1. A flow cytometry based assay that simultaneously measures cytotoxicity and monocyte mediated antibody dependent effector activity
- Author
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Aljawharah Alrubayyi, Dominic Paquin-Proulx, Kristopher M. Paolino, Michael A. Eller, Merlin L. Robb, Mark de Souza, Julie A Ake, Nelson L. Michael, Alexandra Schuetz, Kerri G. Lal, and Surat Jongrakthaitae
- Subjects
Male ,0301 basic medicine ,Phagocytosis ,Immunology ,Monocytes ,Flow cytometry ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Humans ,Immunology and Allergy ,Organic Chemicals ,Cytotoxicity ,Antibody-dependent cell-mediated cytotoxicity ,biology ,medicine.diagnostic_test ,Chemistry ,Effector ,Monocyte ,Antibody-Dependent Cell Cytotoxicity ,Flow Cytometry ,Molecular biology ,Killer Cells, Natural ,030104 developmental biology ,Cell killing ,medicine.anatomical_structure ,biology.protein ,Female ,Antibody ,030215 immunology - Abstract
Antibody effector functions such as antibody dependent cellular cytotoxicity (ADCC) and antibody dependent cellular phagocytosis (ADCP) are considered important immunologic parameters following results from the RV144 clinical trial where a reduced risk of infection was associated with non-neutralizing antibody against the V1/V2 region of HIV envelope. The rapid and fluorometric ADCC (RFADCC) assay has been widely used to measure ADCC, however, the mechanism behind the activity measured remains unclear. Here, we demonstrate that monocytes acquire the PKH26 dye used in the RFADCC assay and that the commonly used RFADCC readout correlates with phagocytosis. The RFADCC assay was combined with an amine reactive dye staining to confirm target cell killing. Interestingly, the majority of RFADCC and amine indices were mutually exclusive. In fact, the amine reactive assay results correlated with results from another assays that directly measure NK cell antibody effector functions not associated with phagocytosis. Together, this combined assay offers the opportunity to discriminate monocytes and NK cell antibody effector functions simultaneously.
- Published
- 2018