state of analyte capture, whereas the latter provide the means to detect captured analyte. In all cases, however, false-positive results can occur when challenging a peptide array with analyte or detecting captured analyte with label conjugates. Little is known about the cross-reactivity of peptides with the detection agents. Here, we describe at the amino acid level the potential of (i) 5-(and 6)-carboxytetramethylrhodamine (5(6)-TAMRA), (ii) fluoresceinisothiocyanate in form of the peptide-bound fluorescein-substituted thiourea derivative (FITC), and (iii) biotin/streptavidin-POD to cross-react with individual amino acids in a peptide sequence.
Asunto(s)
Aminoácidos/análisis , Bioensayo/métodos , Celulosa/química , Péptidos/química , Secuencia de Aminoácidos , Datos de Secuencia Molecular , Péptidos/genética , Unión ProteicaRESUMEN
Hemorrhagic enteritis virus (HEV) is an immunosuppressive adenovirus that causes an acute clinical disease characterized by hemorrhagic gastroenteritis in 4-week-old turkeys and older. Recurrent incidence of secondary infections (e.g., systemic bacterial infections, cellulitis, and elevated mortality), may be associated with the presence of field-type HEV in Canadian turkey farms. We speculate that field-type HEV and vaccine/vaccine-like strains can be differentiated through analysis of the viral genomes, hexon genes, and the specific virulence factors (e.g., ORF1, E3, and fib knob domain). Nine out of sixteen spleens obtained from cases suspected of immunosuppression by HEV were analyzed. The limited data obtained showed that: (1) field-type HEV circulates in many non-vaccinated western Canadian flocks; (2) field-type HEV circulates in vaccinated flocks with increased recurrent bacterial infections; and (3) the existence of novel point mutations in hexon, ORF1, E3, and specially fib knob domains. This is the first publication showing the circulation of wild-type HEV in HEV-vaccinated flocks in Western Canada, and the usefulness of a novel procedure that allows whole genome sequencing of HEV directly from spleens, without passaging in cell culture or passaging in vivo. Further studies focusing more samples are required to confirm our observations and investigate possible vaccination failure.
Asunto(s)
Infecciones por Adenoviridae/veterinaria , Genoma Viral , Enfermedades de las Aves de Corral/virología , Siadenovirus/genética , Pavos/virología , Infecciones por Adenoviridae/epidemiología , Infecciones por Adenoviridae/virología , Proteínas E3 de Adenovirus/química , Proteínas E3 de Adenovirus/genética , Vacunas contra el Adenovirus/inmunología , Animales , Canadá/epidemiología , Proteínas de la Cápside/química , Proteínas de la Cápside/genética , Genes Virales , Glicosilación , Mutación , Sistemas de Lectura Abierta , Siadenovirus/inmunología , Siadenovirus/aislamiento & purificación , Siadenovirus/patogenicidad , Bazo/virología , Proteínas Virales/genética , Factores de Virulencia/genética , Secuenciación Completa del GenomaRESUMEN
In this study, we aimed to molecularly characterize 14 whole genome sequences of chicken astrovirus (CAstV) isolated from samples obtained from white chick syndrome (WCS) outbreaks in Western Canada during the period of 2014-2019. Genome sequence comparisons showed all these sequences correspond to the novel Biv group from which no confirmed representatives were published in GenBank. Molecular recombination analyses using recombination detection software (i.e., RDP5 and SimPlot) and phylogenetic analyses suggest multiple past recombination events in open reading frame (ORF)1a, ORF1b, and ORF2. Our findings suggest that recombination events and the accumulation of point mutations may have contributed to the substantial genetic variation observed in CAstV and evidenced by the current seven antigenic sub-clusters hitherto described. This is the first paper that describes recombination events in CAstV following analysis of complete CAstV sequences originated in Canada.
