Development and implementation of natural killer cell simultaneous ADCC and direct killing assay

Bennett-Boehm, Maia M.C.; Mahr, Anna R.; Hartwell, Sean T.; Regan, Alexander K.; Weber, Isabelle S.; Blackmon, Arriana; Bisson, Cami R.; Truong, Angela N.; Circo, Bella A.; Nienhueser, Jaden; Rogers, Donald R.; Booher, Nathan; Rajagopalan, Nithya; Martens, Jacob W.S.; Denton, Paul W.

doi:10.1016/j.heliyon.2023.e22991

Heliyon

2023

DOI: 10.1016/j.heliyon.2023.e22991

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Development and implementation of natural killer cell simultaneous ADCC and direct killing assay

Maia M.C. Bennett-Boehm,

Anna R. Mahr,

Sean T. Hartwell

et al.

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2024

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TLR9 agonism differentially impacts human NK cell-mediated direct killing and antibody-dependent cell-mediated cytotoxicity

Mahr,

Bennett-Boehm,

Rothemejer

et al. 2024

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There are two known mechanisms by which natural killer (NK) cells recognize and kill diseased targets: (i) direct killing and (ii) antibody-dependent cell-mediated cytotoxicity (ADCC). We investigated an indirect NK cell activation strategy for the enhancement of human NK cell killing function. We did this by leveraging the fact that toll-like receptor 9 (TLR9) agonism within pools of human peripheral blood mononuclear cells (PBMCs) results in a robust interferon signaling cascade that leads to NK cell activation. After TLR9 agonist stimulation, NK cells were enriched and incorporated into assays to assess their ability to kill tumor cell line targets. Notably, differential impacts of TLR9 agonism were observed—direct killing was enhanced while ADCC was not increased. To ensure that the observed differential effects were not attributable to differences between human donors, we recapitulated the observation using our Natural Killer—Simultaneous ADCC and Direct Killing Assay (NK-SADKA) that controls for human-to-human differences. Next, we observed a treatment-induced decrease in NK cell surface CD16—known to be shed by NK cells post-activation. Given the essential role of CD16 in ADCC, such shedding could account for the observed differential impact of TLR9 agonism on NK cell-mediated killing capacity.

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TLR9 agonism differentially impacts human NK cell-mediated direct killing and antibody-dependent cell-mediated cytotoxicity

Mahr,

Bennett-Boehm,

Rothemejer

et al. 2024

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

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Development and implementation of natural killer cell simultaneous ADCC and direct killing assay

Cited by 1 publication

References 25 publications

TLR9 agonism differentially impacts human NK cell-mediated direct killing and antibody-dependent cell-mediated cytotoxicity

TLR9 agonism differentially impacts human NK cell-mediated direct killing and antibody-dependent cell-mediated cytotoxicity

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