Mobile website
Basic Research
Effects of target cell overexpression of IL-15, 4-1BBL and IL-18 combine with IL-2 on NK cell activation and cytotoxicity during ex vivo expansion
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
Abstract:
Objective: To evaluate the effects of K562 cells overexpression of IL-15, 4-1BBL and IL-18 combined with IL-2 ex vivo expansion on NK cell activation and cytotoxicity. Methods: K562 cells were engineered to overexpress IL-15, 4-1BBL and IL-18 on the cell membrane. Peripheral blood mononuclear cells (PBMCs) were prepared from venous blood collected from both healthy volunteers and cancer patients. NK cells were purified from PBMCs and expanded ex vivo by co-culture with engineered K562 cells in the presence of IL-2 and/or Hsp70 peptide (TKD) for 21 days. The purity and surface marker expression of the expanded NK cells were analyzed by flow cytometry. The cytotoxicity and antibody-dependent cell-mediated cytotoxicity (ADCC) of the expanded NK cells were assessed by a Cell Counting Ki-8-based colorimetric assay. Results: In the cell preparations from healthy subjects, ex vivo expansion for 3 weeks by co-culture with engineered K562 cells in the presence of IL-2 and/or TKD increased the purity of NK cells to (93±3)%, increased the proportion of cells expressing killer activation receptors, NKG2D, CD94, NKp46, NKp30, and NKp44, by 60%, 40%, 20%, 40%, and 63% respectively with no effect on the proportion of cells expressing inhibitory receptors (CD158b, NKB1 and NKAT2), increased the ADCC by 32%, and increased the cytotoxicity against K562, A549, 7721, MCF tumor cells by 19%, 29%, 26% and 28% respectively. In the cell preparations from cancer patients, ex vivo expansion induce the percentage of NK cells increase to (90.0±8.0)% and the cytotoxicity against K562 cells by 17%.Conclusion: Co-culture with engineered K562 cells overexpressing IL-15, 4-1BBL and IL-18 in the presence of IL-2 and/or TKD may offer an effective approach to expand the NK cell population ex vivo and enhance the NK cell cytotoxicity.
Keywords:
Project Supported:
Project supported by the National Natural Science Foundation of Shaanxi Province（No. 2014JM4171）
Clc Number:
