Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 4.323
Filtrar
1.
Immunogenetics ; 72(1-2): 57-76, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31761977

RESUMO

The Immune Epitope Database and Analysis Resource (IEDB) contains information related to antibodies and T cells across an expansive scope of research fields (infectious diseases, allergy, autoimmunity, and transplantation). Capture and representation of the data to reflect growing scientific standards and techniques have required continual refinement of our rigorous curation and query and reporting processes beginning with the automated classification of over 28 million PubMed abstracts, and resulting in easily searchable data from over 20,000 published manuscripts. Data related to MHC binding and elution, nonpeptidics, natural processing, receptors, and 3D structure is first captured through manual curation and subsequently maintained through recuration to reflect evolving scientific standards. Upon promotion to the free, public database, users can query and export records of specific relevance via the online web portal which undergoes iterative development to best enable efficient data access. In parallel, the companion Analysis Resource site hosts a variety of tools that assist in the bioinformatic analyses of epitopes and related structures, which can be applied to IEDB-derived and independent datasets alike. Available tools are classified into two categories: analysis and prediction. Analysis tools include epitope clustering, sequence conservancy, and more, while prediction tools cover T and B cell epitope binding, immunogenicity, and TCR/BCR structures. In addition to these tools, benchmarking servers which allow for unbiased performance comparison are also offered. In order to expand and support the user-base of both the database and Analysis Resource, the research team actively engages in community outreach through publication of ongoing work, conference attendance and presentations, hosting of user workshops, and the provision of online help. This review provides a description of the IEDB database infrastructure, curation and recuration processes, query and reporting capabilities, the Analysis Resource, and our Community Outreach efforts, including assessment of the impact of the IEDB across the research community.


Assuntos
Bases de Dados de Proteínas , Epitopos/imunologia , Proteínas/imunologia , Animais , Humanos , Fatores de Tempo
2.
Anal Bioanal Chem ; 411(29): 7725-7735, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31760445

RESUMO

The rapid and simultaneous detection of DNA and protein biomarkers is necessary to detect the outbreak of a disease or to monitor a disease. For example, cardiovascular diseases are a major cause of adult mortality worldwide. We have developed a rapidly adaptable platform to assess biomarkers using a microfluidic technology. Our model mimics autoantibodies against three proteins, C-reactive protein (CRP), brain natriuretic peptide (BNP), and low-density lipoprotein (LDL). Cell-free mitochondrial DNA (cfmDNA) and DNA controls are detected via fluorescence probes. The biomarkers are covalently bound on the surface of size- (11-15 µm) and dual-color encoded microbeads and immobilized as planar layer in a microfluidic chip flow cell. Binding events of target molecules were analyzed by fluorescence measurements with a fully automatized fluorescence microscope (end-point and real-time) developed in house. The model system was optimized for buffers and immobilization strategies of the microbeads to enable the simultaneous detection of protein and DNA biomarkers. All prime target molecules (anti-CRP, anti-BNP, anti-LDL, cfmDNA) and the controls were successfully detected both in independent reactions and simultaneously. In addition, the biomarkers could also be detected in spiked human serum in a similar way as in the optimized buffer system. The detection limit specified by the manufacturer is reduced by at least a factor of five for each biomarker as a result of the antibody detection and kinetic experiments indicate that nearly 50 % of the fluorescence intensity is achieved within 7 min. For rapid data inspection, we have developed the open source software digilogger, which can be applied for data evaluation and visualization. Graphical abstract.


Assuntos
Doenças Cardiovasculares/metabolismo , Ácidos Nucleicos Livres/análise , Dispositivos Lab-On-A-Chip , Proteínas/análise , Autoanticorpos/análise , Biomarcadores/análise , Corantes Fluorescentes/química , Humanos , Limite de Detecção , Microesferas , Proteínas/imunologia
3.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 41(5): 714-718, 2019 Oct 30.
Artigo em Chinês | MEDLINE | ID: mdl-31699206

RESUMO

Leucine-rich glioma-inactivated protein 1(LGI1)antibody-associated encephalitis is an autoimmune brain disease mainly seen in mid-aged and elderly people.Its main clinical manifestations include abnormal mental behaviors,facial-arm dystonia,hyponatremia,and hypokalemia.Immunotherapy with gamma globulin and/or hormone is effective.Two patients with LGT1 antibody-associated encephalitis were diagnosed in our center between January 2018 and October 2018,with typical clinical findings.The disease was cursed after immunoglobulin and hormone treatments.


