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[PMID]:29295972
[Au] Autor:Jabs F; Plum M; Laursen NS; Jensen RK; Mølgaard B; Miehe M; Mandolesi M; Rauber MM; Pfützner W; Jakob T; Möbs C; Andersen GR; Spillner E
[Ad] Endereço:Immunological Engineering, Department of Engineering, Aarhus University, 8000, Aarhus, Denmark.
[Ti] Título:Trapping IgE in a closed conformation by mimicking CD23 binding prevents and disrupts FcεRI interaction.
[So] Source:Nat Commun;9(1):7, 2018 01 02.
[Is] ISSN:2041-1723
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Anti-IgE therapeutics interfere with the ability of IgE to bind to its receptors on effector cells. Here we report the crystal structure of an anti-IgE single-domain antibody in complex with an IgE Fc fragment, revealing how the antibody inhibits interactions between IgE and the two receptors FcεRI and CD23. The epitope overlaps only slightly with the FcεRI-binding site but significantly with the CD23-binding site. Solution scattering studies of the IgE Fc reveal that antibody binding induces a half-bent conformation in between the well-known bent and extended IgE Fc conformations. The antibody acts as functional homolog of CD23 and induces a closed conformation of IgE Fc incompatible with FcεRI binding. Notably the antibody displaces IgE from both CD23 and FcεRI, and abrogates allergen-mediated basophil activation and facilitated allergen binding. The inhibitory mechanism might facilitate strategies for the future development of anti-IgE therapeutics for treatment of allergic diseases.
[Mh] Termos MeSH primário: Epitopos/química
Imunoglobulina E/química
Receptores de IgE/química
[Mh] Termos MeSH secundário: Anticorpos Anti-Idiotípicos/química
Anticorpos Anti-Idiotípicos/metabolismo
Sítios de Ligação
Cristalografia por Raios X
Epitopos/metabolismo
Seres Humanos
Imunoglobulina E/metabolismo
Fragmentos Fc das Imunoglobulinas/química
Fragmentos Fc das Imunoglobulinas/metabolismo
Modelos Moleculares
Ligação Proteica
Conformação Proteica
Receptores de IgE/metabolismo
Anticorpos de Domínio Único/química
Anticorpos de Domínio Único/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Antibodies, Anti-Idiotypic); 0 (Epitopes); 0 (Immunoglobulin Fc Fragments); 0 (Receptors, IgE); 0 (Single-Domain Antibodies); 0 (anti-IgE antibodies); 37341-29-0 (Immunoglobulin E)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180306
[Lr] Data última revisão:
180306
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180104
[St] Status:MEDLINE
[do] DOI:10.1038/s41467-017-02312-7


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[PMID]:29329570
[Au] Autor:Erdei A; Steiber Z; Molnar C; Berenyi E; Nagy EV
[Ad] Endereço:Division of Endocrinology, Department of Medicine, Faculty of Medicine, University of Debrecen, Nagyerdei krt 98, Debrecen, 4032, Hungary. erdeianm@gmail.com.
[Ti] Título:Exophthalmos in a young woman with no graves' disease - a case report of IgG4-related orbitopathy.
[So] Source:BMC Ophthalmol;18(1):5, 2018 Jan 12.
[Is] ISSN:1471-2415
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Immunoglobulin G4-related disease (IgG4-rd) is characterized by lymphoplasmacytic infiltration and tissue fibrosis. Orbital manifestations of IgG4-rd may include unilateral or bilateral proptosis, cicatricial extraocular muscle myopathy, orbital inflammation and pain which may mimic ophthalmic Graves' disease. CASE PRESENTATION: A 25-year-old woman has been referred to the endocrinology clinic, 4 months after delivery, with suspected Graves' orbitopathy. She has had bronchial asthma and recurrent skin rashes of unknown aetiology for the last 10 years and was treated for dacryoadenitis with steroid containing eye drops 5 years ago. During pregnancy she developed eyelid swelling. After delivery, eyelid redness and retrobulbar pain evolved. Proptosis was demonstrated by Hertel's exophthalmometry. Orbital magnetic resonance imaging showed enlarged lateral and superior rectus muscles in both orbits. Thyroid function tests were in the normal range and no thyroid stimulating hormone (TSH) receptor autoantibodies were present. The eye muscle involvement pattern raised suspicion, and the high IgG4 level with positive histology of the lacrimal gland confirmed the diagnosis of immunoglobulin G4-related orbitopathy. Rapid improvement was observed following oral methylprednisolone. CONCLUSIONS: IgG4-related orbitopathy may mimic Graves' orbitopathy. Euthyroid patients with no TSH receptor autoantibodies should be evaluated for immunoglobulin G4-related orbitopathy. Once IgG4-related orbitopathy is proven, other manifestations of IgG4-related disease have to be searched for; lifelong follow-up is warranted.
