RESUMEN
Breakdown of the blood-central nervous system barrier (BCNSB) is a hallmark of many neuroinflammatory disorders, such as multiple sclerosis (MS). Using a mouse model of MS, experimental autoimmune encephalomyelitis (EAE), we show that endothelial-to-mesenchymal transition (EndoMT) occurs in the CNS before the onset of clinical symptoms and plays a major role in the breakdown of BCNSB function. EndoMT can be induced by an IL-1ß-stimulated signaling pathway in which activation of the small GTPase ADP ribosylation factor 6 (ARF6) leads to crosstalk with the activin receptor-like kinase (ALK)-SMAD1/5 pathway. Inhibiting the activation of ARF6 both prevents and reverses EndoMT, stabilizes BCNSB function, reduces demyelination, and attenuates symptoms even after the establishment of severe EAE, without immunocompromising the host. Pan-inhibition of ALKs also reduces disease severity in the EAE model. Therefore, multiple components of the IL-1ß-ARF6-ALK-SMAD1/5 pathway could be targeted for the treatment of a variety of neuroinflammatory disorders.
Asunto(s)
Encefalomielitis Autoinmune Experimental , Proteínas de Unión al GTP Monoméricas , Esclerosis Múltiple , Receptores de Activinas/metabolismo , Animales , Sistema Nervioso Central/metabolismo , Ratones , Ratones Endogámicos C57BL , Proteínas de Unión al GTP Monoméricas/metabolismo , Enfermedades Neuroinflamatorias , Proteínas Tirosina Quinasas Receptoras/metabolismo , Transducción de SeñalRESUMEN
Macrophages chronically infected with HIV-1 serve as a reservoir that contributes to HIV-1 persistence during antiretroviral therapy; however, the mechanisms governing the establishment and maintenance of this virus reservoir have not been fully elucidated. Here, we show that HIV-1 enters a state reminiscent of latency in monocyte-derived macrophages (MDMs), characterized by integrated proviral DNA with decreased viral transcription. This quiescent state is associated with decreased NF-κB p65, RNA polymerase II, and p-TEFb recruitment to the HIV-1 promoter as well as maintenance of promoter chromatin in a transcriptionally nonpermissive state. MDM transition to viral latency is mediated by type I IFN signaling, as inhibiting type I IFN signaling or blocking type 1 IFN prevents the establishment of latent infection. Knockdown studies demonstrate that the innate immune signaling molecule mitochondrial antiviral signaling protein (MAVS) is required for the transition to latency. Finally, we demonstrate a role for the viral accessory protein Vpr in the establishment of HIV-1 latency in macrophages. Our data indicate that HIV-1-induced type I IFN production is responsible for the establishment of viral latency in MDMs and identify possible therapeutic targets for the prevention or elimination of this important HIV-1 reservoir.
