Human CD3 complex is required for the generation of T-cell receptor-like chimeric antigen receptor targeting WT1 in natural killer cells
Issued Date
2026-04-01
Resource Type
ISSN
03407004
eISSN
14320851
Scopus ID
2-s2.0-105033500221
Journal Title
Cancer Immunology Immunotherapy
Volume
75
Issue
4
Rights Holder(s)
SCOPUS
Bibliographic Citation
Cancer Immunology Immunotherapy Vol.75 No.4 (2026)
Suggested Citation
Luanpitpong S., Poohadsuan J., Samart P., Kang X., Pratumkaew P., Janan M., Issaragrisil S. Human CD3 complex is required for the generation of T-cell receptor-like chimeric antigen receptor targeting WT1 in natural killer cells. Cancer Immunology Immunotherapy Vol.75 No.4 (2026). doi:10.1007/s00262-026-04352-9 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/115932
Title
Human CD3 complex is required for the generation of T-cell receptor-like chimeric antigen receptor targeting WT1 in natural killer cells
Author's Affiliation
Corresponding Author(s)
Other Contributor(s)
Abstract
Chimeric antigen receptor-natural killer (CAR-NK) cells represent a promising cellular immunotherapy platform for various cancers, offering a favorable clinical safety profile and potential to generate off-the-shelf products. However, broader application of CAR-NK cells is in part restricted by the availability of surface tumor antigens. T-cell receptor (TCR)-like CARs have gained increasing attention due to their ability to recognize peptides derived from intracellular tumor antigens presented by major histocompatibility complex molecules. In the present study, we designed a third-generation CAR targeting WT1(126–134) peptide presented by HLA-A*02:01, harboring a TCR-like scFv linked to CD28, 4-1BB, and CD3ζ signaling domains (CAR-WT1), and introduced it into human NK-92 cells—the only FDA-approved NK cell line for clinical trials. Interestingly, we found that surface expression of CAR-WT1 in NK-92 cells required the presence of human CD3 complex, as observed for full-length TCRs. Quantitative PCR and subsequent pathway and network analyses indicated enhanced immune activation potentially relevant to effector function in CAR-WT1/CD3 NK-92 cells. The established CAR-WT1/CD3 NK-92 cells exhibited significantly greater cytotoxicity against WT1<sup>+</sup>/HLA-A2<sup>+</sup> target tumor cells than its non-transduced NK-92 cells, consistent with the release of functional cytokines. Once CAR-WT1 was expressed on NK-92 cell surface, surface CD3 appeared dispensable for NK cytotoxicity. Together, our findings provide evidence supporting the feasibility for generating functional TCR-like CAR-WT1 NK-92 cells, thereby broadening the scope of targetable antigens for CAR-NK cell therapy to include intracellular antigens. Other TCR-like CARs with impaired surface expression may be introduced in NK-92 cells using the same strategy, pending further validation.