[Mh] Termos MeSH primário: Anticorpos Anti-Idiotípicos/imunologia
Autoanticorpos/imunologia
Doenças Autoimunes/complicações
Exoftalmia/etiologia
Músculos Oculomotores/diagnóstico por imagem
[Mh] Termos MeSH secundário: Adulto
Doenças Autoimunes/diagnóstico
Doenças Autoimunes/imunologia
Diagnóstico Diferencial
Exoftalmia/diagnóstico
Exoftalmia/imunologia
Feminino
Oftalmopatia de Graves
Seres Humanos
Imagem por Ressonância Magnética
Órbita
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antibodies, Anti-Idiotypic); 0 (Autoantibodies); 0 (anti-IgA)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180305
[Lr] Data última revisão:
180305
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180114
[St] Status:MEDLINE
[do] DOI:10.1186/s12886-018-0672-y


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[PMID]:29293334
[Au] Autor:Qiu Y; Li P; Dong S; Zhang X; Yang Q; Wang Y; Ge J; Hammock BD; Zhang C; Liu X
[Ad] Endereço:Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology,Key Laboratory of Control Technology and Standard for Agro-product Safety and Quality (Ministry of Agriculture), Institute of Food Safety and Nutrition, Jiangsu Academy of Agricultural
[Ti] Título:Phage-Mediated Competitive Chemiluminescent Immunoassay for Detecting Cry1Ab Toxin by Using an Anti-Idiotypic Camel Nanobody.
[So] Source:J Agric Food Chem;66(4):950-956, 2018 Jan 31.
[Is] ISSN:1520-5118
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Cry toxins have been widely used in genetically modified organisms for pest control, raising public concern regarding their effects on the natural environment and food safety. In this work, a phage-mediated competitive chemiluminescent immunoassay (c-CLIA) was developed for determination of Cry1Ab toxin using anti-idiotypic camel nanobodies. By extracting RNA from camels' peripheral blood lymphocytes, a naive phage-displayed nanobody library was established. Using anti-Cry1Ab toxin monoclonal antibodies (mAbs) against the library for anti-idiotypic antibody screening, four anti-idiotypic nanobodies were selected and confirmed to be specific for anti-Cry1Ab mAb binding. Thereafter, a c-CLIA was developed for detection of Cry1Ab toxin based on anti-idiotypic camel nanobodies and employed for sample testing. The results revealed a half-inhibition concentration of developed assay to be 42.68 ± 2.54 ng/mL, in the linear range of 10.49-307.1 ng/mL. The established method is highly specific for Cry1Ab recognition, with negligible cross-reactivity for other Cry toxins. For spiked cereal samples, the recoveries of Cry1Ab toxin ranged from 77.4% to 127%, with coefficient of variation of less than 9%. This study demonstrated that the competitive format based on phage-displayed anti-idiotypic nanobodies can provide an alternative strategy for Cry toxin detection.
