RESUMO
Potent neutralizing SARS-CoV-2 antibodies often target the spike protein receptor-binding site (RBS), but the variability of RBS epitopes hampers broad neutralization of multiple sarbecoviruses and drifted viruses. Here, using humanized mice, we identified an RBS antibody with a germline VH gene that potently neutralized SARS-related coronaviruses, including SARS-CoV and SARS-CoV-2 variants. X-ray crystallography revealed coordinated recognition by the heavy chain of non-RBS conserved sites and the light chain of RBS with a binding angle mimicking the angiotensin-converting enzyme 2 (ACE2) receptor. The minimum footprints in the hypervariable region of RBS contributed to the breadth of neutralization, which was enhanced by immunoglobulin G3 (IgG3) class switching. The coordinated binding resulted in broad neutralization of SARS-CoV and emerging SARS-CoV-2 variants of concern. Low-dose therapeutic antibody treatment in hamsters reduced the virus titers and morbidity during SARS-CoV-2 challenge. The structural basis for broad neutralizing activity may inform the design of a broad spectrum of therapeutics and vaccines.
Assuntos
Anticorpos Amplamente Neutralizantes/imunologia , Reações Cruzadas/imunologia , SARS-CoV-2/imunologia , Animais , Betacoronavirus/imunologia , Sítios de Ligação de Anticorpos , Anticorpos Amplamente Neutralizantes/química , Anticorpos Amplamente Neutralizantes/uso terapêutico , COVID-19/prevenção & controle , COVID-19/terapia , COVID-19/virologia , Cricetinae , Humanos , Switching de Imunoglobulina , Fragmentos Fab das Imunoglobulinas/química , Fragmentos Fab das Imunoglobulinas/metabolismo , Imunoglobulina G/química , Imunoglobulina G/imunologia , Camundongos , Domínios Proteicos , Glicoproteína da Espícula de Coronavírus/química , Glicoproteína da Espícula de Coronavírus/imunologia , Glicoproteína da Espícula de Coronavírus/metabolismoAssuntos
Adjuvantes Imunológicos/administração & dosagem , Analgésicos Opioides/imunologia , Comportamento Aditivo/prevenção & controle , Imunoconjugados/administração & dosagem , Imunoglobulina G/biossíntese , Transtornos Relacionados ao Uso de Opioides/prevenção & controle , Analgésicos Opioides/efeitos adversos , Comportamento Aditivo/imunologia , Sítios de Ligação de Anticorpos , Encéfalo/efeitos dos fármacos , Encéfalo/imunologia , Toxoide Diftérico/química , Toxoide Diftérico/imunologia , Haptenos/química , Haptenos/imunologia , Humanos , Imunoconjugados/química , Transtornos Relacionados ao Uso de Opioides/imunologia , Ovalbumina/química , Ovalbumina/imunologia , Ligação Proteica , Toxoide Tetânico/química , Toxoide Tetânico/imunologia , Pesquisa Translacional Biomédica/tendências , Vacinas Conjugadas/administração & dosagem , Vacinas Conjugadas/químicaRESUMO
The noncollagenous (NC1) domains of alpha3alpha4alpha5(IV) collagen in the glomerular basement membrane (GBM) are targets of Goodpasture autoantibodies or Alport posttransplant nephritis alloantibodies mediating rapidly progressive glomerulonephritis. Because the autoepitopes but not the alloepitopes become cryptic upon assembly of alpha3alpha4alpha5NC1 hexamers, we investigated how the accessibility of B cell epitopes in vivo influences the development of glomerulonephritis in mice passively immunized with human anti-GBM Abs. Alport alloantibodies, which bound to native murine alpha3alpha4alpha5NC1 hexamers in vitro, deposited linearly along the mouse GBM in vivo, eliciting crescentic glomerulonephritis in Fcgr2b(-/-) mice susceptible to Ab-mediated inflammation. Goodpasture autoantibodies, which bound to murine alpha3NC1 monomer and dimer subunits but not to native alpha3alpha4alpha5NC1 hexamers in vitro, neither bound to the mouse GBM in vivo nor induced experimental glomerulonephritis. This was due to quinary NC1 crosslinks, recently identified as sulfilimine bonds, which comprehensively locked the cryptic Goodpasture autoepitopes in the mouse GBM. In contrast, non-crosslinked alpha3NC1 subunits were identified as a native target of Goodpasture autoantibodies in the GBM of squirrel monkeys, a species susceptible to Goodpasture autoantibody-mediated nephritis. Thus, crypticity of B cell autoepitopes in tissues uncouples potentially pathogenic autoantibodies from autoimmune disease. Crosslinking of alpha3alpha4alpha5NC1 hexamers represents a novel mechanism averting autoantibody binding and subsequent tissue injury by posttranslational modifications of an autoantigen.
