Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 272
Filtrar
1.
Allergol. immunopatol ; 49(1): 95-100, ene.-feb. 2021. tab
Artigo em Inglês | IBECS | ID: ibc-199231

RESUMO

INTRODUCTION AND OBJECTIVES: The purpose of this study was to evaluate patients diagnosed with 22q11.2 deletion syndrome and determine the clues directing to diagnosis and evaluation of immunological findings for excellent management of the disease. MATERIAL AND METHODS: Thirty-three pediatric patients with 22q11.2 deletion syndrome diag­nosed between 1998 and 2019 at Pediatric Immunology Division of Ege University Faculty of Medicine and SBU Izmir Dr Behcet Uz Children's Education and Research Hospital were evaluated. RESULTS: This study includes the largest case series reported from Turkey. Congenital car­diac anomalies were the most common pathology associated with the syndrome (90.9%). Hypocalcemic symptoms were observed in 13 patients (40%). Twenty-two of the 33 (66.6%) patients were diagnosed before two years of age. Autoimmune diseases, dysmorphic facial findings, recurrent infections, growth retardation, and speech impairment were other clues for diagnosis in older patients. Clinical spectrum and immunological abnormalities of this syn­drome are quite variable. All T-cell subset counts were less than 5th percentile below median by age in one patient (3%) and 10 patients had normal all T-cell subset counts (30.3%). Overall, 69.6% of the patients had normal IgG, IgA, and IgM levels and two patients had panhypogam­maglobulinemia. Recurrent infections were revealed in 75.7% of the patients during follow-up. CONCLUSIONS: Presence of cardiac anomaly is more helpful in the diagnosis, especially under two years of age. Patients with immunologically high or standard risk did not show any differ­ence in terms of numbers and severity of infections and autoimmunity


No disponible


Assuntos
Humanos , Masculino , Feminino , Pré-Escolar , Síndrome da Deleção 22q11/diagnóstico , Síndromes de Imunodeficiência/epidemiologia , Síndromes de Imunodeficiência/imunologia , Transtornos Cromossômicos/epidemiologia , Cromossomos Humanos Par 22 , Síndrome da Deleção 22q11/imunologia , Doenças do Sistema Imunitário/diagnóstico , Doenças do Sistema Imunitário/imunologia , Testes Imunológicos , Técnicas Imunológicas/métodos , Doenças Autoimunes/diagnóstico , Doenças Autoimunes/imunologia
2.
Methods Mol Biol ; 2209: 217-234, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33201472

RESUMO

Stress granules are dynamic structures that assemble in response to various forms of stress, such as heat shock or oxidative stress, among others. We had previously shown that the lysine deacetylase HDAC6 is required for the formation of stress granules, but the substrate important for this function was not known. We recently found that the RNA helicase DDX3X is a novel HDAC6 substrate, which is critical for the formation of stress granules. Through a series of detailed experiments, we showed that, upon stress, DDX3X becomes acetylated in an intrinsically disordered region; this alters its propensity to undergo phase separation and inhibits growth of the stress granules. HDAC6, by deacetylating DDX3X, allows maturation of the stress granules. This work identified acetylation of an RNA helicase as a critical regulator of the stress response. Here, we present methods to analyze the acetylation of DDX3X; these methods can be easily adapted to study the acetylation of other helicases, or other proteins.


Assuntos
RNA Helicases DEAD-box/metabolismo , Resposta ao Choque Térmico , Técnicas Imunológicas/métodos , Estresse Oxidativo , Acetilação , Linhagem Celular , Humanos , Processamento de Proteína Pós-Traducional
4.
Allergol. immunopatol ; 48(3): 259-264, mayo-jun. 2020. tab
Artigo em Inglês | IBECS | ID: ibc-192028