Assuntos
Autoanticorpos/imunologia , Encefalite/diagnóstico , Proteínas/imunologia , Encefalite/terapia , Humanos
4.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 41(3): 344-350, 2019 Jun 30.
Artigo em Chinês | MEDLINE | ID: mdl-31282328

RESUMO

Objective To explore the clinical characteristics of autoimmune disease with dual seropositive antibodies of leucine-rich glioma inactivated 1(LGI1)and contactin-associated protein 2(Caspr2).Methods The clinical data of seven patients with dual seropositive LGI1 and Caspr2 antibodies who were admitted to the Neurology Department of Peking Union Medical College Hospital from July 2014 to December 2017 were retrospectively analyzed.Results Central,peripheral and autonomic nervous systems were all involved in the seven cases;100%(7/7)presented with insomnia,myokymia,neuropahic pain and hyperhydrosis;71%(5/7)showed memory decline or psychiatric and behavioral symptoms;57%(4/7)had urinary hesitation or constipation;and 43%(3/7)had seizure.Electromyography showed 100%(6/6) of the patients had prolonged afterdischarges following normal M waves and/or abnormal spontaneous firing.Electroencephalography revealed slow waves or basic rhythm slowing in 71%(5/7)of patients.Electrocardiography showed sinus tachycardia,axis deviation,and prolonged QT intervals in 71%(5/7)of patients.One patient died from arrhythmia before immunotherapy.One died from pulmonary infection after immunotherapy.Improvement with immunotherapy was documented in the other five cases.No relapse was noted during the 1-2-year follow-up.Conclusions Autoimmune disease with dual seropositive antibodies of LGI1 and Caspr2 can diffusely affect the central,peripheral,and autonomic nervous systems.The possibility of this disease should be considered in patients with acute and subacute onset of neuropsychiatric symptoms,especially in patients with accompanying insomnia,myokymia,and hyperhydrosis.


Assuntos
Autoanticorpos/sangue , Doenças Autoimunes/imunologia , Proteínas de Membrana/imunologia , Proteínas do Tecido Nervoso/imunologia , Proteínas/imunologia , Humanos , Estudos Retrospectivos
5.
PLoS Pathog ; 15(6): e1007880, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31211814

RESUMO

The largest ever recorded epidemic of the Chikungunya virus (CHIKV) broke out in 2004 and affected four continents. Acute symptomatic infections are typically associated with the onset of fever and often debilitating polyarthralgia/polyarthritis. In this study, a systems biology approach was adopted to analyze the blood transcriptomes of adults acutely infected with the CHIKV. Gene signatures that were associated with viral RNA levels and the onset of symptoms were identified. Among these genes, the putative role of the Eukaryotic Initiation Factor (eIF) family genes and apolipoprotein B mRNA editing catalytic polypeptide-like (APOBEC3A) in the CHIKV replication process were displayed. We further compared these signatures with signatures induced by the Dengue virus infection and rheumatoid arthritis. Finally, we demonstrated that the CHIKV in vitro infection of murine bone marrow-derived macrophages induced IL-1 beta production in a mechanism that is significantly dependent on the inflammasome NLRP3 activation. The observations provided valuable insights into virus-host interactions during the acute phase and can be instrumental in the investigation of new and effective therapeutic interventions.


Assuntos
Artrite/imunologia , Febre de Chikungunya/imunologia , Vírus Chikungunya/fisiologia , Citidina Desaminase/imunologia , Proteínas/imunologia , Replicação Viral/imunologia , Adulto , Animais , Artrite/patologia , Artrite/virologia , Febre de Chikungunya/patologia , Vírus da Dengue/imunologia , Vírus da Dengue/patogenicidade , Feminino , Febre/imunologia , Febre/patologia , Febre/virologia , Seguimentos , Humanos , Interleucina-1beta/imunologia , Camundongos , Proteína 3 que Contém Domínio de Pirina da Família NLR/imunologia
6.
Cancer Treat Rev ; 77: 35-43, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31207478

RESUMO

Immunotherapeutic strategies have revolutionised cancer therapy in recent years, bringing meaningful improvements in outcomes for patients with previously intractable conditions. These successes have, however, been largely limited to certain types of liquid tumours and a small subset of solid tumours that are known to be particularly immunogenic. Broadening these advances across the majority of tumour indications, which are characterised by an immune-excluded, immune-deserted or immune-suppressed ('cold') phenotype, will require alternative approaches that are able to specifically address this unique biological environment. Several newer therapeutic modalities, including adoptive cell therapy and T cell redirecting bispecific molecules, are considered to hold particular promise and are being investigated in early phase clinical trials across various solid tumour indications. ImmTAC molecules are a novel class of T cell redirecting bispecific biologics that exploit TCR-based targeting of tumour cells; providing potent and highly specific access to the vast landscape of intracellular targets. The first of these reagents to reach the clinic, tebentafusp (IMCgp100), has generated demonstrable clinical efficacy in an immunologically cold solid tumour with a high unmet need. Here, we highlight the key elements of the ImmTAC platform that make it ideally positioned to overcome the cold tumour microenvironment in an off-the-shelf format.