[Mh] Termos MeSH primário: Anticorpos Anti-Idiotípicos
Proteínas de Bactérias/análise
Bacteriófagos
Camelus/imunologia
Endotoxinas/análise
Proteínas Hemolisinas/análise
Medições Luminescentes/métodos
Anticorpos de Domínio Único
[Mh] Termos MeSH secundário: Animais
Anticorpos Monoclonais
Proteínas de Bactérias/imunologia
Camelus/sangue
Grãos Comestíveis/química
Endotoxinas/imunologia
Contaminação de Alimentos/análise
Proteínas Hemolisinas/imunologia
Linfócitos/química
Biblioteca de Peptídeos
RNA/isolamento & purificação
Anticorpos de Domínio Único/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antibodies, Anti-Idiotypic); 0 (Antibodies, Monoclonal); 0 (Bacterial Proteins); 0 (Endotoxins); 0 (Hemolysin Proteins); 0 (Peptide Library); 0 (Single-Domain Antibodies); 0 (insecticidal crystal protein, Bacillus Thuringiensis); 63231-63-0 (RNA)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180212
[Lr] Data última revisão:
180212
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180103
[St] Status:MEDLINE
[do] DOI:10.1021/acs.jafc.7b04923


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[PMID]:29236386
[Au] Autor:Galkin OY; Besarab AB; Lutsenko TN
[Ti] Título:Characteristics of enzyme-linked immunosorbent assay for detection of IgG antibodies specific to Сhlamydia trachomatis heat shock protein (HSP-60)
[So] Source:Ukr Biochem J;89(1):22-30, 2017 Jan-Feb.
[Is] ISSN:2409-4943
[Cp] País de publicação:Ukraine
[La] Idioma:eng
[Ab] Resumo:The goal of this work was to study sensitivity and specificity of the developed ELISA set for the identification of IgG antibodies against Chlamydia trachomatis HSP-60 (using biotinylated tyramine-based signal amplification system). The study was conducted using a panel of characterized sera, as well as two reference ELISA sets of similar purpose. According to the results of ELISA informative value parameters, the ELISA we have developed showed the highest specificity and sensitivity parameters (no false negative or false positive results were registered). In 4 out of 15 intralaboratory panel serum samples initially identified as negative, anti-HSP-60 IgG-antibodies test result in reference ELISA sets upon dilution changed from negative to positive. The nature of titration curves of false negative sera and commercial monoclonal antibodies А57-В9 against C. trachomatis HSP-60 after incubation for 24 h was indicative of the presence of anti-idiotypic antibodies in these samples. Upon sera dilution, idiotypic-anti-idiotypic complexes dissociated, which caused the change of test result. High informative value of the developed ELISA set for identification of IgG antibodies against C. trachomatis HSP-60 has been proven. Anti-idiotypic antibodies possessing C. trachomatis anti-HSP-60 activity and being one of the causes of false negative results of the relevant ELISA-based tests have been identified in blood sera of individuals infected with chlamydial genitourinary infection agents.
[Mh] Termos MeSH primário: Anticorpos Antibacterianos/análise
Antígenos de Bactérias/sangue
Chaperonina 60/sangue
Infecções por Chlamydia/diagnóstico
Chlamydia trachomatis/imunologia
Ensaio de Imunoadsorção Enzimática/métodos
Imunoglobulina G/análise
[Mh] Termos MeSH secundário: Anticorpos Anti-Idiotípicos/química
Anticorpos Antibacterianos/sangue
Anticorpos Antibacterianos/imunologia
Complexo Antígeno-Anticorpo/química
Antígenos de Bactérias/imunologia
Biotinilação
Chaperonina 60/imunologia
Infecções por Chlamydia/sangue
Infecções por Chlamydia/imunologia
Infecções por Chlamydia/microbiologia
Chlamydia trachomatis/química
Ensaio de Imunoadsorção Enzimática/normas
Reações Falso-Negativas
Seres Humanos
Soros Imunes/química
Imunoglobulina G/sangue
Imunoglobulina G/imunologia
Sensibilidade e Especificidade
Tiramina/química
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antibodies, Anti-Idiotypic); 0 (Antibodies, Bacterial); 0 (Antigen-Antibody Complex); 0 (Antigens, Bacterial); 0 (Chaperonin 60); 0 (Immune Sera); 0 (Immunoglobulin G); X8ZC7V0OX3 (Tyramine)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180116
[Lr] Data última revisão:
180116
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171214
[St] Status:MEDLINE
[do] DOI:10.15407/ubj89.01.022


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[PMID]:29187931
[Au] Autor:Fall S; Dieng F; Diouf C; Djiba B; Ndao AC; Ndiaye FSD
[Ad] Endereço:Unité d'Hématologie Clinique de l'Hôpital Aristide Le Dantec, Dakar, Sénégal.
[Ti] Título:[Diagnostic and evolutionary profile of multiple myeloma in Senegal: monocentric study conducted from 2005 to 2016].