RESUMO

The clinical history is of importance in the investigation of allergic diseases but does have limitations. Many allergic conditions will be over-diagnosed if anamnesis alone is used for diagnostic criteria. Serum total immunoglobulin E (TIgE) quantification, as well as panels containing allergens prevalent in the studied population, may serve as screening tests and facilitate the diagnosis of allergic disease or its exclusion. We assessed the positivity of two versions of these tests, Phadiatop Europe® (PhEU) and Phadiatop Infant® (PhInf), as well as total IgE (TigE) values in patients with a medical diagnosis of allergic disease and non-allergic individuals. METHODS: A cross-sectional study performed in eleven Brazilian pediatric allergy centers with patients divided into groups according to the primary condition and a group of assessed control subjects. They were submitted to TIgE measurement and screening tests (PhEu and PhInf). RESULTS: TIgE mean serum levels were significantly higher among allergic patients, especially those with asthma/rhinitis or atopic dermatitis. The positivity of the screening tests, considering the total population, was 63.8% for PhEU and 72.6% for PhInf. These increased when we evaluated only the allergic subjects. The concordance index of the two tests was Kappa = 0.7 and higher among those of greater age. CONCLUSIONS: In the assessed population, there were significantly higher levels among those with positive screening tests and PhInf showed better performance in the identification of sensitized individuals, regardless of age. This is the first study to evaluate Phadiatop and Phadiatop Infant in the same population


No disponible


Assuntos
Humanos , Lactente , Pré-Escolar , Criança , Adolescente , Testes Hematológicos/métodos , Hipersensibilidade/diagnóstico , Imunoglobulina E/sangue , Técnicas Imunológicas/métodos , Hipersensibilidade/imunologia , Brasil , Estudos Transversais , Alérgenos/imunologia , Dermatite Atópica/imunologia , Asma/imunologia , Rinite Alérgica/imunologia , Sensibilidade e Especificidade
5.
J Aquat Anim Health ; 31(4): 328-348, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31634422

RESUMO

Macrophage aggregates (MAs) are focal accumulations of pigmented macrophages in the spleen and other tissues of fish. A central role of MAs is the clearance and destruction of degenerating cells and recycling of some cellular components. Macrophage aggregates also respond to chemical contaminants and infectious agents and may play a role in the adaptive immune response. Tissue damage or physiological stress can result in increased MA accumulation. As a result, MAs may be sensitive biomarkers of environmental stress in fish. Abundance of MAs in tissues has been reported in a variety of ways-most commonly as density, mean size, and relative area-but the utility of these estimates has not been compared. In this study, four different types of splenic MA abundance estimates (abundance score, density, relative area, and total volume) were compared in two fish populations (Striped Bass Morone saxatilis and White Perch M. americana) with a wide range in ages. Stereological estimates of total volume indicated an increase in MA abundance with spleen volume, which generally corresponded to fish age, and with splenic infections (mycobacteria or trematode parasites). Abundance scores were generally limited in the ability to detect changes in MA abundance by these factors, whereas density estimates were greatly influenced by changes in spleen volume. In some instances, densities declined while the total volume of MAs and spleen volume increased. Experimentally induced acute stress resulted in a decrease in spleen volume and an increase in MA density, although the total volume of MAs remained unchanged. Relative area estimates accounted for the size and number of MAs but not for changes in organ volume. Total volume is an absolute measure of MA abundance irrespective of changes in organ volume or patterns of accumulation and may provide an improved means of quantifying MAs in the spleens of fish.


Assuntos
Bass/imunologia , Técnicas Imunológicas/veterinária , Macrófagos/fisiologia , Baço/imunologia , Estresse Fisiológico/imunologia , Animais , Feminino , Doenças dos Peixes/imunologia , Técnicas Imunológicas/instrumentação , Técnicas Imunológicas/métodos , Masculino , Esplenopatias/imunologia , Esplenopatias/veterinária
6.
J Vis Exp ; (150)2019 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-31475984

RESUMO

The purification of monoclonal antibodies (mAbs) is commonly achieved by Protein A affinity chromatography, which can account for up to 25% of the overall process costs. Alternative, cost-effective capture steps are therefore valuable for industrial-scale manufacturing, where large quantities of a single mAb are produced. Here we present a method for the immobilization of a DsRed-based epitope ligand to a cross-linked agarose resin allowing the selective capture of the HIV-neutralizing antibody 2F5 from crude plant extracts without using Protein A. The linear epitope ELDKWA was first genetically fused to the fluorescent protein DsRed and the fusion protein was expressed in transgenic tobacco (Nicotiana tabacum) plants before purification by immobilized metal-ion affinity chromatography. Furthermore, a method based on activated cross-linked agarose was optimized for high ligand density, efficient coupling and low costs. The pH and buffer composition and the soluble ligand concentration were the most important parameters during the coupling procedure, which was improved using a design-of-experiments approach. The resulting affinity resin was tested for its ability to selectively bind the target mAb in a crude plant extract and the elution buffer was optimized for high mAb recovery, product activity and affinity resin stability. The method can easily be adapted to other antibodies with linear epitopes. The new resins allow gentler elution conditions than Protein A and could also reduce the costs of an initial capture step for mAb production.