Assuntos
Imunoterapia/métodos , Neoplasias/imunologia , Neoplasias/terapia , Receptores de Antígenos de Linfócitos T/imunologia , Linfócitos T/imunologia , Antígenos de Neoplasias/imunologia , Produtos Biológicos/administração & dosagem , Humanos , Imunoterapia Adotiva/métodos , Proteínas/imunologia , Anticorpos de Cadeia Única/imunologia , Antígeno gp100 de Melanoma/imunologia
7.
Int J Mol Med ; 44(2): 363-374, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31198979

RESUMO

Chronic, low­grade inflammation associated with obesity and diabetes result from the infiltration of adipose and vascular tissue by immune cells and contributes to cardiovascular complications. Despite an incomplete understanding of the mechanistic underpinnings of immune cell differentiation and inflammation, O­GlcNAcylation, the addition of O­linked N­acetylglucosamine (O­GlcNAc) to cytoplasmic, nuclear and mitochondrial proteins by the two cycling enzymes, the O­linked N­acetylglucosamine transferase (OGT) and the O­GlcNAcase (OGA), may contribute to fine­tune immunity and inflammation in both physiological and pathological conditions. Early studies have indicated that O­GlcNAcylation of proteins play a pro­inflammatory role in diabetes and insulin resistance, whereas subsequent studies have demonstrated that this post­translational modification could also be protective against acute injuries. These studies suggest that diverse types of insults result in dynamic changes to O­GlcNAcylation patterns, which fluctuate with cellular metabolism to promote or inhibit inflammation. In this review, the current understanding of O­GlcNAcylation and its adaptive modulation in immune and inflammatory responses is summarized.


Assuntos
Acetilglucosamina/imunologia , Imunidade , Inflamação/imunologia , Animais , Humanos , Inflamação/patologia , N-Acetilglucosaminiltransferases/imunologia , Proteínas/imunologia , beta-N-Acetil-Hexosaminidases/imunologia
8.
Mol Cell ; 75(2): 394-407.e5, 2019 07 25.
Artigo em Inglês | MEDLINE | ID: mdl-31227230

RESUMO

The structural diversity of glycans on cells-the glycome-is vast and complex to decipher. Glycan arrays display oligosaccharides and are used to report glycan hapten binding epitopes. Glycan arrays are limited resources and present saccharides without the context of other glycans and glycoconjugates. We used maps of glycosylation pathways to generate a library of isogenic HEK293 cells with combinatorially engineered glycosylation capacities designed to display and dissect the genetic, biosynthetic, and structural basis for glycan binding in a natural context. The cell-based glycan array is self-renewable and reports glycosyltransferase genes required (or blocking) for interactions through logical sequential biosynthetic steps, which is predictive of structural glycan features involved and provides instructions for synthesis, recombinant production, and genetic dissection strategies. Broad utility of the cell-based glycan array is demonstrated, and we uncover higher order binding of microbial adhesins to clustered patches of O-glycans organized by their presentation on proteins.


Assuntos
Engenharia Genética , Redes e Vias Metabólicas/genética , Polissacarídeos/química , Proteínas/genética , Epitopos/genética , Epitopos/imunologia , Glicosilação , Glicosiltransferases/genética , Células HEK293 , Humanos , Oligossacarídeos/genética , Polissacarídeos/classificação , Polissacarídeos/genética , Polissacarídeos/imunologia , Proteínas/imunologia
9.
Biomed Res Int ; 2019: 1315257, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31111043

RESUMO

Prenatal environmental exposures are considered to contribute to the development of allergic sensitization by epigenetic mechanisms. The role of histone acetylation in the placenta has not been examined yet. We hypothesized that placental histone acetylation at the promoter regions of allergy-related immune regulatory genes is associated with the development of sensitization to allergens in the child. Histones H3 and H4 acetylation at the promoter regions of 6 selected allergy-related immune regulatory genes was assessed by a chromatin immunoprecipitation assay in 173 term placentas collected in the prospective birth-cohort ALADDIN. The development of IgE sensitization to allergens in the children was followed from 6 months up to 5 years of age. We discovered significant associations of histone acetylation levels with decreased risk of allergic sensitization in 3 genes. Decreased risk of sensitization to food allergens was associated with higher H3 acetylation levels in placentas at the IFNG and SH2B3 genes, and for H4 acetylation in HDAC4. Higher HDAC4 H4 acetylation levels were also associated with a decreased risk of sensitization to aeroallergens. In conclusion, our results suggest that acetylation of histones in placenta has a potential to predict the development of sensitization to allergens in children.