Assuntos
Doença Antimembrana Basal Glomerular/imunologia , Autoanticorpos/fisiologia , Autoantígenos/imunologia , Colágeno Tipo IV/imunologia , Epitopos/imunologia , Glomerulonefrite/imunologia , Isoanticorpos/fisiologia , Nefrite Hereditária/imunologia , Animais , Autoanticorpos/metabolismo , Autoantígenos/metabolismo , Sítios de Ligação de Anticorpos , Colágeno Tipo IV/metabolismo , Reações Cruzadas/imunologia , Epitopos/metabolismo , Glomerulonefrite/metabolismo , Glomerulonefrite/prevenção & controle , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Estrutura Terciária de ProteínaRESUMO
Interferon beta (IFN-beta) is generally considered an effective treatment for multiple sclerosis (MS). Importance of binding antibodies (BAb), which are created during the treatment of MS by the use of IFN-beta, hasn't been completely explained, however it is generally reckoned that they might be one of the factors diminishing treatment efficacy. The aim of the study was the appreciation of BAb occurrence during the treatment of MS by the use of different types of interferon beta and their impact on clinical efficacy. The study included 47 patients with relapsing-remitting MS. Within 24 months 37 patients were given two different preparations of IFN-beta 1-a and 10 patients were given IFN-beta 1-b. Every 6 months clinical parameters and BAb level in serum by EIA method were estimated. All preparations of IFN-beta induced appearance of BAb, but frequency of developing BAb to IFN-beta varied according to the IFN beta given. The high levels of BAb appeared significant frequently in patients treated with IFN-beta 1-b than in patients treated with both preparations of IFN-beta 1-a. After 2 years of treatment greater disability, measured by EDSS scale was encountered in patients with high levels of BAb but differences weren't statistically significant. As well, it wasn't stated significant correlation between exacerbation numbers during the treatment.
Assuntos
Sítios de Ligação de Anticorpos/efeitos dos fármacos , Interferon beta/administração & dosagem , Esclerose Múltipla Recidivante-Remitente/tratamento farmacológico , Esclerose Múltipla Recidivante-Remitente/imunologia , Adjuvantes Imunológicos/administração & dosagem , Adulto , Feminino , Humanos , Interferon beta-1a , Interferon beta-1b , Masculino , Resultado do TratamentoAssuntos
Rubéola (Sarampo Alemão)/prevenção & controle , Fatores Etários , Formação de Anticorpos , Sítios de Ligação de Anticorpos , Proteínas do Sistema Complemento , Feminino , Doenças Fetais/etiologia , Humanos , Itália , Gravidez , Complicações Infecciosas na Gravidez/prevenção & controle , Rubéola (Sarampo Alemão)/epidemiologia , Rubéola (Sarampo Alemão)/imunologia , Vacina contra RubéolaRESUMO
To assess whether an abnormality in transferrin receptor expression or regulation could represent an underlying metabolic defect in the reticuloendothelial (RE) system in hemochromatosis, monocytes were analyzed for the expression of the transferrin receptor using a monoclonal antibody (Act II) to the transferrin receptor (CD71) and flow cytometric analysis. Hemochromatosis patients (n = 14), and normal volunteers with no clinical evidence of iron overload (n = 14) were studied. A significant inverse relationship was observed for the relationship between the expression of transferrin receptor on monocytes and log(hepatic iron concentration) in hemochromatosis patients (r = -0.59, P less than .02) and also for the relationship between the expression of transferrin receptor and log(serum ferritin) in normal volunteers (r = -0.90, P less than .001). There was no significant difference in the mean expression of monocyte transferrin receptor between hemochromatosis patients and normal volunteers. However, the expression of the transferrin receptor in hemochromatosis patients was disproportionately higher than would be predicted from the relationship between serum ferritin and transferrin receptor expression in normal volunteers. The inverse relationship of the monocyte transferrin receptor relative to body iron stores in hemochromatosis is consistent with observations in other tissues, and suggests that non-transferrin iron metabolism, including ferritin, requires further investigation in the RE cell in hemochromatosis.