[Ti] Título:Profil diagnostique et évolutif du myélome multiple au Sénégal: étude monocentrique de 2005 à 2016..
[So] Source:Pan Afr Med J;27:262, 2017.
[Is] ISSN:1937-8688
[Cp] País de publicação:Uganda
[La] Idioma:fre
[Ab] Resumo:Introduction: Accessibility to innovative multiple myeloma therapies is limited in sub-Saharan Africa. This study aimed to describe the diagnostic and evolutionary features observed during treatment of our patients with myeloma. Methods: We conducted a retrospective, descriptive, analytical study (2005 - 2016) of patients with myeloma included in the study based on International Myeloma Working Group (IMWG) Criteria (2003,2014) at the Hopital Aristide Le Dantec (Senegal). Results: We collected data from 136 medical records (69 men, 67 women) of patients with an average age of 59 years ± 10.1 years, who were less than 65 years of age in 69.1% of cases. Tell-tale signs included bone pain (96.3%), renal failure (36.8%), infection (23.5%), pathological fracture (17.6%), spinal cord compression (16.9%) and malignant hypercalcaemia (16.2%). Isotopic antiglobulin test showed that anti-IgG could be detected in 61.3% of cases and Kappa in 65% of cases. Patients were classified stage III (59.4%) and I-II (40.6%)of the index staging system. The median survival of patients under conventional traitement (Méphalan-Prédnisone: 67.6%, innovative: 5.9%) was 20 months (1-78 months). Survival rates are better in the absence of neurological and infectious complications and for patients with score I-II of the index Staging System. Conclusion: In our study, multiple myeloma was frequently diagnosed before age 65, at advanced stage of tumor mass. Early detection and access to adequate therapies could improve overall survival.
[Mh] Termos MeSH primário: Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico
Acesso aos Serviços de Saúde
Mieloma Múltiplo/terapia
[Mh] Termos MeSH secundário: Idoso
Anticorpos Anti-Idiotípicos/imunologia
Feminino
Seres Humanos
Masculino
Melfalan/administração & dosagem
Meia-Idade
Mieloma Múltiplo/diagnóstico
Mieloma Múltiplo/patologia
Estadiamento de Neoplasias
Prednisona/administração & dosagem
Estudos Retrospectivos
Senegal
Taxa de Sobrevida
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antibodies, Anti-Idiotypic); 0 (anti-IgG); Q41OR9510P (Melphalan); VB0R961HZT (Prednisone)
[Em] Mês de entrada:1712
[Cu] Atualização por classe:171219
[Lr] Data última revisão:
171219
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171201
[St] Status:MEDLINE
[do] DOI:10.11604/pamj.2017.27.262.13164


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[PMID]:28690180
[Au] Autor:Wang J; Wang J; Zhang Z; Zhang X; Ru S; Dong Y
[Ad] Endereço:Marine Life Science College, Ocean University of China, Qingdao 266003, China.
[Ti] Título:Development of an immunosensor for quantifying zebrafish vitellogenin based on the Octet system.
[So] Source:Anal Biochem;533:60-65, 2017 Sep 15.
[Is] ISSN:1096-0309
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Vitellogenin (Vtg) is a sensitive biomarker for environmental estrogens. In this study, an immunosensor for quantifying zebrafish Vtg was developed using the Octet system. First, Protein A sensors were immobilized with purified anti-lipovitellin (Lv) antibody that demonstrated specificity to Vtg. Then, antibody-coated biosensors were immersed into zebrafish Lv standards and diluted samples. The Octet system measured and recorded kinetic parameters between antigens and captured antibody within 5 min. Sample Vtg concentrations were automatically calculated by interpolating relative binding rates observed with each sample and the immobilized anti-Lv antibody into the developed standard curve. The sensor arrays exhibited a wide linear range from 78 to 5000 ng/mL, and the inter-assay coefficient of variation was 0.66-1.97%. Furthermore, the performance of the immunosensor in detecting Vtg was evaluated by quantifying Vtg induction in juvenile zebrafish exposed to 17ß-estradiol (E ). Compared with conventional immunoassay techniques, the Vtg immunosensor developed based on the Octet system was much simpler and less time-consuming, allowing rapid Vtg quantification within 15 min. Moreover, Protein A sensors could be reused many times to ensure that the assays have high reproducibility. Therefore, we suggest that immunosensors based on the Octet system are an easily operated detection method for ecotoxicological research.
