title: Human NK cells activated by EBV+lymphoblastoid cells overcome anti-apoptotic mechanisms of drug resistance in haematological cancer cells.
abstract: Natural killer (NK) cells recognize and eliminate transformed or infected cells that have downregulated MHC class-I and express specific activating ligands. Recent evidence indicates that allogeneic NK cells are useful to eliminate haematological cancer cells independently of MHC-I expression. However, it is unclear if transformed cells expressing mutations that confer anti-apoptotic properties and chemoresistance will be susceptible to NK cells. Allogeneic primary human NK cells were activated using different protocols and prospectively tested for their ability to eliminate diverse mutant haematological and apoptotic-resistant cancer cell lines as well as patient-derived B-cell chronic lymphocytic leukemia cells with chemotherapy multiresistance. Here, we show that human NK cells from healthy donors activatedin vitrowith Epstein Barr virus positive (EBV+)-lymphoblastoid cells display an enhanced cytotoxic and proliferative potential in comparison to other protocols of activation such a K562 cells plus interleukin (IL)2. This enhancement enables them to kill more efficiently a variety of haematological cancer cell lines, including a panel of transfectants that mimic natural mutations leading to oncogenic transformation and chemoresistance (e.g., overexpression of Bcl-2, Bcl-XLand Mcl-1 or downregulation of p53, Bak/Bax or caspase activity). The effect was also observed against blasts from B-cell chronic lymphocytic leukemia patients showing multi-resistance to chemotherapy. Our findings demonstrate that particularin vitroactivated NK cells may overcome anti-apoptotic mechanisms and oncogenic alterations frequently occurring in transformed cells, pointing toward the use of EBV+-lymphoblastoid cells as a desirable strategy to activate NK cellsin vitrofor the purpose of treating haematological neoplasia with poor prognosis.
authors.affiliation: Immune Effector Cells Group (ICE); Aragón Health Research Institute (IIS Aragón); Edificio CIBA; Biomedical Research Center of Aragón (CIBA) ; Zaragoza, Spain ; Cell Immunity in Inflammation; Infection and Cancer Group; Department of Biochemistry and Molecular and Cell Biology; University of Zaragoza ; Zaragoza, Spain.
authors.forename: Diego
authors.initials: D
authors.lastname: Sánchez-Martínez
authors.affiliation: Servicio de Hematología; Hospital Clínico Universitario; Instituto Aragonés de Ciencias de la Salud (IACS); Zaragoza, Spain.
authors.forename: Gemma
authors.initials: G
authors.lastname: Azaceta
authors.affiliation: Immunity and infection Lab; IMIM (Hospital del Mar Medical Research Institute) ; Barcelona, Spain.
authors.forename: Aura
authors.initials: A
authors.lastname: Muntasell
authors.affiliation: Apoptosis; Cancer and Immunity Group; Department of Biochemistry and Molecular and Cellular Biology; University of Zaragoza ; Zaragoza, Spain.
authors.forename: Nacho
authors.initials: N
authors.lastname: Aguiló
authors.affiliation: Immune Effector Cells Group (ICE); Aragón Health Research Institute (IIS Aragón); Edificio CIBA; Biomedical Research Center of Aragón (CIBA) ; Zaragoza, Spain ; Cell Immunity in Inflammation; Infection and Cancer Group; Department of Biochemistry and Molecular and Cell Biology; University of Zaragoza ; Zaragoza, Spain.
authors.forename: David
authors.initials: D
authors.lastname: Núñez
authors.affiliation: Immune Effector Cells Group (ICE); Aragón Health Research Institute (IIS Aragón); Edificio CIBA; Biomedical Research Center of Aragón (CIBA) ; Zaragoza, Spain ; Instituto de Carboquímica ICB-CSIC ; Zaragoza, Spain.
authors.forename: Eva M
authors.initials: EM
authors.lastname: Gálvez
authors.affiliation: Immune Effector Cells Group (ICE); Aragón Health Research Institute (IIS Aragón); Edificio CIBA; Biomedical Research Center of Aragón (CIBA) ; Zaragoza, Spain ; Apoptosis; Cancer and Immunity Group; Department of Biochemistry and Molecular and Cellular Biology; University of Zaragoza ; Zaragoza, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Naval
authors.affiliation: Immune Effector Cells Group (ICE); Aragón Health Research Institute (IIS Aragón); Edificio CIBA; Biomedical Research Center of Aragón (CIBA) ; Zaragoza, Spain ; Apoptosis; Cancer and Immunity Group; Department of Biochemistry and Molecular and Cellular Biology; University of Zaragoza ; Zaragoza, Spain.
authors.forename: Alberto
authors.initials: A
authors.lastname: Anel
authors.affiliation: Servicio de Hematología; Hospital Clínico Universitario; Instituto Aragonés de Ciencias de la Salud (IACS); Zaragoza, Spain.
authors.forename: Luis
authors.initials: L
authors.lastname: Palomera
authors.affiliation: Immunogenetics & HLA; Instituto de Investigación Sanitaria Puerta de Hierro ; Majadahonda, Spain.
authors.forename: Carlos
authors.initials: C
authors.lastname: Vilches
authors.affiliation: Immune Effector Cells Group (ICE); Aragón Health Research Institute (IIS Aragón); Edificio CIBA; Biomedical Research Center of Aragón (CIBA) ; Zaragoza, Spain ; Apoptosis; Cancer and Immunity Group; Department of Biochemistry and Molecular and Cellular Biology; University of Zaragoza ; Zaragoza, Spain.
authors.forename: Isabel
authors.initials: I
authors.lastname: Marzo
authors.affiliation: INSERM, U1040; Université de Montpellier 1; UFR Medecine; Montpellier , France ; Institut de Regenerative Medicine et Biothérapie (IRMB); CHU Montpellier ; Montpellier, France.
authors.forename: Martín
authors.initials: M
authors.lastname: Villalba
authors.affiliation: Immune Effector Cells Group (ICE); Aragón Health Research Institute (IIS Aragón); Edificio CIBA; Biomedical Research Center of Aragón (CIBA) ; Zaragoza, Spain ; Cell Immunity in Inflammation; Infection and Cancer Group; Department of Biochemistry and Molecular and Cell Biology; University of Zaragoza ; Zaragoza, Spain ; Aragón I+D Foundation (ARAID); Government of Aragon , Zaragoza, Spain ; Nanoscience Institute of Aragon (INA); University of Zaragoza , Zaragoza, Spain.
authors.forename: Julián
authors.initials: J
authors.lastname: Pardo
keywords:  B-CLL, B cell chronic lymphocytic leukemia; B lymphoblastoid cell line; EBV, Epstein-Barr virus; IAP, inhibitor of apoptosis; KIR, killer inhibitory receptor; LCL, lymphoblastoid B cell line; NK cells; NK, natural killer; NKR, NK cell receptor; PBL, peripheral blood lymphocyte; PBMC, peripheral blood mononuclear cell; Tc, cytotoxic T; apoptosis; haematological neoplasia; multidrug acquired resistance