Application of area scaling analysis to identify natural killer cell and monocyte involvement in the GranToxiLux antibody dependent cell-mediated cytotoxicity assay.

Journal Article (Journal Article)

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.

Full Text

Duke Authors

Cited Authors

  • Pollara, J; Orlandi, C; Beck, C; Edwards, RW; Hu, Y; Liu, S; Wang, S; Koup, RA; Denny, TN; Lu, S; Tomaras, GD; DeVico, A; Lewis, GK; Ferrari, G

Published Date

  • April 2018

Published In

Volume / Issue

  • 93 / 4

Start / End Page

  • 436 - 447

PubMed ID

  • 29498807

Pubmed Central ID

  • PMC5969088

Electronic International Standard Serial Number (EISSN)

  • 1552-4930

Digital Object Identifier (DOI)

  • 10.1002/cyto.a.23348


  • eng

Conference Location

  • United States