Authors: Berg, Maria¹; Lundqvist, Andreas¹; McCoy, Philip²; Samsel, Leigh²; Fan, Yong³; Tawab, Abdul³; Childs, Richard¹

Source: Cytotherapy, Volume 11, Number 3, May 2009 , pp. 341-355(15)

Publisher: Informa Healthcare

Abstract:

Background aims Cancer immunotherapy involving natural killer (NK) cell infusions and administration of therapeutic agents modulating the susceptibility of tumors to NK-cell lysis has been proposed recently. We provide a method for expanding highly cytotoxic clinical-grade NK cells in vitro for adoptive transfer following bortezomib treatment in patients with advanced malignancies. Methods NK cells were expanded with irradiated Epstein-Barr virus-transformed lymphoblastoid cells. Expanded cells were evaluated for their phenotype, cytotoxicity, cytokine secretion, dependence on interleukin (IL)-2 and ability to retain function after cryopreservation. Results A pure population of clinical-grade NK cells expanded 490±260-fold over 21 days. Expanded NK cells had increased TRAIL, FasL and NKG2D expression and significantly higher cytotoxicity against bortezomib-treated tumors compared with resting NK cells. Expanded NK cells, co-cultured with K562 and renal cell carcinoma tumor targets, secreted significantly higher levels of soluble Fas ligand 6; fgjhd IFN-γ, GM-CSF, TNF-α, MIP-1α and MIP-1β compared with resting NK cells. Secretion of the above cytokines and NK-cell cytolytic function were IL-2 dose dependent. Cryopreservation of expanded NK cells reduced expression of NKG2D and TRAIL and NK-cell cytotoxicity, although this effect could be reversed by exposure of NK cells to IL-2. Conclusions We describe a method for large-scale expansion of NK cells with increased expression of activating receptors and death receptor ligands resulting in superior cytotoxicity against tumor cells. This ex vivo NK-cell expansion technique is currently being utilized in a clinical trial evaluating the anti-tumor activity of adoptively infused NK cells in combination with bortezomib.

Keywords: Bortezomib; expansion; immunotherapy; natural killer cells

Document Type: Research article

DOI: http://dx.doi.org/10.1080/14653240902807034

Affiliations: 1: Hematology Branch, 2: Flow Cytometry Core Facility, National Heart, Lung and Blood Institute, 3: Department of Transfusion Medicine, Cell Processing Section, National Institutes of Health, Bethesda, Maryland, USA

Publication date: 2009-05-01

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