[Mh] Termos MeSH primário: Anticorpos Anti-Idiotípicos/isolamento & purificação
Técnicas Biossensoriais
Vitelogeninas/isolamento & purificação
[Mh] Termos MeSH secundário: Animais
Anticorpos Anti-Idiotípicos/imunologia
Proteínas do Ovo/imunologia
Estradiol/farmacologia
Proteína Estafilocócica A/química
Vitelogeninas/imunologia
Peixe-Zebra/imunologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antibodies, Anti-Idiotypic); 0 (Egg Proteins); 0 (Staphylococcal Protein A); 0 (Vitellogenins); 4TI98Z838E (Estradiol); 9088-43-1 (lipovitellin)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170804
[Lr] Data última revisão:
170804
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170711
[St] Status:MEDLINE


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[PMID]:28652400
[Au] Autor:Schroeder JT; Bieneman AP
[Ad] Endereço:Division of Allergy and Clinical Immunology, Department of Medicine, Johns Hopkins Asthma and Allergy Center, Johns Hopkins University School of Medicine, Baltimore, MD 21224 schray@jhmi.edu.
[Ti] Título:Activation of Human Basophils by A549 Lung Epithelial Cells Reveals a Novel IgE-Dependent Response Independent of Allergen.
[So] Source:J Immunol;199(3):855-865, 2017 Aug 01.
[Is] ISSN:1550-6606
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Evidence for epithelial cell (EC)-derived cytokines (e.g., thymic stromal lymphopoietin [TSLP]) activating human basophils remains controversial. We therefore hypothesize that ECs can directly activate basophils via cell-to-cell interaction. Basophils in medium alone or with IL-3 ± anti-IgE were coincubated with TSLP, IL-33, or IL-25. Analogous experiments cocultured basophils (1-72 h) directly with EC lines. Supernatants were tested for mediators and cytokines. Abs targeting receptors were tested for neutralizing effects. Lactic acid (pH 3.9) treatment combined with passive sensitization tested the role of IgE. Overall, IL-33 augmented IL-13 secretion from basophils cotreated with IL-3, with minimal effects on histamine and IL-4. Conversely, basophils (but not mast cells) released histamine and marked levels of IL-4/IL-13 (10-fold) when cocultured with A549 EC and IL-3, without exogenous allergen or IgE cross-linking stimuli. The inability to detect IL-33 or TSLP, or to neutralize their activity, suggested a unique mode of basophil activation by A549 EC. Half-maximal rates for histamine (4 h) and IL-4 (5 h) secretion were slower than observed with standard IgE-dependent activation. Ig stripping combined with passive sensitization ± omalizumab showed a dependency for basophil-bound IgE, substantiated by a requirement for cell-to-cell contact, aggregation, and FcεRI-dependent signaling. A yet unidentified IgE-binding lectin associated with A549 EC is implicated after discovering that LacNAc suppressed basophil activation in cocultures. These findings point to a lectin-dependent activation of basophil requiring IgE but independent of allergen or secreted cytokine. Pending further investigation, we predict this unique mode of activation is linked to inflammatory conditions whereby IgE-dependent activation of basophils occurs despite the absence of any known allergen.