Assuntos
Hipersensibilidade Alimentar/genética , Histona Desacetilases/genética , Interferon gama/genética , Proteínas/genética , Proteínas Repressoras/genética , Acetilação , Alérgenos/efeitos adversos , Pré-Escolar , Cromatina/genética , Epigênese Genética/genética , Epigênese Genética/imunologia , Feminino , Hipersensibilidade Alimentar/imunologia , Hipersensibilidade Alimentar/patologia , Histona Desacetilases/imunologia , Histonas/genética , Histonas/imunologia , Humanos , Imunoglobulina E/genética , Imunoglobulina E/imunologia , Lactente , Recém-Nascido , Interferon gama/imunologia , Peptídeos e Proteínas de Sinalização Intracelular , Masculino , Placenta/imunologia , Placenta/patologia , Gravidez , Regiões Promotoras Genéticas/genética , Proteínas/imunologia , Proteínas Repressoras/imunologia
10.
Nature ; 569(7757): 509-513, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-31068699

RESUMO

A universal gain-of-function approach for selective and temporal control of protein activity in living systems is crucial to understanding dynamic cellular processes. Here we report development of a computationally aided and genetically encoded proximal decaging (hereafter, CAGE-prox) strategy that enables time-resolved activation of a broad range of proteins in living cells and mice. Temporal blockage of protein activity was computationally designed and realized by genetic incorporation of a photo-caged amino acid in proximity to the functional site of the protein, which can be rapidly removed upon decaging, resulting in protein re-activation. We demonstrate the wide applicability of our method on diverse protein families, which enabled orthogonal tuning of cell signalling and immune responses, temporal profiling of proteolytic substrates upon caspase activation as well as the development of protein-based pro-drug therapy. We envision that CAGE-prox will open opportunities for the gain-of-function study of proteins and dynamic biological processes with high precision and temporal resolution.


Assuntos
Sobrevivência Celular , Proteínas/metabolismo , Animais , Caspase 3/genética , Caspase 3/metabolismo , Linhagem Celular , Ativação Enzimática , Mutação com Ganho de Função , Humanos , Masculino , Camundongos , Neoplasias/tratamento farmacológico , Fosfotransferases/metabolismo , Pró-Fármacos/metabolismo , Pró-Fármacos/uso terapêutico , Proteínas/genética , Proteínas/imunologia , Proteínas/uso terapêutico , Proteólise , Proteômica , Transdução de Sinais , Fatores de Tempo
11.
Food Funct ; 10(6): 3477-3490, 2019 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-31140472

RESUMO

Rhubarb, a well-used herbal and dietary supplement, has been widely used as a laxative in many countries. The dietary supplement rhubarb may reveal differential hepatotoxicity responses in normal and diseased subjects; however, its underlying mechanism is unclear. By using a network pharmacology approach, we found that the components contained in rhubarb had associations with a liver disease-related protein network that could be enriched into two subnetworks: a pro-inflammatory protein network associated with liver inflammation and an anti-inflammatory protein network related to liver fibrosis. In addition, macrophages were found to have an association with these subnetworks. Herein, the differential toxicity responses of rhubarb in normal and diseased rats were illustrated by in vivo pharmacology experiments. Rhubarb induced liver injury in normal rats with dose-dependent increases in the pro-inflammatory response; in contrast, it failed to induce hepatotoxic effects in a liver fibrosis rat model and was accompanied by an increase in anti-inflammatory protein expression. Further study showed elevation of high mobility group box-1 (HMGB1) in the sera and liver tissues and remarkable pro-inflammatory activation of Kupffer cells in liver tissue; these phenomena were associated with the hepatotoxic effect of rhubarb and could be blocked by inhibiting either HMGB1 or Kupffer cells through glycyrrhizin or GdCl3, respectively. Interestingly, we also observed attenuated pro-inflammatory activation of Kupffer cells in a liver fibrosis rat model together with a non-hepatotoxic response to rhubarb. These results suggest that the divergent immune states in normal and diseased subjects may contribute to the differential toxicity responses to rhubarb.


