Differential effects on natural killer cell production by membrane-bound cytokine stimulations.
Meiping ChangXiaoyan TangLuke NelsonGregg NybergZhimei DuPublished in: Biotechnology and bioengineering (2022)
Robust manufacturing production of natural killer (NK) cells has been challenging in allogeneic NK cell-based therapy. Here, we compared the impact of cytokines on NK cell expansion by developing recombinant K562 feeder cell lines expressing membrane-bound cytokines, mIL15, mIL21, and 41BBL, individually or in combination. We found that 41BBL played a dominant role in promoting up to 500,000-fold of NK cell expansion after a 21-day culture process without inducing exhaustion. However, 41BBL stimulation reduced the overall cytotoxic activity of NK cells when combined with mIL15 and/or mIL21. Additionally, long-term stimulation with mIL15 and/or mIL21, but not 41BBL, increased CD56 expression and the CD56 brigh t population, which is unexpectedly correlated with NK cell cytotoxicity. By conducting single-cell sequencing, we identified distinct subpopulations of NK cells induced by different cytokines, including an adaptive-like CD56 bright CD16 - CD49a + subset induced by mIL15. Through gene expression analysis, we found that different cytokines modulated signaling pathways and target genes involved in cell cycle, senescence, self-renewal, migration, and maturation in different ways. Together, our study demonstrates that cytokine signaling pathways play distinct roles in NK cell expansion and differentiation, which sheds light on NK cell process designs to improve productivity and product quality.
Keyphrases
- nk cells
- metal organic framework
- solid phase extraction
- single cell
- cell cycle
- signaling pathway
- cell proliferation
- rna seq
- poor prognosis
- epithelial mesenchymal transition
- stem cell transplantation
- simultaneous determination
- climate change
- stem cells
- pi k akt
- liquid chromatography
- bone marrow
- endothelial cells
- long non coding rna
- induced apoptosis
- high resolution
- mesenchymal stem cells
- gene expression
- genome wide identification
- high dose
- transcription factor
- low dose
- endoplasmic reticulum stress
- copy number
- binding protein