[Mh] Termos MeSH primário: Alérgenos/imunologia
Basófilos/imunologia
Células Epiteliais/imunologia
Imunoglobulina E/imunologia
[Mh] Termos MeSH secundário: Células A549
Anticorpos Anti-Idiotípicos/farmacologia
Basófilos/efeitos dos fármacos
Basófilos/metabolismo
Comunicação Celular
Técnicas de Cocultura
Citocinas/farmacologia
Citocinas/secreção
Células Epiteliais/metabolismo
Liberação de Histamina
Seres Humanos
Imunoglobulina E/metabolismo
Interleucina-13/imunologia
Interleucina-13/secreção
Interleucina-17/farmacologia
Interleucina-3/imunologia
Interleucina-3/farmacologia
Interleucina-33/farmacologia
Interleucina-4/imunologia
Interleucina-4/secreção
Ácido Láctico/farmacologia
Lectinas/metabolismo
Mastócitos/metabolismo
Omalizumab/farmacologia
Receptores de IgE/imunologia
Receptores de IgE/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Allergens); 0 (Antibodies, Anti-Idiotypic); 0 (Cytokines); 0 (FcepsilonRI alpha-chain, human); 0 (IL25 protein, human); 0 (IL3 protein, human); 0 (IL33 protein, human); 0 (Interleukin-13); 0 (Interleukin-17); 0 (Interleukin-3); 0 (Interleukin-33); 0 (Lectins); 0 (Receptors, IgE); 0 (anti-IgE antibodies); 0 (interleukin-13, human); 0 (thymic stromal lymphopoietin); 207137-56-2 (Interleukin-4); 2P471X1Z11 (Omalizumab); 33X04XA5AT (Lactic Acid); 37341-29-0 (Immunoglobulin E)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171010
[Lr] Data última revisão:
171010
[Sb] Subgrupo de revista:AIM; IM
[Da] Data de entrada para processamento:170628
[St] Status:MEDLINE
[do] DOI:10.4049/jimmunol.1700055


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[PMID]:28647457
[Au] Autor:Vigl B; Salhat N; Parth M; Pankevych H; Mairhofer A; Bartl S; Smrzka OW
[Ad] Endereço:AFFiRiS AG, Karl-Farkas-Gasse 22, 1030 Vienna, Austria. Electronic address: benjamin.vigl@affiris.com.
[Ti] Título:Quantitative in vitro and in vivo models to assess human IgE B cell receptor crosslinking by IgE and EMPD IgE targeting antibodies.
[So] Source:J Immunol Methods;449:28-36, 2017 Oct.
[Is] ISSN:1872-7905
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Targeting plasma IgE by therapeutic mABs like Omalizumab (Xolair ) is current clinical practice for severe allergic conditions or other IgE related diseases like chronic urticaria. As an alternative to soluble IgE targeting, IgE supply can be lowered by targeting the Extracellular Membrane Proximal Domain (EMPD) of the IgE B cell receptor (BCR) present on IgE switched B cells. This ultimately leads to apoptosis of these cells upon IgE BCR crosslinking. Since tools to selectively assess the efficacy of IgE BCR crosslinking by IgE targeting antibodies are limited, a readily quantifiable cell model was developed that allows to specifically address IgE BCR crosslinking activity in vitro. The new cell model allowed for a direct quantitative comparison of anti-EMPD IgE therapeutic prototype antibody 47H4 with anti-IgE(Ce3) directed therapeutic antibody Omalizumab and with a newly selected anti-human EMPD IgE monoclonal antibody, designated mAB 15cl12. Furthermore, a complementing mouse model was developed that allows for in vivo validation of antibodies addressing human EMPD IgE. It carries a targetable humanized EMPD IgE sequence that has been introduced by seamless genomic replacement of the endogenous EMPD encoding sequence. The model allowed to directly compare IgE lowering activity of two anti-human EMPD IgE therapeutic antibodies in vivo. Our tools provide the means for quantitative assessment of IgE BCR crosslinking activity which is increasingly gaining attention with respect to forthcoming second generation anti-IgE clinical candidates such as Ligelizumab or other clinical candidates featuring additional effector functions such as IgE BCR crosslinking activity.
[Mh] Termos MeSH primário: Anticorpos Anti-Idiotípicos/imunologia
Imunoglobulina E/química
Imunoglobulina E/imunologia
Receptores de Antígenos de Linfócitos B/química
Receptores de Antígenos de Linfócitos B/imunologia
[Mh] Termos MeSH secundário: Animais
Antialérgicos/química
Antialérgicos/metabolismo
Anticorpos Anti-Idiotípicos/química
Anticorpos Anti-Idiotípicos/metabolismo
Reagentes para Ligações Cruzadas
Seres Humanos
Imunoglobulina E/biossíntese
Imunoglobulina E/metabolismo
Camundongos
Omalizumab/química
Omalizumab/metabolismo
Receptores de Antígenos de Linfócitos B/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Anti-Allergic Agents); 0 (Antibodies, Anti-Idiotypic); 0 (Cross-Linking Reagents); 0 (Receptors, Antigen, B-Cell); 0 (anti-IgE antibodies); 2P471X1Z11 (Omalizumab); 37341-29-0 (Immunoglobulin E)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171018
[Lr] Data última revisão:
171018
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170626
[St] Status:MEDLINE


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[PMID]:28625864
[Au] Autor:Kavita U; Duo J; Crawford SM; Liu R; Valcin J; Gleason C; Dong H; Gadkari S; Dodge RW; Pillutla RC; DeSilva BS
[Ad] Endereço:Analytical and Bioanalytical Operations, Bristol-Myers Squibb, Princeton, NJ 08543, United States. Electronic address: uma.kavita@bms.com.