Asunto(s)
Infecciones por VIH , VIH-1 , Interferón Tipo I , Macrófagos , Latencia del Virus , Humanos , Cromatina , Infecciones por VIH/inmunología , Macrófagos/metabolismo , Macrófagos/virología , FN-kappa B/metabolismo , Factor B de Elongación Transcripcional Positiva/genética , ARN Polimerasa II , Activación Viral , Interferón Tipo I/inmunologíaRESUMEN
Industrial hemp (Cannabis sativa) is an emerging crop in Arizona, with many uses, including fiber, cosmetic products, and health food. In 2020, severe curly top disease outbreaks were observed in several hemp fields in Yuma and Graham Counties, Arizona, where disease incidence and severity were considerably high, up to 100% crop loss occurring in some fields. A wide range of symptoms have been observed at different infection stages and plant growth stages at the time of infection. Early stage symptoms manifest as light green-to-yellowing of new growth, similar to sulfur or micronutrient deficiency, usually combined with older leaves with dark green "blotchy" mosaic mottling overlaying light green chlorosis. Mosaic mottling of older leaves continues into mid-growth stage, and is coupled with more severe yellowing and witch's broom (stunted leaves and shortened internode length of stem) of apical meristematic tissue. Curling and twisting of new leaves has also been observed. Symptoms often appear to be isolated to individual branches, with other branches showing no visual symptoms, often outgrowing and covering affected branches until harvest. Late stage symptoms include severe leaf curling with or without twisting, continued stunting, and necrosis of yellow leaves, resulting in significant yield reduction. Severely affected plants dwarfed by the virus experienced high mortality rates later into the season, most likely attributed to reduced ability to overcome abiotic stress conditions. These symptoms indicated the likelihood of curly top caused by Beet curly top virus (BCTV), which has been recently reported in Colorado (Giladi et al., 2020). Shoots were collected from thirty-eight symptomatic and nine asymptomatic hemp plants from July to August, 2020. Leaves were also collected as positive control from four chili pepper plants with or without curly top symptoms in Cochise County. Genomic DNA was extracted using DNeasy Plant Pro Kit (Qiagen Inc., Valencia, CA) according to the manufacturer's instructions. BCTV-specific primers BCTV1 and BCTV2 were used to detect BCTV following a method by Rondon (Rondon et al., 2016). A 500 bp DNA fragment, indicative of BCTV, was amplified from all symptomatic hemp and chili pepper samples, but not from asymptomatic samples. Sequence analysis of this 500 bp DNA fragment revealed 98.99 % identity with GenBank accession MK803280, which is Beet curly top virus isolate from hemp identified in Western Colorado (Giladi et al., 2020). The full-length genomes of BCTV isolates from hemp and chili peppers were generated with additional primers 328F/945R (620bp), 455F/ 945R (490bp), OutR/ 2213F (1,190bp), 2609R/ 1278R (1,340bp), BCTV2/ 2609R (1,890bp) (Rondon et al., 2016, Strausbaugh et al., 2008). The complete nucleotide sequence (MW182244) from hemp was 2,929 bp and had 99.35% sequence identity with GenBank accession KX867055, which was a Worland strain of Beet curly top virus isolated from an Idaho sugar beet plant (Strausbaugh et al., 2017). Our hemp BCTV genome sequences shared 96.08% identity with the hemp strain of BCTV from Colorado (MK803280) and 99.50% identity with the BCTV isolate (MW188519) from chili pepper identified in this study. BCTV was reported on outdoor hemp in Western Colorado, in 2020 (Giladi et al., 2020). This is the first report of BCTV in Arizona causing curly top of industrial hemp in the field. In Arizona, BCTV is widespread on many agronomic crops including chili peppers and spread primarily by the phloem-feeding beet leafhoppers: Circulifer tenellus (Hemiptera: Cicadellidae) (Bennett, 1967). Due to the wide distribution of beet leafhoppers and abundant range of host plants for the virus, BCTV may become one of the most yield-limiting factors affecting the emerging industrial hemp production systems in Arizona.
RESUMEN
BACKGROUND: Treatments for autoimmune diseases aim to dampen autoreactivity while preserving normal immune function. In CD4+ T cells, the transcription factor Oct1/Pou2f1 is a dispensable transcription factor for T cell development and response to primary infection, but promotes expression of target genes, including Il2 and Ifng, under conditions of antigen reencounter. As a result, they are more strongly expressed upon secondary stimulation. Such repeated antigen encounters occur in memory recall responses, in autoimmunity where self-antigen can be recognized multiple times, and in chronic infection where foreign antigen is persistent. Based on these previous findings, we hypothesized that Oct1 loss would protect animals from autoimmunity but maintain normal responses to pathogens in the CNS. OBJECTIVE: We used a conditional mouse Oct1 (Pou2f1) allele and a CD4-Cre driver to determine the effect of T cell-specific Oct1 loss on autoimmune- and viral-induced neuroinflammation using an autoantigen-driven EAE model of autoimmunity and a JHMV model of viral infection. RESULTS: Oct1 conditional deletion mitigated clinical scores and reduced infiltrating T cells and cytokine production in the EAE model. Consistently, Oct1-deficient CD4+ T cells stimulated in vitro showed increased expression of markers associated with T cell anergy, particularly in the absence of co-stimulatory signals. In contrast, anti-viral T cell effector functions are intact in the absence of Oct1, with no changes in neuroinflammation, infiltrating T cells or cytokine production. CONCLUSION: Our findings uncover a significant difference between the effect of Oct1 loss on autoimmune and anti-pathogen responses, which potentially could be exploited for therapeutic benefit.
