NK Cell-Mediated Cytotoxicity Assay

This was performed as previously described (14 (link), 15 (link)). NK cells were labeled with 3 μM of CellTracker™ Violet BMQC Dye (Life Technologies) and incubated overnight with target cells at different E:T ratios. Subsequently, phosphatidylserine (PS) translocation and membrane damage were analyzed in the violet fluorescence-negative target cell population by flow cytometry using Annexin V-FITC (Immunostep) and 7AAD (BD Biosciences) or propidium iodide (PI) as previously described (16 (link), 17 (link)). We consider all cells positive for annexin-V and/or PI (or 7-ADD) as dead (or dying).

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Alexia C., Cren M., Louis-Plence P., Vo D.N., El Ahmadi Y., Dufourcq-Lopez E., Lu Z.Y., Hernandez J., Shamilov F., Chernysheva O., Vasilieva M., Vorotnikov I., Vishnevskay Y., Tupitsyn N., Rossi J.F, & Villalba M. (2019). Polyoxidonium® Activates Cytotoxic Lymphocyte Responses Through Dendritic Cell Maturation: Clinical Effects in Breast Cancer. Frontiers in Immunology, 10, 2693.

Publication 2019

CellTracker™ Violet BMQC Dye

7aad Annexin v Annexin v fitc Cells Flow cytometry Fluorescence Membrane Nk cells Phosphatidylserine Propidium iodide Target population Translocation Violet

Corresponding Organization : Centre National de la Recherche Scientifique

Other organizations : Ministry of Health of the Russian Federation, Voronezh Regional Clinical Oncology Center, Institut Sainte Catherine

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Variable analysis

independent variables

E:T (Effector:Target) ratios

dependent variables

Phosphatidylserine (PS) translocation
Membrane damage

control variables

NK cells labeled with 3 μM of CellTracker™ Violet BMQC Dye (Life Technologies)
Overnight incubation of NK cells with target cells

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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