Adoptive Transfer of Gene-Modified Primary NK Cells Can Specifically Inhibit Tumor Progression In Vivo

Abstract

NK cells hold great potential for improving the immunotherapy of cancer. Nevertheless, tumor cells can effectively escape NK cell-mediated apoptosis through interaction of MHC molecules with NK cell inhibitory receptors. Thus, to harness NK cell effector function against tumors, we used Amaxa gene transfer technology to gene-modify primary mouse NK cells with a chimeric single-chain variable fragment (scFv) receptor specific for the human erbB2 tumor-associated Ag. The chimeric receptor was composed of the extracellular scFv anti-erbB2 Ab linked to the transmembrane and cytoplasmic CD28 and TCR-ζ signaling domains (scFv-CD28-ζ). In this study we demonstrated that mouse NK cells gene-modified with this chimera could specifically mediate enhanced killing of an erbB2+ MHC class I+ lymphoma in a perforin-dependent manner. Expression of the chimera did not interfere with NK cell-mediated cytotoxicity mediated by endogenous NK receptors. Furthermore, adoptive transfer of gene-modified NK cells significantly enhanced the survival of RAG mice bearing established i.p. RMA-erbB2+ lymphoma. In summary, these data suggest that use of genetically modified NK cells could broaden the scope of cancer immunotherapy for patients.

Specific growth inhibition of ErbB2‑expressing human breast cancer cells by genetically modified NK‑92 cells

“…IFN-γ is a pleiotropic cytokine capable of influencing both the innate and adaptive immune systems (34). The production of IFN-γ by active NK cells is critical for IL-12 antitumor therapy (35). Rapid NK cell activation is linked to the activation of professional antigen presenting cells (APCs), which produce IL-12.…”

Rapid development of exhaustion and down-regulation of eomesodermin limit the antitumor activity of adoptively transferred murine natural killer cells

“…We acknowledge that these findings are at some variance with previously published results. [6][7][8][9] However we attempted to model our experiments on clinically relevant settings including the infusion of no more than clinically achievable NK-cell numbers (2 ϫ 10 7 /kg in clinical reports, 36 5 ϫ 10 7 /kg in our experiments) with the administration of NK cells occurring at a site and time distant to tumor implantation.…”

“…5 Although prolongation of survival after adoptive NK therapy has been shown to occur in several mouse models, long-term disease-free survival is rare despite experimental conditions including the administration of higher doses of NK cells than are clinically feasible, colocalized injection of tumor with NK cells, depletion of regulatory T cells with additional immunomodulatory therapy, or genetic modification of the NK cells. [6][7][8][9] To delineate the barriers to successful NK immunotherapy, we traced the fate of freshly isolated adoptively transferred NK cells using several murine tumor models. We found that NK cells rapidly home to and accumulate within tumor sites, yet fail to reject the tumor because of a rapid down-regulation of activating receptors and deactivation of effector functions, such as cytotoxicity and cytokine production.…”

ESAT-6–dependent cytosolic pattern recognition drives noncognate tuberculosis control in vivo

“…NK cells were negatively isolated from the spleens and LNs of donor mice using an NK cell enrichment kit and MACS Cell Separation Technology (Miltenyi Biotec) according to the manufacturer's instructions. Isolated and purified NK cells were cultured with recombinant human IL-2 (Peprotech) as described previously (85). The purity of cells after culture was determined by flow cytometry, and 1 × 10 6 NK cells were adoptively transferred i.v.…”