RESUMO
Coronavirus disease 2019 (COVID-19) exhibits variable symptom severity ranging from asymptomatic to life-threatening, yet the relationship between severity and the humoral immune response is poorly understood. We examined antibody responses in 113 COVID-19 patients and found that severe cases resulting in intubation or death exhibited increased inflammatory markers, lymphopenia, pro-inflammatory cytokines, and high anti-receptor binding domain (RBD) antibody levels. Although anti-RBD immunoglobulin G (IgG) levels generally correlated with neutralization titer, quantitation of neutralization potency revealed that high potency was a predictor of survival. In addition to neutralization of wild-type SARS-CoV-2, patient sera were also able to neutralize the recently emerged SARS-CoV-2 mutant D614G, suggesting cross-protection from reinfection by either strain. However, SARS-CoV-2 sera generally lacked cross-neutralization to a highly homologous pre-emergent bat coronavirus, WIV1-CoV, which has not yet crossed the species barrier. These results highlight the importance of neutralizing humoral immunity on disease progression and the need to develop broadly protective interventions to prevent future coronavirus pandemics.
Assuntos
Anticorpos Neutralizantes/imunologia , Biomarcadores/análise , COVID-19/imunologia , COVID-19/fisiopatologia , Adulto , Anticorpos Neutralizantes/análise , Anticorpos Antivirais/análise , Anticorpos Antivirais/sangue , Biomarcadores/sangue , COVID-19/sangue , COVID-19/epidemiologia , Comorbidade , Coronavirus/classificação , Coronavirus/fisiologia , Reações Cruzadas , Citocinas/sangue , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imunoglobulina A/análise , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Imunoglobulina M/sangue , Imunoglobulina M/imunologia , Masculino , Massachusetts/epidemiologia , Pessoa de Meia-Idade , Domínios Proteicos , SARS-CoV-2/química , SARS-CoV-2/fisiologia , Índice de Gravidade de Doença , Glicoproteína da Espícula de Coronavírus/química , Análise de Sobrevida , Resultado do TratamentoAssuntos
Acidentes por Quedas , Disfunção Cognitiva , Hiperestesia , Limitação da Mobilidade , Idoso de 80 Anos ou mais , Feminino , Humanos , Disfunção Cognitiva/etiologia , Hiperestesia/etiologia , Tato , Pé , Joelho , MãosAssuntos
Confusão , Discinesias , Idoso , Feminino , Humanos , Confusão/etiologia , Discinesias/etiologiaRESUMO
The diagnosis of COVID-19 requires integration of clinical and laboratory data. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) diagnostic assays play a central role in diagnosis and have fixed technical performance metrics. Interpretation becomes challenging because the clinical sensitivity changes as the virus clears and the immune response emerges. Our goal was to examine the clinical sensitivity of two most common SARS-CoV-2 diagnostic test modalities, polymerase chain reaction (PCR) and serology, over the disease course to provide insight into their clinical interpretation in patients presenting to the hospital. We conducted a single-center, retrospective study. To derive clinical sensitivity of PCR, we identified 209 PCR-positive SARS-CoV-2 patients with multiple PCR test results (624 total PCR tests) and calculated daily sensitivity from date of symptom onset or first positive test. Clinical sensitivity of PCR decreased with days post symptom onset with >90% clinical sensitivity during the first 5 days after symptom onset, 70%-71% from Days 9 to 11, and 30% at Day 21. To calculate daily clinical sensitivity by serology, we utilized 157 PCR-positive patients with a total of 197 specimens tested by enzyme-linked immunosorbent assay for IgM, IgG, and IgA anti-SARS-CoV-2 antibodies. In contrast to PCR, serological sensitivity increased with days post symptom onset with >50% of patients seropositive by at least one antibody isotype after Day 7, >80% after Day 12, and 100% by Day 21. Taken together, PCR and serology are complimentary modalities that require time-dependent interpretation. Superimposition of sensitivities over time indicate that serology can function as a reliable diagnostic aid indicating recent or prior infection.
Assuntos
Teste de Ácido Nucleico para COVID-19 , Teste Sorológico para COVID-19 , COVID-19/diagnóstico , SARS-CoV-2 , Anticorpos Antivirais/sangue , COVID-19/sangue , Feminino , Hospitais , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Sensibilidade e EspecificidadeRESUMO
SARS-CoV-2 antibody testing allows quantitative determination of disease prevalence, which is especially important in high-risk communities. We performed anonymized convenience sampling of 200 currently asymptomatic residents of Chelsea, the epicenter of COVID-19 illness in Massachusetts, by BioMedomics SARS-CoV-2 combined IgM-IgG point-of-care lateral flow immunoassay. The seroprevalence was 31.5% (17.5% IgM+IgG+, 9.0% IgM+IgG-, and 5.0% IgM-IgG+). Of the 200 participants, 50.5% reported no symptoms in the preceding 4 weeks, of which 24.8% (25/101) were seropositive, and 60% of these were IgM+IgG-. These data are the highest seroprevalence rates observed to date and highlight the significant burden of asymptomatic infection.
Assuntos
Anticorpos Antivirais/sangue , COVID-19/diagnóstico , Sistemas Automatizados de Assistência Junto ao Leito , Adulto , Especificidade de Anticorpos , COVID-19/epidemiologia , COVID-19/virologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imunoensaio , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Masculino , Massachusetts/epidemiologia , Pessoa de Meia-Idade , Análise Multivariada , Análise de Regressão , Estudos SoroepidemiológicosAssuntos
Proteína Inibidora do Complemento C1/análise , Angioedema Hereditário Tipos I e II/diagnóstico , Anticorpos Monoclonais Humanizados/uso terapêutico , Proteínas do Sistema Complemento/análise , Diagnóstico Diferencial , Edema/etiologia , Feminino , Deleção de Genes , Angioedema Hereditário Tipos I e II/tratamento farmacológico , Angioedema Hereditário Tipos I e II/genética , Humanos , Pessoa de Meia-Idade , Radiografia Abdominal , Tomografia Computadorizada por Raios X , Urticária/etiologiaRESUMO
The complement pathway is a cascade of proteases that is involved in immune surveillance and innate immunity, as well as adaptive immunity. Dysfunction of the complement cascade may be mediated by aberrations in the pathways of activation, complement regulatory proteins, or complement deficiencies, and has been linked to a number of hematologic disorders, including paroxysmal noctural hemoglobinuria (PNH), hereditary angioedema (HAE), and atypical hemolytic-uremic syndrome (aHUS). Here, current laboratory tests for disorders of the complement pathway are reviewed, and their utility and limitations in hematologic disorders and systemic diseases are discussed. Current therapeutic advances targeting the complement pathway in treatment of complement-mediated hematologic disorders are also reviewed.
Assuntos
Serviços de Laboratório Clínico/estatística & dados numéricos , Ativação do Complemento/imunologia , Proteínas do Sistema Complemento/imunologia , HumanosAssuntos
Anafilaxia/etiologia , Equinococose Hepática/diagnóstico , Echinococcus granulosus/isolamento & purificação , Fígado/patologia , Fígado/parasitologia , Dor Abdominal/etiologia , Adulto , Anafilaxia/diagnóstico , Animais , Diagnóstico Diferencial , Equinococose Hepática/complicações , Equinococose Hepática/imunologia , Humanos , Hipotensão/etiologia , Masculino , Síncope/etiologia , Tomografia Computadorizada por Raios XRESUMO
Genetic characterization of myeloma at diagnosis by interphase fluorescence in situ hybridization and next-generation sequencing (NGS) can assist with risk stratification and treatment planning. Measurable residual disease (MRD) status after treatment, as evaluated by next-generation flow cytometry or NGS on bone marrow aspirate material, is one of the most important predictors of prognosis. Less-invasive tools for MRD assessment such as liquid biopsy approaches have also recently emerged as potential alternatives.
Assuntos
Mieloma Múltiplo , Plasmocitoma , Humanos , Mieloma Múltiplo/diagnóstico , Mieloma Múltiplo/genética , Patologia Molecular , Hibridização in Situ Fluorescente , Prognóstico , Neoplasia Residual/diagnóstico , Neoplasia Residual/genética , Neoplasia Residual/patologia , Sequenciamento de Nucleotídeos em Larga Escala , Citometria de FluxoRESUMO
OBJECTIVES: There is considerable variation in ordering practices for the initial laboratory evaluation of monoclonal gammopathies (MGs) despite clear society guidelines to include serum free light chain (sFLC) testing. We assessed the ability of a clinical decision support (CDS) alert to improve guideline compliance and analyzed its clinical impact. METHODS: We designed and deployed a targeted CDS alert to educate and prompt providers to order an sFLC assay when ordering serum protein electrophoresis (SPEP) testing. RESULTS: The alert was highly effective at increasing the co-ordering of SPEP and sFLC testing. Preimplementation, 62.8% of all SPEP evaluations included sFLC testing, while nearly 90% of evaluations included an sFLC assay postimplementation. In patients with no prior sFLC testing, analysis of sFLC orders prompted by the alert led to the determination that 28.9% (800/2,769) of these patients had an abnormal κ/λ ratio. In 452 of these patients, the sFLC assay provided the only laboratory evidence of a monoclonal protein. Moreover, within this population, there were numerous instances of new diagnoses of multiple myeloma and other MGs. CONCLUSIONS: The CDS alert increased compliance with society guidelines and improved the diagnostic evaluation of patients with suspected MGs.
Assuntos
Sistemas de Apoio a Decisões Clínicas , Mieloma Múltiplo , Paraproteinemias , Humanos , Paraproteinemias/diagnóstico , Cadeias Leves de ImunoglobulinaAssuntos
Hipertensão Pulmonar/etiologia , Linfonodos/patologia , Doença Mista do Tecido Conjuntivo/patologia , Adulto , Anticorpos Antinucleares/sangue , Diagnóstico Diferencial , Dispneia/etiologia , Feminino , Humanos , Hipertensão Pulmonar/diagnóstico , Hipertrofia Ventricular Esquerda/diagnóstico por imagem , Doença Mista do Tecido Conjuntivo/complicações , Derrame Pericárdico/diagnóstico por imagem , Radiografia Torácica , Doença de Raynaud/complicações , Ribonucleoproteína Nuclear Pequena U1/imunologia , UltrassonografiaAssuntos
Hepatite B/diagnóstico , Doença do Soro/diagnóstico , Doença Aguda , Adulto , Artrite/etiologia , Diagnóstico Diferencial , Exantema/etiologia , Febre/etiologia , Hepatite B/complicações , Dependência de Heroína/complicações , Humanos , Masculino , Dor Musculoesquelética/etiologia , Doença do Soro/etiologia , Doença do Soro/imunologia , Abuso de Substâncias por Via Intravenosa/complicaçõesAssuntos
Alveolite Alérgica Extrínseca/patologia , Aspergillus flavus , Aspergillus fumigatus , Pulmão/patologia , Alveolite Alérgica Extrínseca/complicações , Alveolite Alérgica Extrínseca/diagnóstico por imagem , Dor no Peito/etiologia , Criança , Tosse/etiologia , Diagnóstico Diferencial , Dispneia/etiologia , Febre/etiologia , Humanos , Pulmão/diagnóstico por imagem , Pneumopatias/diagnóstico , Doenças Linfáticas/diagnóstico por imagem , Masculino , Radiografia , UltrassonografiaRESUMO
In a variety of hematologic malignancies, immunoglobulin light chains (LC) are overproduced clonally and circulate without being linked by disulphide bonds to the immunoglobulin heavy chain. The recent development of a robust assay known as κ and λ "free" LC (FLC) to quantify the levels of these unbound LC in the serum, and thereby determine their ratio, has led to an explosion of studies that demonstrate its utility in a wide range of hematologic disorders. This article summarizes laboratory testing for serum FLC, with a particular focus on clinical applications for the test.
