Artificial feeder cells expressing ligands for killer cell immunoglobulin-like receptors and CD94/NKG2A for expansion of functional primary natural killer cells with tolerance to self

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Susanne Michen - , Department of Neurosurgery, Dresden University of Technology (Author)
  • Jennifer Frosch - , Dresden University of Technology (Author)
  • Monika Füssel - , DKMS Life Science Lab gGmbH (Author)
  • Gabriele Schackert - , Dresden University of Technology, German Cancer Research Center (DKFZ) (Author)
  • Frank Momburg - , German Cancer Research Center (DKFZ) (Author)
  • Achim Temme - , Department of Neurosurgery, Dresden University of Technology, German Cancer Research Center (DKFZ) (Author)

Abstract

Background aims: Natural killer (NK) cells are promising cells for immunotherapy of cancer, and there are ongoing efforts to improve their ex vivo expansion to clinically relevant numbers. This study focused on the development of a C1-, C2-, Bw4 killer cell immunoglobulin-like receptor (KIR) ligand and NKG2A ligand-containing feeder cell line for autonomous expansion of functional NK cells. Methods: PC3PSCA-derived feeder cells expressing IL-2, 4-1BBL and membrane-bound IL-15-mutDAP12 (mIL-15d) fusion protein in combinations or alone were generated and used for expansion. Expanded NK cells were analyzed with respect to subpopulations, expression of NK cell receptors and immune checkpoint molecules as well as their cytotoxicity against K562 cells, cetuximab-marked tumor cells and autologous B cells. Results: Only combinatorial expression of IL-2 plus 4-1BBL or IL-2, 4-1BBL plus mIL-15d in feeder cells efficiently expanded NK cells and supported selective outgrowth of NK cells from peripheral blood mononuclear cell samples. Best expansion of NK cells was achieved using PC3PSCA-IL-2-4-1BBL-mIL-15d feeder cells. Such expanded NK cells exhibited upregulation of natural cytotoxicity receptors, DNAM-1 and NKG2C and induced expression of high affinity IL-2 receptor, which were paralleled by attenuated KIR and increased expression of NKG2A and ILT2. In addition, elevated TIM-3 levels were noted and PD-1 and T cell immunoreceptor with Ig and ITIM domain (TIGIT) levels remained low. Expanded NK cells were highly cytolytic when encountering K562 cells and cetuximab-marked target cells but remained unresponsive to autologous B cells and target cells with protective levels of human leukocyte antigen. Conclusions: Collectively, the results demonstrate the feasibility of PC3PSCA-IL-2-4-1BBL-mIL-15d feeder cells for robust expansion of NK cells, which remain tolerant to self and could be used in the future for adoptive cell therapy of cancer.

Details

Original languageEnglish
Pages (from-to)354-368
Number of pages15
JournalCytotherapy
Volume22
Issue number7
Publication statusPublished - Jul 2020
Peer-reviewedYes

External IDs

PubMed 32451262
ORCID /0000-0002-5726-386X/work/142249125

Keywords

Sustainable Development Goals

Keywords

  • Expansion, Feeder cell line, Natural killer cells, PBMC