Asunto(s)
Autoinmunidad/fisiología , Linfocitos T CD4-Positivos/metabolismo , Encefalomielitis Autoinmune Experimental/metabolismo , Mediadores de Inflamación/metabolismo , Factores de Crecimiento Nervioso/metabolismo , Factor 1 de Transcripción de Unión a Octámeros/deficiencia , Secuencia de Aminoácidos , Animales , Linfocitos T CD4-Positivos/inmunología , Encefalomielitis Autoinmune Experimental/genética , Encefalomielitis Autoinmune Experimental/inmunología , Inflamación/genética , Inflamación/inmunología , Inflamación/metabolismo , Mediadores de Inflamación/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Factores de Crecimiento Nervioso/genética , Factores de Crecimiento Nervioso/inmunología , Factor 1 de Transcripción de Unión a Octámeros/genética , Factor 1 de Transcripción de Unión a Octámeros/inmunologíaRESUMEN
Symbiotic microbes impact the function and development of the central nervous system (CNS); however, little is known about the contribution of the microbiota during viral-induced neurologic damage. We identify that commensals aid in host defense following infection with a neurotropic virus through enhancing microglia function. Germfree mice or animals that receive antibiotics are unable to control viral replication within the brain leading to increased paralysis. Microglia derived from germfree or antibiotic-treated animals cannot stimulate viral-specific immunity and microglia depletion leads to worsened demyelination. Oral administration of toll-like receptor (TLR) ligands to virally infected germfree mice limits neurologic damage. Homeostatic activation of microglia is dependent on intrinsic signaling through TLR4, as disruption of TLR4 within microglia, but not the entire CNS (excluding microglia), leads to increased viral-induced clinical disease. This work demonstrates that gut immune-stimulatory products can influence microglia function to prevent CNS damage following viral infection.
Asunto(s)
Encefalitis Viral/patología , Encefalitis Viral/prevención & control , Microbioma Gastrointestinal/inmunología , Microglía/inmunología , Transducción de Señal , Simbiosis , Receptores Toll-Like/metabolismo , Animales , Modelos Animales de Enfermedad , Vida Libre de Gérmenes , RatonesRESUMEN
CXCR2 is a chemokine receptor expressed on oligodendroglia that has been implicated in the pathogenesis of neuroinflammatory demyelinating diseases as well as enhancement of the migration, proliferation, and myelin production by oligodendroglia. Using an inducible proteolipid protein (Plp) promoter-driven Cre-loxP recombination system, we were able to assess how timed ablation of Cxcr2 in oligodendroglia affected disease following intracranial infection with the neurotropic JHM strain of mouse hepatitis virus (JHMV). Generation of Plp-Cre-ER(T)::Cxcr2flox/flox transgenic mice (termed Cxcr2-CKO mice) allows for Cxcr2 to be silenced in oligodendrocytes in adult mice following treatment with tamoxifen. Ablation of oligodendroglia Cxcr2 did not influence clinical severity in response to intracranial infection with JHMV. Infiltration of activated T cells or myeloid cells into the central nervous system (CNS) was not affected, nor was the ability to control viral infection. In addition, the severity of demyelination was similar between tamoxifen-treated mice and vehicle-treated controls. Notably, deletion of Cxcr2 resulted in increased remyelination, as assessed by g-ratio (the ratio of the inner axonal diameter to the total outer fiber diameter) calculation, compared to that in vehicle-treated control mice. Collectively, our findings argue that CXCR2 signaling in oligodendroglia is dispensable with regard to contributing to neuroinflammation, but its deletion enhances remyelination in a preclinical model of the human demyelinating disease multiple sclerosis (MS).IMPORTANCE Signaling through the chemokine receptor CXCR2 in oligodendroglia is important for developmental myelination in rodents, while chemical inhibition or nonspecific genetic deletion of CXCR2 appears to augment myelin repair in animal models of the human demyelinating disease multiple sclerosis (MS). To better understand the biology of CXCR2 signaling on oligodendroglia, we generated transgenic mice in which Cxcr2 is selectively ablated in oligodendroglia upon treatment with tamoxifen. Using a viral model of neuroinflammation and demyelination, we demonstrate that genetic silencing of CXCR2 on oligodendroglia did not affect clinical disease, neuroinflammation, or demyelination, yet there was increased remyelination. These findings support and extend previous findings suggesting that targeting CXCR2 may offer a therapeutic avenue for enhancing remyelination in patients with demyelinating diseases.
