Application of area scaling analysis to identify natural killer cell and monocyte involvement in the GranToxiLux antibody dependent cell-mediated cytotoxicity assay.
Abstract
Several different assay methodologies have been described for the evaluation of HIV
or SIV-specific antibody-dependent cell-mediated cytotoxicity (ADCC). Commonly used
assays measure ADCC by evaluating effector cell functions, or by detecting elimination
of target cells. Signaling through Fc receptors, cellular activation, cytotoxic granule
exocytosis, or accumulation of cytolytic and immune signaling factors have been used
to evaluate ADCC at the level of the effector cells. Alternatively, assays that measure
killing or loss of target cells provide a direct assessment of the specific killing
activity of antibodies capable of ADCC. Thus, each of these two distinct types of
assays provides information on only one of the critical components of an ADCC event;
either the effector cells involved, or the resulting effect on the target cell. We
have developed a simple modification of our previously described high-throughput ADCC
GranToxiLux (GTL) assay that uses area scaling analysis (ASA) to facilitate simultaneous
quantification of ADCC activity at the target cell level, and assessment of the contribution
of natural killer cells and monocytes to the total observed ADCC activity when whole
human peripheral blood mononuclear cells are used as a source of effector cells. The
modified analysis method requires no additional reagents and can, therefore, be easily
included in prospective studies. Moreover, ASA can also often be applied to pre-existing
ADCC-GTL datasets. Thus, incorporation of ASA to the ADCC-GTL assay provides an ancillary
assessment of the ability of natural and vaccine-induced antibodies to recruit natural
killer cells as well as monocytes against HIV or SIV; or to any other field of research
for which this assay is applied. © 2018 The Authors. Cytometry Part A published by
Wiley Periodicals, Inc. on behalf of ISAC.
Type
Journal articleSubject
GranToxiLux assayHIV
antibody-dependent cell-mediated cytotoxicity
monocytes
natural killer cells
peripheral blood mononuclear cells
Permalink
https://hdl.handle.net/10161/17800Published Version (Please cite this version)
10.1002/cyto.a.23348Publication Info
Pollara, Justin; Orlandi, Chiara; Beck, Charles; Edwards, R Whitney; Hu, Yi; Liu,
Shuying; ... Ferrari, Guido (2018). Application of area scaling analysis to identify natural killer cell and monocyte
involvement in the GranToxiLux antibody dependent cell-mediated cytotoxicity assay.
Cytometry. Part A : the journal of the International Society for Analytical Cytology, 93(4). pp. 436-447. 10.1002/cyto.a.23348. Retrieved from https://hdl.handle.net/10161/17800.This is constructed from limited available data and may be imprecise. To cite this
article, please review & use the official citation provided by the journal.
Collections
More Info
Show full item recordScholars@Duke
Thomas Norton Denny
Professor in Medicine
Thomas N. Denny, MSc, M.Phil, is the Chief Operating Officer of the Duke Human Vaccine
Institute (DHVI), Associate Dean for Duke Research and Discovery @RTP, and a Professor
of Medicine in the Department of Medicine at Duke University Medical Center. He is
also an Affiliate Member of the Duke Global Health Institute. Previously, he served
on the Health Sector Advisory Council of the Duke University Fuquay School of Business.
Prior to joining Duke, he was an Associate Professor of Pathology, Labo
Guido Ferrari
Professor in Surgery
The activities of the Ferrari Laboratory are based on both independent basic research
and immune monitoring studies. The research revolves around three main areas of interest:
class I-mediated cytotoxic CD8+ T cell responses, antibody-dependent cellular cytotoxicity
(ADCC), gene expression in NK and T cellular subsets upon infection with HIV-1. With
continuous funding over the last 11 years from the NIH and Bill & Melinda Gates Foundation
along with many other productive collaborations wi
Justin Joseph Pollara
Associate Professor in Surgery
Georgia Doris Tomaras
Professor in Surgery
Dr. Georgia Tomaras is a tenured Professor of Surgery, Professor of Immunology, Professor
of Molecular Genetics and Microbiology and is a Fellow of the American Academy of
Microbiology (AAM) and a Fellow of the American Association for the Advancement of
Science (AAAS). Dr. Tomaras is Co-Director of the Center for Human Systems Immunology
(CHSI) Duke University and Director of the Duke Center for AIDS Research (CFAR). Her
national and international leadership roles i
Alphabetical list of authors with Scholars@Duke profiles.

Articles written by Duke faculty are made available through the campus open access policy. For more information see: Duke Open Access Policy
Rights for Collection: Scholarly Articles
Works are deposited here by their authors, and represent their research and opinions, not that of Duke University. Some materials and descriptions may include offensive content. More info