Related Experiment Video
Updated: Jul 3, 2026
Advances in Human Induced Pluripotent Stem Cell-Derived Chimeric Antigen Receptor-Expressing Natural Killer Cells
Published on: February 14, 2025
Chimeric NKG2D receptor-expressing T cells as an immunotherapy for multiple myeloma
Amorette Barber1, Tong Zhang, Christina J Megli
1Department of Microbiology and Immunology, Dartmouth Medical School, Dartmouth Hitchcock Medical Center, Lebanon, NH 03756, USA.
Objective:
Most myeloma tumor cells from patients express NKG2D ligands. We have reported the development of a chimeric NKG2D receptor (chNKG2D), which consists of the NKG2D receptor fused to the CD3zeta chain. T cells expressing this receptor kill and produce cytokines in response to NKG2D-ligand+ tumor cells. Therefore, we investigated whether human chNKG2D T cells respond against human myeloma cells.
Materials And Methods:
ChNKG2D T cells were generated from healthy donors and myeloma patients. The effector phase of chNKG2D T cells was analyzed by cell-surface marker expression and human myeloma cell lines were tested for expression of NKG2D ligands. Lysis of myeloma cell lines and cytokine secretion by chNKG2D T cells was determined. ChNKG2D T cells grown in serum-free media, or cyropreserved, were assessed for effector cell functions.
Results:
Myeloma cell lines expressed NKG2D ligands. ChNKG2D T cells from healthy donors and myeloma patients lysed myeloma cells, and secreted proinflammatory cytokines when cultured with myeloma cells or patient bone marrow, but not with peripheral blood mononuclear cells or normal bone marrow. Lysis of myeloma cells was dependent on chNKG2D T-cell expression of NKG2D and perforin. Additionally, chNKG2D T cells upregulated CD45RO, did not express CD57, and maintained expression of CD27, CD62L, and CCR7, indicating that the T cells were at an early effector stage. Finally, we showed that chNKG2D T cells generated with serum-free media, or when cryopreserved, maintained effector functions.
Conclusion:
ChNKG2D T cells respond to human myeloma cells and can be generated using clinically applicable cell culture techniques.
Insights
Engineered T cells expressing a chimeric NKG2D receptor (chNKG2D) effectively target and eliminate human myeloma cells. These chNKG2D T cells can be generated using clinically viable methods, offering a promising new avenue for multiple myeloma treatment.
Area of Science:
- Immunology
- Oncology
- Cell Therapy
Background:
- Most multiple myeloma tumor cells express NKG2D ligands.
- Chimeric NKG2D receptor (chNKG2D) technology fuses the NKG2D receptor to the CD3zeta chain.
- T cells expressing chNKG2D can recognize and eliminate NKG2D-ligand-expressing tumor cells.
Purpose of the Study:
- To investigate the efficacy of human chNKG2D T cells against human myeloma cells.
- To determine if chNKG2D T cells can be generated using clinically applicable methods.
Main Methods:
- Generation of chNKG2D T cells from healthy donors and myeloma patients.
- Analysis of NKG2D ligand expression on human myeloma cell lines.
- Assessment of myeloma cell lysis and cytokine secretion by chNKG2D T cells.
- Evaluation of effector cell functions in serum-free media and after cryopreservation.
Main Results:
- Myeloma cell lines express NKG2D ligands.
- chNKG2D T cells from both healthy donors and myeloma patients effectively lysed myeloma cells and secreted proinflammatory cytokines.
- Lysis was dependent on chNKG2D T-cell expression of NKG2D and perforin.
- chNKG2D T cells maintained an early effector phenotype and retained function after serum-free culture or cryopreservation.
Conclusions:
- chNKG2D T cells demonstrate a robust response against human myeloma cells.
- The generation of functional chNKG2D T cells is feasible using clinically applicable cell culture techniques.
