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

dc.contributor.author Pollara, Justin
dc.contributor.author Denny, Thomas
dc.contributor.author Ferrari, Guido
dc.contributor.author Tomaras, Georgia
dc.contributor.author Anderson, Lettie
dc.contributor.author Shen, Xiaoying
dc.contributor.author Orlandi, Chiara
dc.contributor.author Beck, Charles
dc.contributor.author Edwards, R Whitney
dc.contributor.author Hu, Yi
dc.contributor.author Liu, Shuying
dc.contributor.author Wang, Shixia
dc.contributor.author Koup, Richard A
dc.contributor.author Lu, Shan
dc.contributor.author DeVico, Anthony
dc.contributor.author Lewis, George K
dc.date.accessioned 2018-12-20T16:17:33Z
dc.date.available 2018-12-20T16:17:33Z
dc.date.issued 2018-04
dc.identifier.issn 1552-4922
dc.identifier.issn 1552-4930
dc.identifier.uri https://hdl.handle.net/10161/17800
dc.description.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.
dc.language eng
dc.publisher Wiley
dc.relation.ispartof Cytometry. Part A : the journal of the International Society for Analytical Cytology
dc.relation.isversionof 10.1002/cyto.a.23348
dc.subject GranToxiLux assay
dc.subject HIV
dc.subject antibody-dependent cell-mediated cytotoxicity
dc.subject monocytes
dc.subject natural killer cells
dc.subject peripheral blood mononuclear cells
dc.title Application of area scaling analysis to identify natural killer cell and monocyte involvement in the GranToxiLux antibody dependent cell-mediated cytotoxicity assay.
dc.type Journal article
dc.date.updated 2018-12-20T16:17:30Z
pubs.begin-page 436
pubs.end-page 447
pubs.issue 4
pubs.organisational-group School of Medicine
pubs.organisational-group Duke
pubs.organisational-group Duke Human Vaccine Institute
pubs.organisational-group Institutes and Centers
pubs.organisational-group Surgery, Surgical Sciences
pubs.organisational-group Surgery
pubs.organisational-group Clinical Science Departments
pubs.organisational-group Medicine, Duke Human Vaccine Institute
pubs.organisational-group Medicine
pubs.organisational-group Molecular Genetics and Microbiology
pubs.organisational-group Basic Science Departments
pubs.organisational-group Duke Global Health Institute
pubs.organisational-group University Institutes and Centers
pubs.organisational-group Institutes and Provost's Academic Units
pubs.organisational-group Immunology
pubs.organisational-group Staff
pubs.publication-status Published
pubs.volume 93
duke.contributor.orcid Ferrari, Guido|0000-0001-7747-3349
duke.contributor.orcid Tomaras, Georgia|0000-0001-8076-1931
duke.contributor.orcid Shen, Xiaoying|0000-0001-8076-1931


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