Asunto(s)
Esclerosis Múltiple/metabolismo , Vaina de Mielina/metabolismo , Receptores de Interleucina-8B/metabolismo , Remielinización/fisiología , Animales , Axones/metabolismo , Axones/patología , Diferenciación Celular/fisiología , Línea Celular , Sistema Nervioso Central/patología , Enfermedades Desmielinizantes/virología , Modelos Animales de Enfermedad , Humanos , Ratones , Ratones Transgénicos , Esclerosis Múltiple/patología , Virus de la Hepatitis Murina/metabolismo , Neuroglía/metabolismo , Neuroglía/patología , Oligodendroglía/metabolismo , Receptores de Interleucina-8B/genética , Transducción de Señal , Células Madre , Tamoxifeno/farmacologíaRESUMEN
MicroRNA (miRNA) regulation of gene expression is becoming an increasingly recognized mechanism by which host immune responses are governed following microbial infection. miRNAs are short, non-coding RNAs that repress translation of target genes, and have been implicated in a number of activities that modulate host immune responses, including the regulation of immune cell proliferation, survival, expansion, differentiation, migration, polarization, and effector function. This review highlights several examples in which mammalian-encoded miR-155 influences immune responses following viral infection of the CNS.
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Encefalitis , MicroARNs/genética , Mielitis , Virosis/complicaciones , Animales , Encefalitis/etiología , Encefalitis/inmunología , Encefalitis/virología , Regulación Viral de la Expresión Génica/inmunología , Humanos , MicroARNs/metabolismo , Mielitis/etiología , Mielitis/inmunología , Mielitis/virologíaRESUMEN
BACKGROUND: MicroRNAs (miRNAs) are noncoding RNAs that modulate cellular gene expression, primarily at the post-transcriptional level. We sought to examine the functional role of miR-155 in a model of viral-induced neuroinflammation. METHODS: Acute encephalomyelitis and immune-mediated demyelination were induced by intracranial injection with the neurotropic JHM strain of mouse hepatitis virus (JHMV) into C57BL/6 miR-155 (+/+) wildtype (WT) mice or miR-155 (-/-) mice. Morbidity and mortality, viral load and immune cell accumulation in the CNS, and spinal cord demyelination were assessed at defined points post-infection. T cells harvested from infected mice were used to examine cytolytic activity, cytokine activity, and expression of certain chemokine receptors. To determine the impact of miR-155 on trafficking, T cells from infected WT or miR-155 (-/-) mice were adoptively transferred into RAG1 (-/-) mice, and T cell accumulation into the CNS was assessed using flow cytometry. Statistical significance was determined using the Mantel-Cox log-rank test or Student's T tests. RESULTS: Compared to WT mice, JHMV-infected miR-155 (-/-) mice developed exacerbated disease concomitant with increased morbidity/mortality and an inability to control viral replication within the CNS. In corroboration with increased susceptibility to disease, miR-155 (-/-) mice had diminished CD8(+) T cell responses in terms of numbers, cytolytic activity, IFN-γ secretion, and homing to the CNS that corresponded with reduced expression of the chemokine receptor CXCR3. Both IFN-γ secretion and trafficking were impaired in miR-155 (-/-) , virus-specific CD4(+) T cells; however, expression of the chemokine homing receptors analyzed on CD4(+) cells was not affected. Except for very early during infection, there were not significant differences in macrophage infiltration into the CNS between WT and miR-155 (-/-) JHMV-infected mice, and the severity of demyelination was similar at 14 days p.i. between WT and miR-155 (-/-) JHMV-infected mice. CONCLUSIONS: These findings support a novel role for miR-155 in host defense in a model of viral-induced encephalomyelitis. Specifically, miR-155 enhances antiviral T cell responses including cytokine secretion, cytolytic activity, and homing to the CNS in response to viral infection. Further, miR-155 can play either a host-protective or host-damaging role during neuroinflammation depending on the disease trigger.
