A robust, high-throughput assay to determine the phagocytic activity of clinical antibody samples
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Author(s)
Ackerman, Margaret E.
Moldt, Brian
Wyatt, Richard T.
Dugast, Anne-Sophie
McAndrew, Elizabeth G.
Tsoukas, Stephen
Jost, Stephanie
Berger, Christoph T.
Sciaranghella, Gaia
Liu, Qingquan
Date Issued
December 2010
Journal
Journal of Immunological Methods
Publisher
Elsevier
Citation
Ackerman, Margaret E., Brian Moldt, Richard T. Wyatt, Anne-Sophie Dugast, Elizabeth McAndrew, Stephen Tsoukas, Stephanie Jost, et al. “A Robust, High-Throughput Assay to Determine the Phagocytic Activity of Clinical Antibody Samples.” Journal of Immunological Methods 366, no. 1–2 (March 2011): 8–19.
Version
Author's final manuscript
Abstract
Phagocytosis can be induced via the engagement of Fcγ receptors by antibody-opsonized material. Furthermore, the efficiency of antibody-induced effector functions has been shown to be dramatically modulated by changes in antibody glycosylation. Because infection can modulate antibody glycans, which in turn modulate antibody functions, assays capable of determining the induction of effector functions rather than neutralization or titer provide a valuable opportunity to more fully characterize the quality of the adaptive immune response. Here we describe a robust and high-throughput flow cytometric assay to define the phagocytic activity of antigen-specific antibodies from clinical samples. This assay employs a monocytic cell line that expresses numerous Fc receptors: including inhibitory and activating, and high and low affinity receptors—allowing complex phenotypes to be studied. We demonstrate the adaptability of this high-throughput, flow-based assay to measure antigen-specific antibody-mediated phagocytosis against an array of viruses, including influenza, HIV, and dengue. The phagocytosis assay format further allows for simultaneous analysis of cytokine release, as well as determination of the role of specific Fcγ-receptor subtypes, making it a highly useful system for parsing differences in the ability of clinical and vaccine induced antibody samples to recruit this critical effector function.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Materials Science and Engineering
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DOI of Published Version
https://doi.org/10.1016/j.jim.2010.12.016
