Large Scale Ex Vivo Expansion of γδ T cells Using Artificial Antigen-presenting Cells

Area of Science:

• Immunology
• Cell Therapy
• Cancer Research

Background:

• Gamma delta (γδ) T cells are crucial for anti-cancer immunity but are rare and impaired in patients.
• Existing methods for in vivo expansion of γδ T cells show promise but have limitations.

Purpose of the Study:

• To develop an efficient method for large-scale, ex vivo expansion of allogeneic donor-derived γδ T cells.
• To achieve clinically relevant cell doses using a Good Manufacturing Practice (GMP)-compliant protocol.

Main Methods:

• Utilized a novel quadruplet artificial antigen-presenting cell (aAPC) engineered with K-562 CD3scFv/CD137L/CD28scFv/IL15RA.
• Combined aAPC with zoledronic acid (zol) and interleukin-2 (IL-2) for γδ T cell expansion.
• Employed a GMP-compliant protocol for ex vivo cell culture.

Main Results:

• Achieved a 633-fold expansion of γδ T cells after 10 days of co-culture with aAPC.
• Expanded γδ T cells displayed central (47%) and effector (43%) memory phenotypes.
• >90% of expanded γδ T cells expressed NKG2D with low PD1 and LAG3 expression, demonstrating potent cytotoxic activity against target cells.

Conclusions:

• The developed quadruplet aAPC combined with zol/IL-2 enables efficient, large-scale ex vivo expansion of donor-derived γδ T cells.
• This GMP-compliant method provides sufficient cell doses for future clinical applications in cancer immunotherapy.
• Expanded γδ T cells are functionally potent and suitable for therapeutic use.