In preclinical pancreatic cancer models, Allo15MCAR-NKT cells infiltrate deep into tumors, show strong activation with CD69, IFN-γ, perforin, and granzyme B, experience reduced exhaustion, and achieve superior tumor control with extended survival compared to CAR-T cells, including efficient trafficking to distant sites such as the lungs.

In vitro, Allo15MCAR-NKT cells kill both mesothelin-positive and -negative pancreatic cancer cells, mitigate tumor heterogeneity, sustain cytotoxicity over serial challenges, and rely on NK receptor signaling for killing; conventional CAR-Ts fail against mesothelin-negative or heterogeneous tumors.

A highly pure CAR+/iNKT+ population (>97%) is produced via a 6-week feeder-free differentiation from a single cord blood donor, yielding more than 10^9 therapeutic cells suitable for thousands of doses.

Safety and immunogenicity are favorable: low HLA expression after inflammatory stimuli, minimal alloreactivity, no graft-versus-host disease signs in mice, no weight loss or systemic toxicity, and reduced inflammatory toxicity (lower IL-6 and SAA-3) in a peritoneal CRS model, while preserving antitumor activity.

The engineered cells combine a mesothelin-targeting third-generation CAR, an invariant NKT TCR, and soluble IL-15 to boost persistence, trafficking, and effector function, enabling scalable, off-the-shelf production.

Overall, the study outlines a promising preclinical platform for scalable, universal, off-the-shelf immunotherapy against both primary and metastatic pancreatic cancer, potentially overcoming antigen heterogeneity, tumor microenvironment barriers, and immune escape.

Antigen-independent cytotoxicity was observed, with control of mesothelin-negative tumors and localized tumor activity, indicating resistance to antigen escape common in solid tumors.

Allo15MCAR-NKT cells display a hybrid T/NK phenotype with robust activating receptors (NKG2D, DNAM-1, NKp30, NKp44) and low exhaustion markers (PD-1, TIM-3, LAG-3).

Allogeneic stem-cell–engineered CAR-NKT cells derived from cord blood HSPCs were developed to treat pancreatic cancer, addressing autologous CAR-T manufacturing bottlenecks and tumor resistance.