Application of area scaling analysis to identify natural killer cell and monocyte involvement in the GranToxiLux antibody dependent cell-mediated cytotoxicity assay
JournalCytometry Part A
PublisherJohn Wiley and Sons
MetadataShow full item record
AbstractSeveral 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. Copyright 2018 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of ISAC. Copyright 2018 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of ISAC.
SponsorsGrant sponsor: Bill and Melinda Gates Foundation Collaboration for AIDS Vaccine Discovery; Grant numbers: OPP1146996, OPP1032144, OPP1032325, and OPP1033098 Grant sponsor: NIH, Grant numbers: R01/R56; AI-52779, 5P30 AI064518 supporting the Duke University Center for AIDS Research (CFAR), and P01 AI120756.
Keywordantibody-dependent cell-mediated cytotoxicity
natural killer cells
peripheral blood mononuclear cells
Identifier to cite or link to this itemhttps://www.scopus.com/inward/record.uri?eid=2-s2.0-85042648926&doi=10.1002%2fcyto.a.23348&partnerID=40&md5=8e619f2cf7f71f61162f990b09df0317; http://hdl.handle.net/10713/9652
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