Assuntos
Doença Hepática Induzida por Substâncias e Drogas/etiologia , Medicamentos de Ervas Chinesas/administração & dosagem , Fígado/efeitos dos fármacos , Rheum/química , Animais , Doença Hepática Induzida por Substâncias e Drogas/genética , Doença Hepática Induzida por Substâncias e Drogas/imunologia , Doença Hepática Induzida por Substâncias e Drogas/patologia , Medicamentos de Ervas Chinesas/efeitos adversos , Medicamentos de Ervas Chinesas/química , Humanos , Fígado/imunologia , Fígado/patologia , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Masculino , Proteínas/genética , Proteínas/imunologia , Ratos , Ratos Sprague-Dawley , Rheum/efeitos adversos
12.
Mol Ther ; 27(7): 1262-1274, 2019 07 03.
Artigo em Inglês | MEDLINE | ID: mdl-31043341

RESUMO

Chimeric antigen receptor (CAR) T cell therapies have demonstrated impressive initial response rates in hematologic malignancies. However, relapse rates are significant, and robust efficacies in other indications, such as solid tumors, will likely require novel therapeutic strategies and CAR designs. To that end, we sought to develop simple, highly selective targeting domains (D domains) that could be incorporated into complex, multifunctional therapeutics. Herein, we describe the identification and characterization of D domains specific for CD123, a therapeutic target for hematologic malignancies, including acute myelogenous leukemia (AML). CARs comprised of these D domains mediate potent T cell activation and cytolysis of CD123-expressing target cells and induce complete durable remission in two AML xenograft models. We describe a strategy of engineering less immunogenic D domains through the identification and removal of putative T cell epitopes and investigate the binding kinetics and affinity requirements of the resultant D domain CARs. Finally, we extended the utility of D domains by generating functional, bi-specific CARs comprised of a CD123-specific D domain and a CD19-specific scFv. The properties of D domains suggest that this class of targeting domain may facilitate the development of multi-functional CARs where conventional, scFv-based designs may be suboptimal.


Assuntos
Antineoplásicos/uso terapêutico , Imunoterapia Adotiva/métodos , Subunidade alfa de Receptor de Interleucina-3/imunologia , Domínios Proteicos/imunologia , Proteínas/uso terapêutico , Receptores de Antígenos Quiméricos/uso terapêutico , Linfócitos T/imunologia , Animais , Antígenos CD19/imunologia , Antineoplásicos/imunologia , Epitopos de Linfócito T/imunologia , Células HEK293 , Humanos , Células K562 , Leucemia Mieloide Aguda/terapia , Ativação Linfocitária/imunologia , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Ligação Proteica/imunologia , Proteínas/imunologia , Receptores de Antígenos Quiméricos/imunologia , Anticorpos de Cadeia Única/imunologia , Transfecção , Ensaios Antitumorais Modelo de Xenoenxerto
13.
Vet Microbiol ; 232: 42-49, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31030843

RESUMO

Aspergillosis is a fungal infection caused by Aspergillus molds that can affect both humans and animals. Despite advances in diagnostics and therapy, medical management of this disease remains difficult. Expansion of the basic knowledge regarding its pathophysiology in animals is critical to aid in the identification of new biomarkers of infection for diagnosis and therapeutic targets. For such a purpose, proteomics can be used by addressing protein changes during various disease processes. In the present study, a mass spectrometry analysis based on isobaric tagging for relative and absolute quantitation (iTRAQ®) was applied for direct identification and relative quantitation of proteins in blood collected from 32 Aspergillus-diseased common bottlenose dolphins (Tursiops truncatus, 32 samples) in comparison with blood from 55 other dolphins (55 samples from 41 clinically-normal controls and from 14 cetaceans with miscellaneous non-Aspergillus inflammation diseases) and ten convalescent dolphins (28 samples). Sixty-six and 40 proteins were found to be ≥2.0-fold over- and underrepresented versus miscellaneous non-Aspergillus inflammatory dolphins, respectively, and most were confirmed vs. clinically-normal controls and convalescents. Many proteins which play a role in the adaptive immune response were identified, including MHC proteins and others involved in catalytic activity like the NADPH-ubiquinone oxido-reductases. Overall, iTRAQ® appears to be a convenient proteomic tool greatly suited for exploratory ex vivo studies focusing on pathophysiology. This technique should be considered as a preliminary step before validation of new diagnostic markers.