Assuntos
Anticorpos Monoclonais/química , Anticorpos Amplamente Neutralizantes/química , Cromatografia de Afinidade/métodos , Anticorpos Anti-HIV/química , Técnicas Imunológicas/métodos , Sefarose/química , Epitopos/química , Ligantes , Extratos Vegetais , Proteínas de Plantas , Plantas Geneticamente Modificadas , Proteína Estafilocócica A , Tabaco/genética , Tabaco/metabolismo
7.
Clin Lab ; 65(9)2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31532090

RESUMO

BACKGROUND: Multiplex bead assays, also known as addressable laser bead immunoassays (ALBIA) or Luminex® technology, have provided an alternative to enzyme-linked immunoassay, which is still the most widely utilized routine immunoassay for detection of specific autoantibodies. Our laboratory adopted the ALBIA technology early into its routine service. METHODS: We report the performance and utility of measurement of three different autoantibody types tested using the FIDIS (BMD, Marne La Vallee, France) ALBIA system. The analytes discussed are thyroid antibodies (thyroglobulin [TG], thyroid peroxidase [TPO]), anti-neutrophil cytoplasmic antibodies (ANCA), and ribonucleo-protein (RNP) antibodies. RESULTS: In single antibody analysis, TPO antibody testing was superior to TG antibody in identifying patients with Graves' disease and Hashimoto's thyroiditis. However, testing only TPO antibody would result in missing 8.6% of Graves' and 11.9% of Hashimoto's thyroiditis patients, hence demonstrating an advantage for the multiplex TG plus TPO assay. With respect to ANCA, the FIDIS ALBIA produced an overall similar level of performance to our comparator method, the Phadia fluorescent enzyme linked immunoassay. Sensitivity of the ALBIA for RNP antibodies was low in comparison to countercurrent immunoelectrophoresis, but performance was improved by altering the cutoff value for the assay. CONCLUSIONS: ALBIA technology has many potential advantages in the routine laboratory, but as with any new assay, evaluation must be thorough and ongoing to ensure satisfactory clinical performance is obtained. Both false positive and false negative results have been reported in ALBIA studies. It may be necessary to re-evaluate assay performance and cutoff and consider further clinical correlation.


Assuntos
Anticorpos Anticitoplasma de Neutrófilos/análise , Autoanticorpos/análise , Imunoensaio/métodos , Iodeto Peroxidase/imunologia , Ribonucleoproteínas/imunologia , Adulto , Anticorpos Anticitoplasma de Neutrófilos/imunologia , Anticorpos Antinucleares/análise , Anticorpos Antinucleares/imunologia , Autoanticorpos/imunologia , Testes Diagnósticos de Rotina/métodos , Feminino , Humanos , Técnicas Imunológicas/métodos , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos Testes
8.
Fish Shellfish Immunol ; 93: 832-840, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31425832

RESUMO

Stingrays skin secretions are largely studied due to the human envenoming medical relevance of the sting puncture that evolves to inflammatory events, including necrosis. Such toxic effects can be correlated to the biochemical composition of the sting mucus, according to the literature. Fish skin plays important biological roles, such as the control of the osmotic pressure gradient, protection against mechanical forces and microorganism infections. The mucus, on the other hand, is a rich and complex fluid, acting on swimming, nutrition and the innate immune system. The elasmobranch's epidermis is a tissue composed mainly by mucus secretory cells, and marine stingrays have already been described to present secretory glands spread throughout the body. Little is known about the biochemical composition of the stingray mucus, but recent studies have corroborated the importance of mucus in the envenomation process. Aiming to assess the mucus composition, a new non-invasive mucus collection method was developed that focused on peptides and proteins, and biological assays were performed to analyze the toxic and immune activities of the Hypanus americanus mucus. Pathophysiological characterization showed the presence of peptidases on the mucus, as well as the induction of edema and leukocyte recruitment in mice. The fractionated mucus improved phagocytosis on macrophages and showed antimicrobial activity against T. rubrumç. neoformans and C. albicans in vitro. The proteomic analyses showed the presence of immune-related proteins like actin, histones, hemoglobin, and ribosomal proteins. This protein pattern is similar to those reported for other fish mucus and stingray venoms. This is the first report depicting the Hypanus stingray mucus composition, highlighting its biochemical composition and importance for the stingray immune system and the possible role on the envenomation process.


