Human and mouse iNKT cell expansion and engineering with viral vectors

Area of Science:

• Immunology
• Cell Therapy
• Oncology

Background:

• Invariant natural killer T (iNKT) cells are crucial for tumor immunosurveillance.
• iNKT cells can be engineered for targeted anti-cancer therapies.
• In vitro expansion is vital for generating sufficient iNKT cells for adoptive cell therapy (ACT).

Purpose of the Study:

• To describe robust methods for isolating and expanding primary mouse and human iNKT cells.
• To enable efficient genetic modification of iNKT cells for tumor-specific targeting.
• To generate large numbers of iNKT cells for pre-clinical studies and potential ACT.

Main Methods:

• Isolation of iNKT cells from mouse spleens and human buffy coats.
• Activation of iNKT cells using anti-CD3/CD28 beads, IL-2, and IL-7.
• Transduction of iNKT cells with tumor-specific chimeric antigen receptors (CARs) or T-cell receptors (TCRs).

Main Results:

• Highly enriched populations of mouse and human iNKT cells were obtained.
• Successful genetic modification and expansion yielded millions of tumor-redirected iNKT cells.
• The resulting iNKT cell product was pure, stable, and suitable for pre-clinical ACT.

Conclusions:

• The described methods provide efficient means to isolate, expand, and genetically engineer iNKT cells.
• These protocols facilitate the generation of iNKT cell-based therapies for hematological and solid malignancies.
• The study supports the advancement of iNKT cell therapy for cancer treatment.