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Supplementary video 1 from Tumor-Produced Interleukin-8 Attracts Human Myeloid-Derived Suppressor Cells and Elicits Extrusion of Neutrophil Extracellular Traps (NETs)

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posted on 2023-03-31, 18:27 authored by Carlos Alfaro, Alvaro Teijeira, Carmen Oñate, Guiomar Pérez, Miguel F. Sanmamed, Maria Pilar Andueza, Diego Alignani, Sara Labiano, Arantza Azpilikueta, Alfonso Rodriguez-Paulete, Saray Garasa, Juan P. Fusco, Angela Aznar, Susana Inogés, Maria De Pizzol, Marcello Allegretti, Jose Medina-Echeverz, Pedro Berraondo, Jose L. Perez-Gracia, Ignacio Melero

Supplementary video 1. IL-8 induces NET extrusion of GrMDSCs using time-lapse in vivo confocal fluorescence microsocopy. GrMDSCs nuclei were pre-stained with DRAQ5 (Red) and treated with IL-8. The non-cell permeant green DNA probe (SYTOXgreen) is present in the culture media and only stains DNA when is extruded extracelullarly. Video represent over five hours a maximum intensity projection of a Zstack. Video is 600x accelerated

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ARTICLE ABSTRACT

Purpose: Myeloid-derived suppressor cells (MDSC) are considered an important T-cell immunosuppressive component in cancer-bearing hosts. The factors that attract these cells to the tumor microenvironment are poorly understood. IL8 (CXCL8) is a potent chemotactic factor for neutrophils and monocytes.Experimental Design: MDSC were characterized and sorted by multicolor flow cytometry on ficoll-gradient isolated blood leucokytes from healthy volunteers (n = 10) and advanced cancer patients (n = 28). In chemotaxis assays, sorted granulocytic and monocytic MDSC were tested in response to recombinant IL8, IL8 derived from cancer cell lines, and patient sera. Neutrophil extracellular traps (NETs) formation was assessed by confocal microscopy, fluorimetry, and time-lapse fluorescence confocal microscopy on short-term MDSC cultures.Results: IL8 chemoattracts both granulocytic (GrMDSC) and monocytic (MoMDSC) human MDSC. Monocytic but not granulocytic MDSC exerted a suppressor activity on the proliferation of autologous T cells isolated from the circulation of cancer patients. IL8 did not modify the T-cell suppressor activity of human MDSC. However, IL8 induced the formation of NETs in the GrMDSC subset.Conclusions: IL8 derived from tumors contributes to the chemotactic recruitment of MDSC and to their functional control. Clin Cancer Res; 22(15); 3924–36. ©2016 AACR.