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

Authors

Maia M.C. Bennett-Boehm, University of Nebraska at Omaha
Anna Mahr, University of Nebraska at OmahaFollow
Alexander Regan, University of Nebraska at OmahaFollow
Sean T. Hartwell, University of Nebraska at Omaha
Isabelle Weber, University of Nebraska at OmahaFollow
Arriana Blackmon, University of Nebraska at OmahaFollow
Cami Bisson, University of Nebraska at OmahaFollow
Angela Ngoc Truong, University of Nebraska at OmahaFollow
Bella Circo, University of Nebraska at OmahaFollow
Jaden Nienhueser, University of Nebraska at OmahaFollow
Donald Rogers, University of Nebraska at OmahaFollow
Nathan Booher, University of Nebraska at OmahaFollow
Nithya Rajagopalan, University of Nebraska at Omaha
Jacob W.S. Martens, University of Nebraska at Omaha
Paul Denton, University of Nebraska at OmahaFollow

Author ORCID Identifier

Denton - https://orcid.org/0000-0003-2458-8147

Document Type

Article

Publication Date

11-28-2023

Publication Title

Heliyon

Volume

9

Abstract

Assays to quantify natural killer (NK) cell killing efficacy have traditionally focused on assessing either direct killing or antibody dependent cell-mediated cytotoxicity (ADCC) independently. Due to the probability that immunotherapeutic interventions affect NK cell-mediated direct killing and NK cell-mediated ADCC differently, we developed an assay with the capacity to measure NK cell-mediated direct killing and ADCC simultaneously with cells from the same human donor. Specifically, this design allows for a single NK cell population to be split into several experimental conditions (e.g., direct killing, ADCC), thus controlling for potential confounders associated with human-to-human variation when assessing immunotherapy impacts. Our Natural Killer cell Simultaneous ADCC and Direct Killing Assay (NK-SADKA) allows researchers to reproducibly quantify both direct killing and ADCC by human NK cells. Furthermore, this optimized experimental design allows for concurrent analysis of the NK cells via flow cytometric immunophenotyping of NK cell populations which will facilitate the identification of relationships between NK cell phenotype and the subsequent killing potential. This assay will be valuable for assessing the broader impact(s) of immunotherapy strategies on both modes of NK cell killing.

Comments

This is an open access article that is licensed under the Creative Commons Attribution NonCommercial NoDerivatives license.

DOI: https://doi.org/10.1016/j.heliyon.2023.e22991

Recommended Citation

Bennett-Boehm, Maia M.C.; Mahr, Anna; Regan, Alexander; Hartwell, Sean T.; Weber, Isabelle; Blackmon, Arriana; Bisson, Cami; Truong, Angela Ngoc; Circo, Bella; Nienhueser, Jaden; Rogers, Donald; Booher, Nathan; Rajagopalan, Nithya; Martens, Jacob W.S.; and Denton, Paul, "Development and implementation of natural killer cell simultaneous ADCC and direct killing assay" (2023). Biology Faculty Publications. 192.
https://digitalcommons.unomaha.edu/biofacpub/192

Creative Commons License

This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.