Assuntos
Imunidade Adaptativa , Aspergilose/fisiopatologia , Aspergilose/veterinária , Golfinho Nariz-de-Garrafa/imunologia , Proteínas/imunologia , Animais , Aspergillus , Biomarcadores/sangue , Golfinho Nariz-de-Garrafa/microbiologia , Feminino , Masculino , Espectrometria de Massas , Proteômica
14.
J Toxicol Sci ; 44(4): 283-297, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30944281

RESUMO

We previously developed a test for detecting naturally occurring protein-induced skin sensitization based on the markers and criteria of the human cell-line activation test (h-CLAT) and showed that the h-CLAT was useful for assessing the allergenic potency of proteins. However, test proteins were contaminated with varying amounts of lipopolysaccharide (LPS), which might have contributed to the stimulation of CD86 and CD54 expression. In this study, we developed a method to exclude the effects of LPS in the assessment of skin sensitization by naturally occurring proteins. We tested two inhibitors [the caspase-1 inhibitor acetyl-Tyr-Val-Ala-Asp-chloromethylketone (Ac-YVAD-cmk; hereafter referred to as YVAD), which can mitigate the LPS-induced increases in CD54 expression, and polymyxin B (PMB), which suppresses the effect of LPS by binding to its lipid moiety (i.e., the toxic component of LPS)]. After a 24 hr exposure, YVAD and PMB reduced LPS-induced CD86 and CD54 expression. In particular, the effect of PMB was dependent upon pre-incubation time and temperature, with the most potent effect observed following pre-incubation at 37°C for 24 hr. Moreover, only pre-incubation with cell-culture medium (CCM) at 37°C for 24 hr showed an inhibitory effect similar to that of PMB, with this result possibly caused by components of CCM binding to LPS. Similar effects were observed in the presence of ovalbumin (with 1070 EU/mg LPS) and ovomucoid, and lysozyme (with 2.82 and 0.234 EU/mg LPS, respectively) in CCM. These results indicated that PMB and CCM effectively eliminated the effects of LPS during assessment of protein allergenicity, thereby allowing a more accurate evaluation of the potential of proteins to induce skin sensitization.


Assuntos
Clorometilcetonas de Aminoácidos/farmacologia , Hipersensibilidade/etiologia , Hipersensibilidade/imunologia , Lipopolissacarídeos/antagonistas & inibidores , Lipopolissacarídeos/imunologia , Polimixina B/farmacologia , Proteínas/efeitos adversos , Proteínas/imunologia , Testes Cutâneos/métodos , Pele/imunologia , Clorometilcetonas de Aminoácidos/administração & dosagem , Clorometilcetonas de Aminoácidos/metabolismo , Antígeno B7-2/genética , Antígeno B7-2/imunologia , Antígeno B7-2/metabolismo , Meios de Cultura , Expressão Gênica/efeitos dos fármacos , Humanos , Imunização , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/imunologia , Molécula 1 de Adesão Intercelular/metabolismo , Lipopolissacarídeos/efeitos adversos , Lipopolissacarídeos/metabolismo , Muramidase , Ovalbumina , Ovomucina , Polimixina B/administração & dosagem , Polimixina B/metabolismo , Ligação Proteica , Células THP-1 , Temperatura Ambiente , Fatores de Tempo
15.
RNA ; 25(6): 713-726, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30894411

RESUMO

Viral and cellular double-stranded RNA (dsRNA) is recognized by cytosolic innate immune sensors, including RIG-I-like receptors. Some cytoplasmic dsRNA is commonly present in cells, and one source is mitochondrial dsRNA, which results from bidirectional transcription of mitochondrial DNA (mtDNA). Here we demonstrate that Trp53 mutant mouse embryonic fibroblasts contain immune-stimulating endogenous dsRNA of mitochondrial origin. We show that the immune response induced by this dsRNA is mediated via RIG-I-like receptors and leads to the expression of type I interferon and proinflammatory cytokine genes. The mitochondrial dsRNA is cleaved by RNase L, which cleaves all cellular RNA including mitochondrial mRNAs, increasing activation of RIG-I-like receptors. When mitochondrial transcription is interrupted there is a subsequent decrease in this immune-stimulatory dsRNA. Our results reveal that the role of p53 in innate immunity is even more versatile and complex than previously anticipated. Our study, therefore, sheds new light on the role of endogenous RNA in diseases featuring aberrant immune responses.


