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Updated: May 20, 2026
A Real-time Potency Assay for Chimeric Antigen Receptor T Cells Targeting Solid and Hematological Cancer Cells
Published on: November 12, 2019
An NKp30-based chimeric antigen receptor promotes T cell effector functions and antitumor efficacy in vivo
Tong Zhang1, Ming-Ru Wu, Charles L Sentman
1Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Lebanon, NH 03756, USA.
Abstract:
NKp30 is a natural cytotoxicity receptor that is expressed on NK cells and recognizes B7-H6, which is expressed on several types of tumors but few normal cells. To target effector T cells against B7-H6+ tumors, we developed several chimeric AgRs (CARs) based on NKp30, which contain the CD28- and/or CD3ζ-signaling domains with the transmembrane domains from CD3ζ, CD28, or CD8α. The data show that chimeric NKp30-expressing T cells responded to B7-H6+ tumor cells. The NKp30 CAR-expressing T cells produced IFN-γ and killed B7-H6 ligand-expressing tumor cells; this response was dependent upon ligand expression on target cells but not on MHC expression. PBMC-derived dendritic cells also express NKp30 ligands, including immature dendritic cells, and they can stimulate NKp30 CAR-bearing T cells to produce IFN-γ, but to a lesser extent. The addition of a CD28-signaling domain significantly enhanced the activity of the NKp30 CAR in a PI3K-dependent manner. Adoptive transfer of T cells expressing a chimeric NKp30 receptor containing a CD28-signaling domain inhibited the growth of a B7-H6-expressing murine lymphoma (RMA/B7-H6) in vivo. Moreover, mice that remained tumor-free were resistant to a subsequent challenge with the wild-type RMA tumor cells, suggesting the generation of immunity against other tumor Ags. Overall, this study demonstrates the specificity and therapeutic potential of adoptive immunotherapy with NKp30 CAR-expressing T cells against B7-H6+ tumor cells in vivo.
Insights
Engineered T cells targeting B7-H6+ tumors show promise. Chimeric NKp30 receptor (CAR) T cells effectively kill cancer cells and inhibit tumor growth in vivo, suggesting a new immunotherapy approach.
Area of Science:
- Immunology
- Oncology
- Cell Therapy
Background:
- NKp30 is a natural cytotoxicity receptor found on NK cells.
- B7-H6 is a ligand recognized by NKp30, expressed on various tumors but rarely on normal tissues.
- This presents a potential target for cancer immunotherapy.
Purpose of the Study:
- To develop and evaluate chimeric antigen receptors (CARs) based on NKp30 for targeting B7-H6-expressing tumors.
- To assess the efficacy of NKp30 CAR-expressing T cells in preclinical models.
Main Methods:
- Engineered chimeric antigen receptors (CARs) incorporating NKp30 domains with CD28 and/or CD3ζ signaling components.
- Testing NKp30 CAR T-cell activity against B7-H6+ tumor cells in vitro, including cytokine production and cytotoxicity assays.
- Evaluating the therapeutic effect of adoptive transfer of NKp30 CAR T cells in a murine lymphoma model.
Main Results:
- NKp30 CAR T cells demonstrated specific recognition and killing of B7-H6+ tumor cells, independent of MHC expression.
- The addition of a CD28 signaling domain enhanced CAR T-cell activity via PI3K signaling.
- Adoptive transfer of NKp30 CAR T cells significantly inhibited tumor growth in vivo and induced long-term immunity.
Conclusions:
- Chimeric NKp30 CAR-expressing T cells are a viable strategy for targeting B7-H6+ tumors.
- This approach shows therapeutic potential for adoptive immunotherapy against specific cancers.
- The study suggests the generation of broader anti-tumor immunity following treatment.