[Ti] Título:A systematic study of the effect of low pH acid treatment on anti-drug antibodies specific for a domain antibody therapeutic: Impact on drug tolerance, assay sensitivity and post-validation method assessment of ADA in clinical serum samples.
[So] Source:J Immunol Methods;448:91-104, 2017 Sep.
[Is] ISSN:1872-7905
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:We developed a homogeneous bridging anti-drug antibody (ADA) assay on an electro chemiluminescent immunoassay (ECLIA) platform to support the immunogenicity evaluation of a dimeric domain antibody (dAb) therapeutic in clinical studies. During method development we evaluated the impact of different types of acid at various pH levels on polyclonal and monoclonal ADA controls of differing affinities and on/off rates. The data shows for the first time that acids of different pH can have a differential effect on ADA of various affinities and this in turn impacts assay sensitivity and drug tolerance as defined by these surrogate controls. Acid treatment led to a reduction in signal of intermediate and low affinity ADA, but not high affinity or polyclonal ADA. We also found that acid pretreatment is a requisite for dissociation of drug bound high affinity ADA, but not for low affinity ADA-drug complexes. Although we were unable to identify an acid that would allow a 100% retrieval of ADA signal post-treatment, use of glycine pH3.0 enabled the detection of low, intermediate and high affinity antibodies (Abs) to various extents. Following optimization, the ADA assay method was validated for clinical sample analysis. Consistencies within various parameters of the clinical data such as dose dependent increases in ADA rates and titers were observed, indicating a reliable ADA method. Pre- and post-treatment ADA negative or positive clinical samples without detectable drug were reanalyzed in the absence of acid treatment or presence of added exogenous drug respectively to further assess the effectiveness of the final acid treatment procedure. The overall ADA results indicate that assay conditions developed and validated based on surrogate controls sufficed to provide a reliable clinical data set. The effect of low pH acid treatment on possible pre-existing ADA or soluble multimeric target in normal human serum was also evaluated, and preliminary data indicate that acid type and pH also affect drug-specific signal differentially in individual samples. The results presented here represent the most extensive analyses to date on acid treatment of a wide range of ADA affinities to explore sensitivity and drug tolerance issues. They have led to a refinement of our current best practices for ADA method development and provide a depth of data to interrogate low pH mediated immune complex dissociation.
[Mh] Termos MeSH primário: Ácidos/química
Anticorpos Anti-Idiotípicos/imunologia
Anticorpos Monoclonais/imunologia
Antineoplásicos/imunologia
Técnicas Eletroquímicas
Imunoensaio/métodos
[Mh] Termos MeSH secundário: Animais
Anticorpos Anti-Idiotípicos/sangue
Anticorpos Anti-Idiotípicos/química
Anticorpos Monoclonais/efeitos adversos
Anticorpos Monoclonais/sangue
Anticorpos Monoclonais/química
Afinidade de Anticorpos
Especificidade de Anticorpos
Antineoplásicos/efeitos adversos
Antineoplásicos/sangue
Antineoplásicos/química
Sítios de Ligação de Anticorpos
Estabilidade de Medicamentos
Glicina/química
Seres Humanos
Concentração de Íons de Hidrogênio
Camundongos Endogâmicos BALB C
Valor Preditivo dos Testes
Ligação Proteica
Desnaturação Proteica
Estabilidade Proteica
Reprodutibilidade dos Testes
[Pt] Tipo de publicação:JOURNAL ARTICLE; VALIDATION STUDIES
[Nm] Nome de substância:
0 (Acids); 0 (Antibodies, Anti-Idiotypic); 0 (Antibodies, Monoclonal); 0 (Antineoplastic Agents); TE7660XO1C (Glycine)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170918
[Lr] Data última revisão:
170918
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170620
[St] Status:MEDLINE


  10 / 12988 MEDLINE  
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[PMID]:28619012
[Au] Autor:Hosogai M; Nakatani Y; Mimura K; Kishi S; Akiyama H
[Ad] Endereço:Department of Ophthalmology, Gunma University Graduate School of Medicine, 3-39-22 Showa-machi, Maebashi, Gunma, 371-8511, Japan. mayu64jun@gmail.com.