Assuntos
Adenosina Desaminase/genética , Proteína DEAD-box 58/genética , Imunidade Inata/genética , RNA de Cadeia Dupla/genética , RNA Mitocondrial/genética , Proteína Supressora de Tumor p53/genética , Adenosina Desaminase/deficiência , Adenosina Desaminase/imunologia , Animais , Proteínas de Transporte/genética , Proteínas de Transporte/imunologia , Proteína DEAD-box 58/imunologia , Embrião de Mamíferos , Endorribonucleases/genética , Endorribonucleases/imunologia , Fibroblastos/citologia , Fibroblastos/imunologia , Fator Regulador 7 de Interferon/genética , Fator Regulador 7 de Interferon/imunologia , Helicase IFIH1 Induzida por Interferon/genética , Helicase IFIH1 Induzida por Interferon/imunologia , Peptídeos e Proteínas de Sinalização Intracelular , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Proteínas/genética , Proteínas/imunologia , RNA de Cadeia Dupla/imunologia , RNA Mitocondrial/imunologia , Proteínas de Ligação a RNA , Transcrição Genética , Transfecção , Proteína Supressora de Tumor p53/deficiência , Proteína Supressora de Tumor p53/imunologia
17.
BMC Neurol ; 19(1): 19, 2019 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-30732585

RESUMO

BACKGROUND: Anti leucine-rich glioma inactivated 1 (LGI1) encephalitis is a rare autoimmune encephalitis (AE), characterized by acute or subacute cognitive impairment, faciobrachial dystonic seizures, psychiatric disturbances and hyponatremia. Antibody-LGI 1 autoimmune encephalitis (anti-LGI1 AE) has increasingly been recognized as a primary autoimmune disorder with favorable prognosis and response to treatment. CASE PRESENTATION: Herein, we reported a male patient presenting as rapidly progressive dementia and hyponatremia. He had antibodies targeting LGI1 both in the cerebrospinal fluid and serum, which demonstrated the diagnosis of typical anti-LGI1 AE. The scores of Mini-Mental State Examination and Montreal Cognitive Assessment were 19/30 and 15/30, respectively. Cranial magnetic resonance images indicated hyperintensities in bilateral hippocampus. The findings of brain arterial spin labeling and Fluorine-18-fluorodeoxyglucose positron emission tomography showed no abnormal perfusion/metabolism. After the combined treatment of intravenous immunoglobulin and glucocorticoid, the patient's clinical symptoms improved obviously. CONCLUSIONS: This case raises the awareness that a rapid progressive dementia with predominant memory deficits could be induced by immunoreactions against LGI1. The better recognition will be great importance for the early diagnosis, essential treatment, even a better prognosis.


Assuntos
Demência/etiologia , Hiponatremia/etiologia , Encefalite Límbica/complicações , Encefalite Límbica/diagnóstico , Proteínas/imunologia , Autoanticorpos/sangue , Autoantígenos/imunologia , Encéfalo/patologia , Humanos , Encefalite Límbica/imunologia , Masculino , Pessoa de Meia-Idade
18.
PLoS Pathog ; 15(2): e1007609, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30779786

RESUMO

Kaposi's sarcoma-associated herpesvirus (KSHV) is causally associated with Kaposi's sarcoma, primary effusion lymphoma (PEL) and multicentric Castleman's disease. The IFIT family of proteins inhibits replication of some viruses, but their effects on KSHV lytic replication was unknown. Here we show that KSHV lytic replication induces IFIT expression in epithelial cells. Depletion of IFIT1, IFIT2 and IFIT3 (IFITs) increased infectious KSHV virion production 25-32-fold compared to that in control cells. KSHV lytic gene expression was upregulated broadly with preferential activation of several genes involved in lytic viral replication. Intracellular KSHV genome numbers were also increased by IFIT knockdown, consistent with inhibition of KSHV DNA replication by IFITs. RNA seq demonstrated that IFIT depletion also led to downregulation of IFN ß and several interferon-stimulated genes (ISGs), especially OAS proteins. OAS down-regulation led to decreased RNase L activity and slightly increased total RNA yield. IFIT immunoprecipitation also showed that IFIT1 bound to viral mRNAs and cellular capped mRNAs but not to uncapped RNA or trimethylated RNAs, suggesting that IFIT1 may also inhibit viral mRNA expression through direct binding. In summary, IFIT inhibits KSHV lytic replication through positively regulating the IFN ß and OAS RNase L pathway to degrade RNA in addition to possibly directly targeting viral mRNAs.


