期刊
JOURNAL OF IMMUNOLOGICAL METHODS
卷 471, 期 -, 页码 46-56出版社
ELSEVIER
DOI: 10.1016/j.jim.2019.05.006
关键词
Non-neutralizing antibody; Antibody-dependent neutrophil phagocytosis; Systems serology; High-throughput assay; Immune correlates of protection
资金
- NIH [AI080289 (5R37AI080289-09), AI102660-01 (5R01AI102660-04), AI129797-01 (1R01A1129797-01)]
- Bill and Melinda Gates foundation [OPP1032817, OPP1114729, OPP1146996]
- CFAR Scholars Program [P30 AI060354-02]
- NIH AIDS Reagent Program Division of AIDS, NIAID, NIH [3957]
- T32 grant
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 nonspecifically 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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