Asunto(s)
Infecciones por Astroviridae/veterinaria , Infecciones por Astroviridae/virología , Avastrovirus/genética , Pollos/virología , Enfermedades de las Aves de Corral/virología , Recombinación Genética , Animales , Infecciones por Astroviridae/epidemiología , Infecciones por Astroviridae/patología , Avastrovirus/clasificación , Secuencia de Bases , Canadá/epidemiología , Genoma Viral , Genotipo , Hígado/patología , Epidemiología Molecular , Sistemas de Lectura Abierta , Filogenia , Enfermedades de las Aves de Corral/epidemiología , Enfermedades de las Aves de Corral/patologíaRESUMEN
Infectious laryngotracheitis virus (ILTV) causes an acute upper respiratory disease in chickens called infectious laryngotracheitis (ILT). Live attenuated vaccines are effective in disease control; however, they have residual virulence, which makes them able to replicate, cause disease and revert to the original virulent form. Information is scarce on the molecular nature of ILTV that is linked to ILT in Canada. This study aims to determine whether isolates originating from ILT cases in Western Canada are a wild type or vaccine origin. Samples submitted for the diagnosis of ILT between 2009-2018 were obtained from Alberta (AB, n = 46) and British Columbia (BC, n = 9). For genotyping, a Sanger sequencing of open reading frame (ORF) a and b was used. A total of 27 from AB, and 5 from BC samples yielded a fragment of 1751 base pairs (bp). Three of the BC samples classified as group IV (CEO vaccine strains) and 2 as group V (CEO revertant). Of the AB samples, 22 samples clustered with group V, 3 with group VI (wild type), and 2 with group VII, VIII, and IX (wild type). Overall, 17 non-synonymous single nucleotide polymorphisms (SNPs) were detected. Further studies are underway to ascertain the virulence and transmission potential of these isolates.
RESUMEN
The NLRP3 inflammasome is an important regulator of the sterile inflammatory response, and its activation by host-derived sterile molecules leads to the intracellular activation of caspase-1, processing of the pro-inflammatory cytokines interleukin-1ß (IL-1ß)/IL-18, and pyroptotic cell death. Inappropriate activation of NLRP3 drives a chronic inflammatory response and is implicated in several non-communicable diseases, including gout, atherosclerosis, typeâ II diabetes and Alzheimer's disease. In this study, we report the design, synthesis and biological evaluation of novel boron compounds (NBCs) as NLRP3 inflammasome inhibitors. Structure-activity relationships (SAR) show that 4-fluoro substituents on the phenyl rings retain NLRP3 inhibitory activity, whereas more steric and lipophilic substituents diminish activity. Loss of inhibitory activity is also observed if the CCl3 group on the oxazaborine ring is replaced by a CF3 group. These findings provide additional understanding of the NBC series and will aid in the development of these NLRP3 inhibitors as tool compounds or therapeutic candidates for sterile inflammatory diseases.
Asunto(s)
Antiinflamatorios/síntesis química , Compuestos de Boro/química , Diseño de Fármacos , Inflamasomas/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/antagonistas & inhibidores , Animales , Antiinflamatorios/química , Antiinflamatorios/farmacología , Células de la Médula Ósea/citología , Compuestos de Boro/síntesis química , Compuestos de Boro/farmacología , Células Cultivadas , Interleucina-1beta/metabolismo , Lipopolisacáridos/farmacología , Macrófagos/citología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Ratones , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Relación Estructura-ActividadRESUMEN
Viral Arthritis (VA), a disease caused by Avian Reovirus (ARV), has emerged as a significant cause of economic losses in broiler chicken flocks in Western Canada. These outbreaks were characterized by 4-13% morbidity, followed by a spike in mortality/culling that in extreme cases required total flock depopulation. From 2012-2017, 38 ARV isolates were recovered. Molecular characterization of a partial segment of the sigma (σ)C gene shows all six previously known ARV clusters in Western Canadian broiler chickens. The most numerous clusters were Cluster#4 and Cluster #5 while the most variable clusters were Cluster#1 (76.7-100% identity), Cluster#2 (66-99.3%), and Cluster#4 (62-100%). This variation suggests that an autogenous vaccine may not protect against a same-cluster challenge virus. This is the first publication showing the wide genetic diversity of ARV Cluster#4, the circulation of all six worldwide reported ARV clusters in Canada, and important differences in ARV Cluster classification among researchers.