Assuntos
Proteínas de Transporte/imunologia , Proteínas de Transporte/metabolismo , Herpesvirus Humano 8/metabolismo , Replicação do DNA/genética , Regulação para Baixo , Células Epiteliais , Regulação Viral da Expressão Gênica/genética , Células HEK293 , Herpesvirus Humano 8/patogenicidade , Humanos , Imunidade Inata/fisiologia , Interferon beta/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/imunologia , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Proteínas/imunologia , Proteínas/metabolismo , RNA , RNA Mensageiro , Ativação Viral , Latência Viral/genética , Replicação Viral/genética
19.
J Dent Res ; 98(5): 541-548, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30779877

RESUMO

Dental enamel malformations, or amelogenesis imperfecta (AI), can be isolated or syndromic. To improve the prospects of making a successful diagnosis by genetic testing, it is important that the full range of genes and mutations that cause AI be determined. Defects in WDR72 (WD repeat-containing protein 72; OMIM *613214) cause AI, type IIA3 (OMIM #613211), which follows an autosomal recessive pattern of inheritance. The defective enamel is normal in thickness, severely hypomineralized, orange-brown stained, and susceptible to attrition. We identified 6 families with biallelic WDR72 mutations by whole exome sequence analyses that perfectly segregated with the enamel phenotype. The novel mutations included 3 stop-gains [NM_182758.2: c.377G>A/p.(Trp126*), c.1801C>T/p.(Arg601*), c.2350A>T/p.(Arg784*)], a missense mutation [c.1265G>T/p.(Gly422Val)], and a 62,138-base pair deletion (NG_017034.2: g.35441_97578del62138) that removed WDR72 coding exons 3 through 13. A previously reported WDR72 frameshift was also observed [c.1467_1468delAT/p.(Val491Aspfs*8)]. Three of the affected patients showed decreased serum pH, consistent with a diagnosis of renal tubular acidosis. Percentiles of stature and body weight varied among 8 affected individuals but did not show a consistent trend. These studies support that WDR72 mutations cause a syndromic form of AI and improve our ability to diagnose AI caused by WDR72 defects.


Assuntos
Acidose , Amelogênese Imperfeita , Proteínas/imunologia , Acidose/genética , Amelogênese Imperfeita/genética , Humanos , Mutação , Linhagem
20.
Clin Rev Allergy Immunol ; 57(2): 261-271, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30734159

RESUMO

Food protein-induced enterocolitis syndrome (FPIES) is a non-IgE-mediated food allergy that has been well-characterized clinically, yet it is still poorly understood. Acute FPIES is characterized by vomiting 1-4 h and/or diarrhea within 24 h after ingestion of a culprit food. Chronic FPIES is the result of chronic exposure to an offending food that can result in chronic watery diarrhea, intermittent vomiting, and failure to thrive. FPIES typically presents in infancy and self-resolves by school age in most patients. Adult-onset FPIES is rare, but it has been reported. Cow's milk and soy are the most common triggering foods in infants in the US, and as solids are introduced in the diet, FPIES reactions to grains (rice, oat) increase in prevalence. Variability in common trigger foods exists depending on the geographical origin-for example, fish is a frequent trigger in Spanish and Italian patients. Heavy reliance on a detailed history is required for the diagnosis as physical exam findings, laboratory tests, and/or imaging studies are suggestive and not specific for FPIES. Oral food challenges remain the gold standard for confirming diagnosis, and the challenge protocol may be for an individual depending on risk of reaction, prior reaction severity, and positive-specific IgE status. The recent development of diagnostic criteria in 2017 will serve to increase recognition of the disorder and allow for early implementation of management strategies. Acute management during reactions includes IV hydration, anti-emetics, and IV corticosteroids. Reaction prevention strategies include strict food avoidance until the physician deems a food reintroduction challenge clinically appropriate. Future efforts in FPIES research should be aimed at elucidating the underlying disease mechanisms and possible treatment targets.


Assuntos
Enterocolite/epidemiologia , Enterocolite/imunologia , Hipersensibilidade Alimentar/epidemiologia , Hipersensibilidade Alimentar/imunologia , Proteínas/imunologia , Adolescente , Corticosteroides/uso terapêutico , Adulto , Alérgenos/imunologia , Antieméticos/uso terapêutico , Criança , Pré-Escolar , Diagnóstico Diferencial , Dieta , Enterocolite/diagnóstico , Enterocolite/terapia , Feminino , Hidratação , Humanos , Imunoglobulina E/análise , Incidência , Lactente , Masculino , Ondansetron/uso terapêutico , Testes do Emplastro
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA