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A versatile high-throughput assay to characterize antibody-mediated neutrophil phagocytosis.
Karsten, Christina B; Mehta, Nickita; Shin, Sally A; Diefenbach, Thomas J; Slein, Matthew D; Karpinski, Wiktor; Irvine, Edward B; Broge, Thomas; Suscovich, Todd J; Alter, Galit.
Afiliação
  • Karsten CB; Ragon Institute of MGH, MIT and Harvard, 400 Technology Square, Cambridge, MA 02139, USA.
  • Mehta N; Ragon Institute of MGH, MIT and Harvard, 400 Technology Square, Cambridge, MA 02139, USA.
  • Shin SA; Ragon Institute of MGH, MIT and Harvard, 400 Technology Square, Cambridge, MA 02139, USA.
  • Diefenbach TJ; Ragon Institute of MGH, MIT and Harvard, 400 Technology Square, Cambridge, MA 02139, USA.
  • Slein MD; Ragon Institute of MGH, MIT and Harvard, 400 Technology Square, Cambridge, MA 02139, USA.
  • Karpinski W; Ragon Institute of MGH, MIT and Harvard, 400 Technology Square, Cambridge, MA 02139, USA.
  • Irvine EB; Ragon Institute of MGH, MIT and Harvard, 400 Technology Square, Cambridge, MA 02139, USA; Harvard T.H. Chan School of Public Health, 677 Huntington Ave, Boston, MA 02115, USA.
  • Broge T; Ragon Institute of MGH, MIT and Harvard, 400 Technology Square, Cambridge, MA 02139, USA.
  • Suscovich TJ; Ragon Institute of MGH, MIT and Harvard, 400 Technology Square, Cambridge, MA 02139, USA.
  • Alter G; Ragon Institute of MGH, MIT and Harvard, 400 Technology Square, Cambridge, MA 02139, USA. Electronic address: galter@mgh.harvard.edu.
J Immunol Methods ; 471: 46-56, 2019 08.
Article em En | MEDLINE | ID: mdl-31132351
ABSTRACT
Neutrophils, the most abundant white blood cell, play a critical role in anti-pathogen immunity via phagocytic clearance, secretion of enzymes and immunomodulators, and the release of extracellular traps. Neutrophils non-specifically sense infection through an array of innate immune receptors and inflammatory sensors, but are also able to respond in a pathogen/antigen-specific manner when leveraged by antibodies via Fc-receptors. Among neutrophil functions, antibody-dependent neutrophil phagocytosis (ADNP) results in antibody-mediated opsonization, enabling neutrophils to sense and respond to infection in a pathogen-appropriate manner. Here, we describe a high-throughput flow cytometric approach to effectively visualize and quantify ADNP and its downstream consequences. The assay is easily adaptable, supporting both the use of purified neutrophils or white blood cells, the use of purified Ig or serum, and the broad utility of any target antigen. Thus, this ADNP assay represents a high-throughput platform for the in-depth characterization of neutrophil function.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Fagocitose / Ensaios de Triagem em Larga Escala / Anticorpos / Complexo Antígeno-Anticorpo / Neutrófilos Limite: Humans Idioma: En Revista: J Immunol Methods Ano de publicação: 2019 Tipo de documento: Article País de afiliação: Estados Unidos
Texto completo
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Fagocitose / Ensaios de Triagem em Larga Escala / Anticorpos / Complexo Antígeno-Anticorpo / Neutrófilos Limite: Humans Idioma: En Revista: J Immunol Methods Ano de publicação: 2019 Tipo de documento: Article País de afiliação: Estados Unidos