Assuntos
Venenos de Peixe/química , Imunidade Inata , Técnicas Imunológicas/veterinária , Muco/química , Animais , Brasil , Feminino , Imunidade nas Mucosas , Técnicas Imunológicas/métodos , Muco/imunologia , Rajidae
9.
Bioanalysis ; 11(17): 1605-1617, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31208198

RESUMO

Biological drug products may elicit an antidrug antibody (ADA) response. The current widely used bridging ligand binding assay (LBA) is the gold standard for ADA assessments in drug development, which is a qualitative assay followed by a quasi-quantitative titer analysis but can be prone to interferences from biological matrices, drug targets and circulating drugs. We present our perspectives and findings in exploring a hybrid LBA/LC-MS as an orthogonal bioanalytical tool for clinical immunogenicity assessments. The hybrid LBA/LC-MS is a semiquantitative assay with acceptable specificity, drug tolerance and the capability of multiplexed detection of ADA isotypes. The assay results suggest this technology to be a promising and complementary bioanalytical tool that can provide informative immunogenicity data in drug development.


Assuntos
Técnicas Imunológicas/métodos , Espectrometria de Massas , Artefatos , Cromatografia Líquida , Humanos , Limite de Detecção , Controle Social Formal
10.
Methods Mol Biol ; 1988: 199-215, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31147942

RESUMO

The 51Cr-release assay described in the 1960s has been for decades the gold standard cytolytic assay but for safety reasons is no longer in use in many laboratories. Whereas other radioactive tests were later on described, they never ousted the 51Cr-release assay. More thorough understanding of CTL biology and killing pathways has more recently resulted in the design of reliable nonradioactive tests to analyze CD8+ T cell responses. Allowing for real-time evaluation of target cell viability, some of them are particularly attractive and are described here together with traditional assays.


Assuntos
Linfócitos T CD8-Positivos/imunologia , Técnicas Imunológicas/métodos , Animais , ELISPOT , Humanos , Interferon gama/metabolismo , Camundongos , Linfócitos T Citotóxicos/imunologia
11.
Methods Mol Biol ; 1988: 357-373, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31147952

RESUMO

Macroautophagy has recently emerged as an important catabolic process involved not only in innate immunity but also in adaptive immunity. Initially described to deliver intracellular antigens to MHC class II loading compartments, its molecular machinery has now also been described to impact the delivery of extracellular antigens to MHC class II loading compartments through the noncanonical use of the macroautophagy machinery during LC3-associated phagocytosis (LAP). Therefore, in pathological situations (viral or bacterial infections, tumorigenesis) the pathway might be involved in shaping CD4+ T cell responses.In this chapter we describe three basic experiments for the monitoring and manipulation of macroautophagic antigen processing toward MHC class II presentation through the canonical pathway. Firstly, we will discuss how to monitor autophagic flux and autophagosome fusion with MHC class II loading compartments. Secondly, we will show how to target proteins to autophagosomes in order to monitor macroautophagy dependent antigen processing via their enhanced presentation on MHC class II molecules to CD4+ T cells. And finally, we will describe how macroautophagy can be silenced in antigen presenting cells, like human monocyte-derived dendritic cells (DCs).


Assuntos
Apresentação do Antígeno/imunologia , Antígenos de Histocompatibilidade Classe II/imunologia , Técnicas Imunológicas/métodos , Macroautofagia , Células A549 , Antígenos de Neoplasias/metabolismo , Autofagossomos/metabolismo , Linfócitos T CD4-Positivos/imunologia , Células Clonais , Células Dendríticas/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Células HEK293 , Humanos , Interferon gama/metabolismo , Proteínas de Membrana/metabolismo , Monócitos/citologia
12.
Methods Mol Biol ; 1988: 439-453, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31147957

RESUMO

Natural killer T (NKT) cells are a subset of T lymphocytes that recognize a wide variety of lipid antigens presented by the atypical MHC class I molecule CD1d. NKT cells exhibit rapid activation after recognition of cognate antigens, secrete abundant amounts of T helper (Th) 1, Th2, and Th17 cytokines within hours of activation and shape the immune response through subsequent activation of dendritic, NK, T, and B cells. NKT cells therefore play central roles in antimicrobial and anticancer immunity and in the modulation of various autoimmune disorders. Consequently, recent research has focused on the discovery of microbial and self-antigens involved in NKT cell activation. In this chapter, we will discuss different strategies for studying antigen recognition by NKT cells including CD1d tetramer-based approaches and in vitro assays characterizing NKT cell activation in response to lipid antigen presentation. While Toll-like receptor (TLR) agonists and cytokines such as IL-12 are critical for NKT cell activation in vivo, particularly in the context of microbial infection, methods for detection of TLR- and cytokine-dependent NKT cell activation will not be discussed in this section.


Assuntos
Apresentação do Antígeno/imunologia , Técnicas Imunológicas/métodos , Células T Matadoras Naturais/imunologia , Animais , Anticorpos/metabolismo , Células Apresentadoras de Antígenos/metabolismo , Antígenos CD1d/metabolismo , Linhagem Celular , Genoma , Humanos , Lipídeos/química , Fígado/metabolismo , Camundongos , Perfusão , Coloração e Rotulagem
13.
Elife ; 82019 06 24.
Artigo em Inglês | MEDLINE | ID: mdl-31232687

RESUMO

Previously, we described a novel alternative to chromatin immunoprecipitation, CUT&RUN, in which unfixed permeabilized cells are incubated with antibody, followed by binding of a protein A-Micrococcal Nuclease (pA/MNase) fusion protein (Skene and Henikoff, 2017). Here we introduce three enhancements to CUT&RUN: A hybrid protein A-Protein G-MNase construct that expands antibody compatibility and simplifies purification, a modified digestion protocol that inhibits premature release of the nuclease-bound complex, and a calibration strategy based on carry-over of E. coli DNA introduced with the fusion protein. These new features, coupled with the previously described low-cost, high efficiency, high reproducibility and high-throughput capability of CUT&RUN make it the method of choice for routine epigenomic profiling.


Assuntos
Cromatina/metabolismo , Técnicas Imunológicas/métodos , Biologia Molecular/métodos , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Epigênese Genética , Nuclease do Micrococo/genética , Nuclease do Micrococo/metabolismo , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Proteína Estafilocócica A/genética , Proteína Estafilocócica A/metabolismo
14.
Methods Mol Biol ; 1966: 211-224, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31041750

RESUMO

Activation of signal transducer and activator of transcription 6 (STAT6) is a key signaling pathway in macrophage function, and is required for the so-called alternative (M2) activation of macrophages. Interleukin (IL)-4 and IL-13 are important M2 polarizing cytokines that act through STAT6 by inducing its phosphorylation and promoting transcription of STAT6-responsive genes. Inactivation of STAT6 signaling in macrophages has not been fully explored; however, a recent model suggests that inactivation of STAT6 signaling can occur via ubiquitination and proteasomal degradation. In this chapter, we describe a combination of techniques that can be used to study the activation/inactivation of STAT6 signaling in macrophages.


Assuntos
Técnicas Imunológicas/métodos , Ativação de Macrófagos , Macrófagos/metabolismo , Fator de Transcrição STAT6/metabolismo , Transdução de Sinais , Animais , Linhagem Celular , Interleucina-4/metabolismo , Macrófagos/imunologia , Fosforilação , Processamento de Proteína Pós-Traducional
15.
Sci Rep ; 9(1): 5970, 2019 04 12.
Artigo em Inglês | MEDLINE | ID: mdl-30979922

RESUMO

Cardiac troponin T (cTnT) is considered a clinical standard for its high specificity and sensitivity when diagnosing acute myocardial infarction; however, most studies on the electrical sensors of cardiac troponin biomarkers have focused on cTnI rather than cTnT. This study presents label-free, low-cost, transparent, and flexible aptamer-based immunosensors for the electrical detection of cTnT using reduced graphene oxide (rGO) sheets. GO was first deposited by AC dielectrophoresis between two predefined source and drain electrodes on a 3-aminopropyltriethoxylsilane-modified polyethylene terephthalate substrate. The GO was then reduced using hydrazine vapour without damaging the substrate, resulting in uniform, controlled, and stable deposition of rGO sheets, and demonstrating more stability than those directly deposited by dielectrophoresis. Amine-modified single-strand DNA aptamers against cTnT were immobilized onto the rGO channels. The relative resistance change of this sensor owing to the attachment of cTnT was quantified as the cTnT concentration decreased from 10 ng/mL to 1 pg/mL in phosphate buffered saline (PBS) and 10-fold diluted human serum in PBS, with the limits of detection being 1.2 pg/mL and 1.7 pg/mL, respectively, which is sufficiently sensitive for clinical applications. High-yield and rapid fabrication of the present rGO sensors will have significant influences on scaled-up fabrication of graphene-based sensors.


Assuntos
Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais , Técnicas Eletroquímicas , Grafite/química , Técnicas Imunológicas , Troponina T/análise , Biomarcadores/análise , Técnicas Biossensoriais/métodos , Doenças Cardiovasculares/diagnóstico , Técnicas Eletroquímicas/métodos , Eletrodos , Desenho de Equipamento , Humanos , Técnicas Imunológicas/métodos , Sensibilidade e Especificidade
16.
J Allergy Clin Immunol Pract ; 7(7): 2225-2229.e1, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31034997

RESUMO

BACKGROUND: Misdiagnosis of amoxicillin (clavulanic acid) (AX(/CL)) hypersensitivity has serious consequences. A drug challenge (DC) is the final diagnostic to affirm or infirm AX(/CL) hypersensitivity. However, uncertainties remain whether a prolonged drug challenge (pDC) should benefit the diagnosis of a nonimmediate AX(/CL) hypersensitivity. OBJECTIVE: To assess the added value of a standardized 7-day pDC in the diagnosis of nonimmediate or unclear penicillin hypersensitivity. METHODS: A total of 132 patients with a history of a nonimmediate hypersensitivity reaction or an unclear reaction to AX(/CL) or an undefined penicillin with a negative diagnostic workup including a single-day DC (DC) with AX(/CL) were selected. In all these patients, an additional pDC with AX(/CL) was planned. Thirteen patients started the pDC immediately after the DC. To ensure that hypersensitivity symptoms manifesting during the pDC course do not result from the DC, in the remaining 119 patients, the pDC was scheduled after a washout of 1 week. RESULTS: A total of 128 patients (12 without washout, 116 with washout) completed the pDC. Three patients reacted with a mild maculopapular exanthema. However, the value of a pDC was evidenced in only 1 patient who reacted during her pDC after an uneventful washout. In 2 patients pDC was cancelled because they reacted during the washout. CONCLUSIONS: A pDC is of limited added value to the diagnostic algorithms of nonimmediate hypersensitivity reaction or unclear hypersensitivity reactions to AX(/CL). In our hands, the traditionally recommended diagnostic algorithm that offers a 1-day DC as a final diagnostic in patients with negative workup for AX(/CL) is appropriate.


Assuntos
Combinação Amoxicilina e Clavulanato de Potássio/efeitos adversos , Amoxicilina/efeitos adversos , Antibacterianos/efeitos adversos , Hipersensibilidade a Drogas/diagnóstico , Hipersensibilidade Tardia/diagnóstico , Técnicas Imunológicas/métodos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Amoxicilina/administração & dosagem , Antibacterianos/administração & dosagem , Criança , Pré-Escolar , Hipersensibilidade a Drogas/etiologia , Feminino , Humanos , Hipersensibilidade Tardia/etiologia , Masculino , Pessoa de Meia-Idade , Penicilinas/efeitos adversos , Estudos Retrospectivos , Fatores de Tempo , Adulto Jovem
17.
Sci Rep ; 9(1): 4834, 2019 03 18.
Artigo em Inglês | MEDLINE | ID: mdl-30886198