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Movimiento Celular/genética , Infecciones por Coronavirus/complicaciones , MicroARNs/metabolismo , Enfermedades del Sistema Nervioso/patología , Enfermedades del Sistema Nervioso/terapia , Linfocitos T/fisiología , Traslado Adoptivo/métodos , Animales , Sistema Nervioso Central/patología , Sistema Nervioso Central/fisiopatología , Sistema Nervioso Central/virología , Infecciones por Coronavirus/inmunología , Citocinas/metabolismo , Modelos Animales de Enfermedad , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , MicroARNs/genética , Enfermedades del Sistema Nervioso/etiología , Factores de TiempoRESUMEN
Respiratory syncytial virus (RSV) is a negative-strand RNA virus that is an important cause of bronchiolitis and pneumonia. We investigated the effect of RSV infection on the expression patterns of cellular proteins involved in regulating mRNA translation and degradation, and found that a processing-body protein involved in mRNA degradation, decapping protein 1a (DCP1), was phosphorylated rapidly following infection. UV-inactivated and sucrose-purified RSV were sufficient to mediate DCP1 phosphorylation, indicating that it occurs as a consequence of an early event in RSV infection. Analysis using kinase inhibitors showed that RSV-induced DCP1 phosphorylation occurred through the ERK1/2 pathway. The DCP1 phosphorylation sites were limited to serine 315, serine 319, and threonine 321. Overexpression of wt DCP1 led to a decrease in RSV-induced IL-8 production, but this effect was abrogated in cells overexpressing phosphorylation-deficient DCP1 mutants. These results suggest that DCP1 phosphorylation modulates the host chemokine response to RSV infection.
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Endorribonucleasas/metabolismo , Interleucina-8/genética , Infecciones por Virus Sincitial Respiratorio/metabolismo , Virus Sincitial Respiratorio Humano/fisiología , Transactivadores/metabolismo , Secuencias de Aminoácidos , Endorribonucleasas/química , Endorribonucleasas/genética , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Células Hep G2 , Humanos , Interleucina-8/metabolismo , Fosforilación , Infecciones por Virus Sincitial Respiratorio/genética , Infecciones por Virus Sincitial Respiratorio/virología , Transactivadores/química , Transactivadores/genéticaRESUMEN
Clinicians and patients often confuse drug names that sound alike. We conducted auditory perception experiments in the United States to assess the impact of similarity, familiarity, background noise and other factors on clinicians' (physicians, family pharmacists, nurses) and laypersons' ability to identify spoken drug names. We found that accuracy increased significantly as the signal-to-noise (S/N) ratio increased, as subjective familiarity with the name increased and as the national prescribing frequency of the name increased. For clinicians only, similarity to other drug names reduced identification accuracy, especially when the neighboring names were frequently prescribed. When one name was substituted for another, the substituted name was almost always a more frequently prescribed drug. Objectively measurable properties of drug names can be used to predict confusability. The magnitude of the noise and familiarity effects suggests that they may be important targets for intervention. We conclude that the ability of clinicians and lay people to identify spoken drug names is influenced by signal-to-noise ratio, subjective familiarity, prescribing frequency, and the similarity neighborhoods of drug names.
