American Association for Cancer Research
10780432ccr161524-sup-167897_1_supp_3702777_hhzbhh.png (27.25 kB)

Supplementary Figure S3 from Cytokine-Induced Killer Cells Kill Chemo-surviving Melanoma Cancer Stem Cells

Download (27.25 kB)
posted on 2023-03-31, 19:42 authored by Loretta Gammaitoni, Lidia Giraudo, Marco Macagno, Valeria Leuci, Giulia Mesiano, Ramona Rotolo, Francesco Sassi, Martina Sanlorenzo, Alessandro Zaccagna, Alberto Pisacane, Rebecca Senetta, Michela Cangemi, Giulia Cattaneo, Valentina Martin, Valentina Coha, Susanna Gallo, Ymera Pignochino, Anna Sapino, Giovanni Grignani, Fabrizio Carnevale-Schianca, Massimo Aglietta, Dario Sangiolo

Supplementary Figure S3:Expression of NKG2D ligands and PDL1 on putative mCSCs following different treatments.


Associazione Italiana Ricerca sul Cancro

IG grant


Ministero della Salute 2012

Ricerca Finalizzata-Giovani Ricercatori Ministero della Salute

University of Torino Fondo Ricerca Locale 2013

Associazione Italiana Ricerca sul Cancro–AIRC

University of Turin

L'Oréal-UNESCO For Women in Science Fellowship 2016



Purpose: The MHC-unrestricted activity of cytokine-induced killer (CIK) cells against chemo-surviving melanoma cancer stem cells (mCSC) was explored, as CSCs are considered responsible for chemoresistance and relapses.Experimental Design: Putative mCSCs were visualized by engineering patient-derived melanoma cells (MC) with a lentiviral vector encoding eGFP under expression control by stemness gene promoter oct4. Their stemness potential was confirmed in vivo by limiting dilution assays. We explored the sensitivity of eGFP+ mCSCs to chemotherapy (CHT), BRAF inhibitor (BRAFi) or CIK cells, as single agents or in sequence, in vitro. First, we treated MCs in vitro with fotemustine or dabrafenib (BRAF-mutated cases); then, surviving MCs, enriched in mCSCs, were challenged with autologous CIK cells. CIK cell activity against chemoresistant mCSCs was confirmed in vivo in two distinct immunodeficient murine models.Results: We visualized eGFP+ mCSCs (14% ± 2.1%) in 11 MCs. The tumorigenic precursor rate in vivo was higher within eGFP+ MCs (1/42) compared with the eGFP− counterpart (1/4,870). In vitro mCSCs were relatively resistant to CHT and BRAFi, but killed by CIK cells (n = 11, 8/11 autologous), with specific lysis ranging from 95% [effector:tumor ratio (E:T), 40:1] to 20% (E:T 1:3). In vivo infusion of autologous CIK cells into mice bearing xenografts from three distinct melanomas demonstrated significant tumor responses involving CHT-spared eGFP+ mCSCs (P = 0.001). Sequential CHT–immunotherapy treatment retained antitumor activity (n = 12, P = 0.001) reducing mCSC rates (P = 0.01).Conclusions: These findings are the first demonstration that immunotherapy with CIK cells is active against autologous mCSCs surviving CHT or BRAFi. An experimental platform for mCSC study and rationale for CIK cells in melanoma clinical study is provided. Clin Cancer Res; 23(9); 2277–88. ©2016 AACR.

Usage metrics

    Clinical Cancer Research



    Ref. manager