A versatile high-throughput assay to characterize antibody-mediated neutrophil phagocytosis

被引：116

作者：

Karsten, Christina B. ^{[1
]}

Mehta, Nickita ^{[1
]}

Shin, Sally A. ^{[1
]}

Diefenbach, Thomas J. ^{[1
]}

Slein, Matthew D. ^{[1
]}

Karpinski, Wiktor ^{[1
]}

Irvine, Edward B. ^{[1
,2
]}

Broge, Thomas ^{[1
]}

Suscovich, Todd J. ^{[1
]}

Alter, Galit ^{[1
]}

机构：

[1] Ragon Inst MGH MIT & Harvard, 400 Technol Sq, Cambridge, MA 02139 USA

[2] Harvard TH Chan Sch Publ Hlth, 677 Huntington Ave, Boston, MA 02115 USA

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 2019年 / 471卷

关键词：

Non-neutralizing antibody; Antibody-dependent neutrophil phagocytosis; Systems serology; High-throughput assay; Immune correlates of protection; EXTRACELLULAR TRAPS; PROTECTIVE EFFICACY; IN-VITRO; ACTIVATION; EXPRESSION; AFFINITY; VACCINE; CELLS; QUANTIFICATION; MODULATION;

D O I：

10.1016/j.jim.2019.05.006

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：46 / 56

页数：11

共 54 条

[21]

DRESCH C, 1971, J NUCL MED, V12, P774

[22] Measurement of phagosomal pH of normal and CGD-Like human neutrophils by dual fluorescence flow cytometry [J].

Dri, P ;

Presani, G ;

Perticarari, S ;

Albèri, L ;

Prodan, M ;

Decleva, E .

CYTOMETRY, 2002, 48 (03) :159-166

[23]

Engstad CS, 1997, THROMB HAEMOSTASIS, V77, P690

[24] Isolation and activation of human neutrophils in vitro.: The importance of the anticoagulant used during blood collection [J].

Freltas, Marisa ;

Porto, Graca ;

Lima, Jose L. F. C. ;

Femandes, Eduarda .

CLINICAL BIOCHEMISTRY, 2008, 41 (7-8) :570-575

[25] Neutrophils in viral infections: Current concepts and caveats [J].

Galani, Ioanna E. ;

Andreakos, Evangelos .

JOURNAL OF LEUKOCYTE BIOLOGY, 2015, 98 (04) :557-564

[26] Heterogeneity Among Neutrophils [J].

Garley, Marzena ;

Jablonska, Ewa .

ARCHIVUM IMMUNOLOGIAE ET THERAPIAE EXPERIMENTALIS, 2018, 66 (01) :21-30

[27] Flow cytometric assay for direct quantification of neutrophil extracellular traps in blood samples [J].

Gavillet, Mathilde ;

Martinod, Kimberly ;

Renella, Raffaele ;

Harris, Chad ;

Shapiro, Nate I. ;

Wagner, Denisa D. ;

Williams, David A. .

AMERICAN JOURNAL OF HEMATOLOGY, 2015, 90 (12) :1155-1158

[28] How neutrophils kill fungi [J].

Gazendam, Roel P. ;

van de Geer, Annemarie ;

Roos, Dirk ;

van den Berg, Timo K. ;

Kuijpers, Taco W. .

IMMUNOLOGICAL REVIEWS, 2016, 273 (01) :299-311

[29] A flow cytometry-based assay to determine the phagocytic activity of both clinical and nonclinical antibody samples against Chlamydia trachomatis [J].

Grasse, Marco ;

Rosenkrands, Ida ;

Olsen, Anja ;

Follmann, Frank ;

Dietrich, Jes .

CYTOMETRY PART A, 2018, 93A (05) :525-532

[30] Neutrophil Functional Heterogeneity: Identification of Competitive Phagocytosis [J].

Hellebrekers, Pien ;

Hietbrink, Falco ;

Vrisekoop, Nienke ;

Leenen, Luke P. H. ;

Koenderman, Leo .

FRONTIERS IN IMMUNOLOGY, 2017, 8

← 1 2 3 4 5 6 →