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Errores de Medicación , Habla , Adulto , Factores de Edad , Percepción Auditiva , Femenino , Audición , Humanos , Masculino , Enfermeras y Enfermeros , Pacientes , Farmacéuticos , Médicos , Medicamentos bajo Prescripción , Factores de Riesgo , Factores SexualesRESUMEN
OBJECTIVES: To determine the incidence rate of inpatient stroke-induced aphasia in Ontario, Canada, and to examine the demographic and clinical characteristics for stroke patients with and without aphasia. DESIGN: Age- and sex-specific incidence rates for aphasia in Ontario were calculated using the Ontario Stroke Audit. In addition, data collected from the Registry of the Canadian Stroke Network (RCSN) were used to determine the demographic and clinical characteristics for stroke patients with and without aphasia. SETTING: All hospitals and regional stroke centers in Ontario, Canada. PARTICIPANTS: The Ontario Stroke Audit is a representative weighted sample of more than 3000 stroke inpatients admitted to emergency departments in all hospitals in Ontario within the 2004/2005 fiscal year. RCSN data included a cohort of more than 15,000 consecutive patients presenting with stroke at 12 regional stroke centers in Ontario from 2003 to 2007. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Presence of aphasic symptoms on admission to hospital and at discharge, age and sex, stroke type and severity, severity of disability, services received in hospital, length of stay, and discharge destination. RESULTS: Thirty-five percent (1131/3207) of adult patients admitted with a diagnosis of stroke in the province of Ontario during the 2004 to 2005 Ontario Stroke Audit had symptoms of aphasia at the time of discharge. This amounts to an incidence rate of 60 per 100,000 persons per year. Risk of aphasia increased significantly with age. In comparison with nonaphasic stroke patients, patients with aphasia were older, presented with more severe strokes on admission, had more severe disability, and were more frequently discharged to long-term care and/or rehabilitation (unadjusted results). Adjusting for stroke severity, age, sex, comorbidity, and stroke subtype, the presence of aphasia was found to be an independent predictor of longer hospital stays, increased use of rehabilitation services, and higher rates of thrombolytic therapy. CONCLUSIONS: A significant number of people with stroke experience aphasia, with advancing age associated with a higher risk. The profile and patterns for stroke patients with aphasia differed significantly from those who did not experience aphasia as a residual disability after stroke, particularly in relation to service usage. Given the personal and system cost associated with aphasia, best practices in the area of stroke should include recommendations on how to best serve this population throughout the clinical pathway.
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Afasia/epidemiología , Afasia/rehabilitación , Hospitalización/estadística & datos numéricos , Rehabilitación de Accidente Cerebrovascular , Accidente Cerebrovascular/epidemiología , Adulto , Distribución por Edad , Anciano , Anciano de 80 o más Años , Afasia/diagnóstico , Estudios de Casos y Controles , Femenino , Humanos , Incidencia , Masculino , Auditoría Médica , Persona de Mediana Edad , Ontario/epidemiología , Sistema de Registros , Distribución por Sexo , Accidente Cerebrovascular/complicaciones , Adulto JovenRESUMEN
Bacterial vaginosis (BV) is a complex vaginal infection most commonly associated with women of child-bearing age. Risk factors for BV are numerous. There are currently multiple clinical and laboratory tests for diagnosis of BV, including the most commonly used diagnostic methods: Amsel's criteria or Nugent's Gram stain scale. The mainstay of BV therapy is metronidazole, but tinidazole as well as a few other agents have also been used successfully. Tinidazole is the second nitroimidazole antiprotozoal agent and a structural derivative of metronidazole. With a favorable pharmacokinetic profile and reduced side effects, tinidazole is an alternative agent for BV treatment. There are minimal head-to-head comparative data to establish tinidazole's superiority to metronidazole or other therapeutic agents. Available data suggest tinidazole has a role in special populations particularly for refractory or relapsing BV.