Assuntos
Cadeias kappa de Imunoglobulina/sangue , Cadeias lambda de Imunoglobulina/sangue , Nefelometria e Turbidimetria/métodos , Algoritmos , Amiloidose/sangue , Amiloidose/diagnóstico , Anticorpos Monoclonais/sangue , Proteína de Bence Jones/urina , Eletroforese das Proteínas Sanguíneas , Densitometria , Meia-Vida , Neoplasias Hematológicas/sangue , Humanos , Cadeias kappa de Imunoglobulina/análise , Cadeias lambda de Imunoglobulina/análise , Proteínas do Mieloma/análise , Paraproteinemias/sangue , Paraproteinemias/diagnóstico , Paraproteinemias/urina , Ligação Proteica , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , SoroRESUMO
Critical to managing the spread of COVID-19 is the ability to diagnose infection and define the acquired immune response across the population. While genomic tests for the novel Several Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) detect the presence of viral RNA for a limited time frame, when the virus is shed in the upper respiratory tract, tests able to define exposure and infection beyond this short window of detectable viral replication are urgently needed. Following infection, antibodies are generated within days, providing a durable read-out and archive of exposure and infection. Several antibody tests have emerged to diagnose SARS-CoV-2. Here we report on a qualified quantitative ELISA assay that displays all the necessary characteristics for high-throughput sample analysis. Collectively, this test offers a quantitative opportunity to define both exposure and levels of immunity to SARS-CoV-2.
Assuntos
Betacoronavirus/isolamento & purificação , Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/diagnóstico , Ensaio de Imunoadsorção Enzimática , Pneumonia Viral/diagnóstico , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Anticorpos Antivirais/isolamento & purificação , Betacoronavirus/imunologia , COVID-19 , Teste para COVID-19 , Infecções por Coronavirus/sangue , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/virologia , Estudos de Viabilidade , Ensaios de Triagem em Larga Escala , Humanos , Pandemias , Pneumonia Viral/sangue , Pneumonia Viral/imunologia , Pneumonia Viral/virologia , Reprodutibilidade dos Testes , SARS-CoV-2 , Sensibilidade e Especificidade , Fatores de TempoRESUMO
COVID-19 exhibits variable symptom severity ranging from asymptomatic to life-threatening, yet the relationship between severity and the humoral immune response is poorly understood. We examined antibody responses in 113 COVID-19 patients and found that severe cases resulting in intubation or death exhibited increased inflammatory markers, lymphopenia, and high anti-RBD antibody levels. While anti-RBD IgG levels generally correlated with neutralization titer, quantitation of neutralization potency revealed that high potency was a predictor of survival. In addition to neutralization of wild-type SARS-CoV-2, patient sera were also able to neutralize the recently emerged SARS-CoV-2 mutant D614G, suggesting protection from reinfection by this strain. However, SARS-CoV-2 sera was unable to cross-neutralize a highly-homologous pre-emergent bat coronavirus, WIV1-CoV, that has not yet crossed the species barrier. These results highlight the importance of neutralizing humoral immunity on disease progression and the need to develop broadly protective interventions to prevent future coronavirus pandemics.
Assuntos
Medula Óssea/patologia , Mastocitose Sistêmica/patologia , Adulto , Anafilaxia/diagnóstico , Diagnóstico Diferencial , Ecocardiografia , Eletrocardiografia , Neoplasias das Glândulas Endócrinas/diagnóstico , Rubor/etiologia , Humanos , Hipotensão/etiologia , Masculino , Mastocitose Sistêmica/complicações , Mastocitose Sistêmica/diagnóstico , Radiografia , Costelas/diagnóstico por imagem , Costelas/patologia , Vértebras Torácicas/diagnóstico por imagem , Vértebras Torácicas/patologiaRESUMO
Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by pathogenic autoantibodies against nucleoproteins and DNA. Here we show that DNA-containing immune complexes (ICs) within lupus serum (SLE-ICs), but not protein-containing ICs from other autoimmune rheumatic diseases, stimulates plasmacytoid DCs (PDCs) to produce cytokines and chemokines via a cooperative interaction between Toll-like receptor 9 (TLR9) and FcgammaRIIa (CD32). SLE-ICs transiently colocalized to a subcellular compartment containing CD32 and TLR9, and CD32+, but not CD32-, PDCs internalized and responded to SLE-ICs. Our findings demonstrate a novel functional interaction between Fc receptors and TLRs, defining a pathway in which CD32 delivers SLE-ICs to intracellular lysosomes containing TLR9, inducing a signaling cascade leading to PDC activation. These data demonstrate that endogenous DNA-containing autoantibody complexes found in the serum of patients with SLE activate the innate immune system and suggest a novel mechanism whereby these ICs contribute to the pathogenesis of this autoimmune disease.