Asunto(s)
Artritis Infecciosa/veterinaria , Variación Genética , Orthoreovirus Aviar/genética , Orthoreovirus Aviar/aislamiento & purificación , Enfermedades de las Aves de Corral/virología , Infecciones por Reoviridae/veterinaria , Animales , Artritis Infecciosa/virología , Canadá/epidemiología , Pollos , Análisis por Conglomerados , Brotes de Enfermedades , Epidemiología Molecular , Orthoreovirus Aviar/clasificación , Filogenia , Enfermedades de las Aves de Corral/epidemiología , Homología de Secuencia , Proteínas Virales/genéticaRESUMEN
Commercial broiler and layer chickens are heavily vaccinated against economically important viral diseases with a view of preventing morbidity, mortality, and production impacts encountered during short production cycles. Hatchery vaccination is performed through in ovo embryo vaccination prehatch or spray and subcutaneous vaccinations performed at the day of hatch before the day-old chickens are being placed in barns with potentially contaminated environments. Commercially, multiple vaccines (e.g., live, live attenuated, and viral vectored vaccines) are available to administer through these routes within a short period (embryo day 18 prehatch to day 1 posthatch). Although the ability to mount immune response, especially the adaptive immune response, is not optimal around the hatch, it is possible that the efficacy of these vaccines depends partly on innate host responses elicited in response to replicating vaccine viruses. This review focuses on the current knowledge of hatchery vaccination in poultry and potential mechanisms of hatchery vaccine-mediated protective responses and limitations.
Asunto(s)
Pollos/inmunología , Enfermedades de las Aves de Corral/prevención & control , Vacunación/veterinaria , Vacunas Virales/administración & dosificación , Virosis/veterinaria , Animales , Embrión de Pollo , Pollos/virología , Inmunización Pasiva , Enfermedades de las Aves de Corral/inmunología , Enfermedades de las Aves de Corral/virología , Vacunación/normas , Vacunas Atenuadas/administración & dosificación , Vacunas Atenuadas/inmunología , Vacunas Combinadas/administración & dosificación , Vacunas Virales/inmunología , Virosis/inmunología , Virosis/prevención & control , Virosis/virologíaRESUMEN
Cytosine-guanosine deoxynucleotides (CpG) DNA can be delivered in ovo at embryo day (ED)18 for the stimulation of toll-like receptor (TLR)21 signaling pathway that ultimately protects chickens against a number of bacterial and viral infections. There is a dearth of information understanding the mechanisms of protection induced by in ovo delivered CpG DNA. The objective of this study was to determine the immune cell changes post-hatch following in ovo delivery of the TLR21 ligand, CpG DNA. In order to quantify changes of percentage of KUL01+, IgM+ B, cluster of differentiation (CD)4+ and CD8α+ cells, trachea, lung, duodenum, large intestine, spleen and bursa of Fabricius were collected on day 1 post-hatch. We found increased recruitments of KUL01+ cells, in organs of these body systems post-hatch following in ovo delivery of CpG DNA. Although IgM+ B cells, CD4+ and CD8α+ cells were increased in lungs and immune system organs, these cells were not quantifiable from the trachea, duodenum and large intestine immediately following the hatch. Furthermore, when CpG DNA is delivered in ovo and subsequently infected with infectious laryngotracheitis virus (ILTV) post-hatch on day 1, CpG DNA reduces morbidity and mortality resulting from ILTV infection. This study provides insights into the mechanisms of host responses elicited following in ovo delivery of CpG DNA in avian species.