[Ti] Título:Genetic analysis of varicella-zoster virus in the aqueous humor in uveitis with severe hyphema.
[So] Source:BMC Infect Dis;17(1):427, 2017 Jun 15.
[Is] ISSN:1471-2334
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Genetic variations have been identified in the genome of varicella-zoster virus (VZV) strains using vesicle fluid, varicella scabs and throat swab samples. We report a rare case of VZV-associated uveitis with severe hyphema, which was immediately diagnosed by polymerase chain reaction (PCR) using the aqueous humor, in which we were able to analyze the VZV genotype for the first time. CASE PRESENTATION: A 16-year-old Japanese boy was referred to our hospital with a 20-day history of unilateral anterior uveitis and 11-day history of hyphema. At presentation, details of the iris, the iridocorneal angle, and the fundus were not visible due to the severe hyphema. Serum anti-VZV IgG and anti-VZV IgM were elevated, and 1.61 × 10 copies/mL of VZV-DNA were detected by real-time PCR using the aqueous humor. As there were no eruptions on his face or body, we diagnosed zoster sine herpete and started intravenous administration of prednisolone and acyclovir. The hyphema completely disappeared 2 weeks after presentation, while sectorial iris atrophy and mild periphlebitis of the fundus became gradually apparent. Anterior inflammation and periphlebitis gradually improved and VZV-DNA in the aqueous humor was reduced to 1.02 × 10 copies/mL at 4 weeks after presentation. Examination by slit lamp microscope revealed no inflammation after 5 months, and VZV-DNA could no longer be detected in the aqueous humor. Serum anti-VZV IgG and anti-VZV IgM also showed a gradual decrease along with improvement in ocular inflammation. The genetic analysis of multiple open reading frames and the R5 variable repeat region in the VZV genes, using DNA extracted from the aqueous humor at presentation, showed that the isolate was a wild-type clade 2 VZV strain (prevalent in Japan and surrounding countries) with R5A allele and one SNP unique to clade 1 (both are major types in Europe and North America). CONCLUSIONS: VZV-associated uveitis may develop hyphema that obscures ocular inflammation, thus PCR analysis using the aqueous humor is the key investigation necessary for the diagnosis. The measurement of VZV-DNA copies by real-time PCR would be useful for evaluation of therapeutic effects. We could amplify and analyze VZV genotype using the aqueous humor including a very large number of VZV-DNA copies (1.61 × 10 copies/mL).
[Mh] Termos MeSH primário: Humor Aquoso/virologia
Herpes Zoster Oftálmico/complicações
Herpesvirus Humano 3/genética
Hifema/virologia
Uveíte Anterior/virologia
[Mh] Termos MeSH secundário: Aciclovir/uso terapêutico
Adolescente
Anticorpos Anti-Idiotípicos/sangue
DNA Viral/análise
Europa (Continente)
Genótipo
Herpes Zoster Oftálmico/diagnóstico
Herpes Zoster Oftálmico/tratamento farmacológico
Herpesvirus Humano 3/isolamento & purificação
Herpesvirus Humano 3/patogenicidade
Seres Humanos
Japão
Masculino
Reação em Cadeia da Polimerase
Uveíte Anterior/tratamento farmacológico
[Pt] Tipo de publicação:CASE REPORTS; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antibodies, Anti-Idiotypic); 0 (DNA, Viral); 0 (anti-IgM); X4HES1O11F (Acyclovir)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171002
[Lr] Data última revisão:
171002
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170617
[St] Status:MEDLINE
[do] DOI:10.1186/s12879-017-2518-2



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