RESUMO

The complexity of immune responses limits the usefulness of univariate methods in answering complex immunology questions. To demonstrate the utility of a multivariate approach, we employ such approach to compare T cells of African green monkeys (AGMs) and rhesus macaques (RMs). Among the most prominent distinguishing features we found were lower CD3 and higher CD28 surface expression in AGMs compared to RMs. After in vitro stimulation, a larger proportion of AGM T cells secreted cytokines, especially those producing more than one cytokine (i.e. multifunctional cells). To find out whether multifunctional responses associate with protection in other species, we compared T cells of cynomolgus macaques (CMs) infected with wild-type Simian Immunodeficiency Virus (SIV) to those of CMs infected (vaccinated) with a replication-defective virus. Wild-type SIV infection in macaques leads to simian Acquired Immunodeficiency Syndrome (AIDS), which does not happen in animals previously vaccinated with a replication-defective virus. Interestingly, after in vitro stimulation, multifunctional cells were more abundant among T cells of vaccinated CMs. Our results propose T-cell multifunctionality as a potentially useful marker of immunity, although additional verification is needed. Finally, we hope our multivariate model and its associated validation methods will inform future studies in the field of immunology.


Assuntos
Técnicas Imunológicas/métodos , Vacinas contra a SAIDS/imunologia , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Subpopulações de Linfócitos T/imunologia , Linfócitos T/imunologia , Animais , Antígenos CD28/metabolismo , Complexo CD3/metabolismo , Chlorocebus aethiops/imunologia , Chlorocebus aethiops/virologia , Citocinas/imunologia , Citocinas/metabolismo , Imunogenicidade da Vacina , Contagem de Linfócitos , Macaca mulatta/imunologia , Macaca mulatta/virologia , Vacinas contra a SAIDS/administração & dosagem , Síndrome de Imunodeficiência Adquirida dos Símios/sangue , Síndrome de Imunodeficiência Adquirida dos Símios/virologia , Vírus da Imunodeficiência Símia/genética , Vírus da Imunodeficiência Símia/imunologia , Vírus da Imunodeficiência Símia/patogenicidade , Especificidade da Espécie , Subpopulações de Linfócitos T/metabolismo , Linfócitos T/metabolismo , Replicação Viral/genética , Replicação Viral/imunologia
19.
Cell Mol Immunol ; 16(3): 242-249, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30796351

RESUMO

The immune system is composed of a complex hierarchy of cell types that protect the organism against disease and maintain homeostasis. Identifying heterogeneity of immune cells is the key to understanding the immune system. Advanced single-cell RNA sequencing (scRNA-seq) technologies are revolutionizing our ability to study immunology. By measuring transcriptomes at the single-cell level, scRNA-seq enables identification of cellular heterogeneity in far greater detail than conventional methods. In this review, we introduce the existing scRNA-seq technologies and present their strengths and weaknesses. We also discuss potential applications and future innovations of scRNA-seq in immunology.


Assuntos
Alergia e Imunologia/tendências , Técnicas Imunológicas/métodos , Análise de Sequência de RNA/métodos , Análise de Célula Única/métodos , Animais , Perfilação da Expressão Gênica , Humanos , Transcriptoma
20.
Physiol Biochem Zool ; 92(2): 177-188, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30699048

RESUMO

The application of next-generation sequencing (NGS) to the study of whole genomes and transcriptomes is becoming commonplace in molecular biology and molecular medicine. Over the past decade, these technologies have become more accessible to the broader biological community as the cost of NGS has decreased significantly and the availability of ready-to-use library preparation kits and user-friendly bioinformatic tools has increased. Indeed, these technologies are starting to be deployed in the study of ecological systems and immune function and have already yielded new discoveries and insights. However, despite the power of these techniques, they remain underutilized by the ecological immunology (ecoimmunology) community. This is unfortunate because there are several key challenges faced by ecological immunologists that these technologies are particularly well suited to address: (1) to measure immune function in biologically meaningful ways, (2) to describe the complex relationship between immunology and other aspects of physiology in an ecological context, and (3) to relate ecoimmunology to disease ecology. Each of these three challenges confronting the ecoimmunology community is critical to understanding ecoimmunology, and each has been stymied by a lack of sufficient data to effectively tackle the questions at hand. Deploying NGS methods to tackle these challenges promises to revolutionize the field of ecoimmunology by facilitating the unbiased, broad-scale, and efficient discovery of novel immune genes and mechanisms; the quantification of variation in immune systems; the characterization of relevant physiological pathways; and the description of microbial communities.


Assuntos
Ecologia/métodos , Sequenciamento de Nucleotídeos em Larga Escala , Técnicas Imunológicas/métodos , Animais , Invertebrados/genética , Invertebrados/imunologia , Vertebrados/genética , Vertebrados/imunologia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...