Bioengineering and serum free expansion of blood-derived γδ T cells.
Cellular immunotherapy relies on several highly variable patient-specific parameters, such as (i) cell number before and after expansion, (ii) targeting of cells to tumors, (iii) cell survival and function after infusion, and (iv) on- and off-target adverse events. Cellular approaches such as the specific expansion of γδ T cells as opposed to αβ T cells are being pursued. γδ T cells are reasonable candidates for immunotherapy because they (i) possess intrinsic anti-tumorigenicity, (ii) require no priming, (iii) direct tumor killing via recognition of stress-responsive ligands, and (iv), as we now show, can be expanded to clinical cell doses in current Good Manufacturing Practice serum-free media (SFM).



γδ T-cell expansion was evaluated in several SFMs. Additionally, the expanded γδ T cells were evaluated for their transduction efficiency using lentiviral vectors (LV).



Of the SFM cultures, robust expansion was only observed in OpTmizer supplemented with high-dose interleukin-2. γδ T-cell percentages and numbers were sufficient for clinical use. Using cells from several donors, transduction efficiencies ranged from 13 to 33%, which is similar to transduction levels observed using αβ T cells with similar multiplicity of infection.



An optimized method of γδT-cell expansion and transduction was developed that can be tested in early-phase clinical trials. With appropriate elimination of the αβT cell-component, the absence of MHC-restriction affords the opportunity for use in the allogeneic setting with limited risk of graft versus host disease. Finally, the use of SFM provides clinically safer, widely applicable and potentially more efficacious cellular immunotherapy.
Aflac Cancer and Blood Disorders Center, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, USA.
Kathryn S
KS
Sutton
Aflac Cancer and Blood Disorders Center, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, USA.
Anindya
A
Dasgupta
Aflac Cancer and Blood Disorders Center, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, USA.
David
D
McCarty
Aflac Cancer and Blood Disorders Center, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, USA.
Christopher B
CB
Doering
Aflac Cancer and Blood Disorders Center, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, USA. Electronic address: hspence@emory.edu.
H Trent
HT
Spencer
D057005:Bioengineering / Q000379:methods*; D016923:Cell Death / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D016895:Culture Media, Serum-Free; D004164:Diphosphonates / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000378:metabolism; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D007167:Immunotherapy; D007376:Interleukin-2 / Q000494:pharmacology; D020014:K562 Cells; D018655:Lymphocyte Count; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000378:metabolism*; D011973:Receptors, LDL / Q000378:metabolism; D013601:T-Lymphocytes / Q000166:cytology* / Q000187:drug effects; D014019:Tissue Donors; D000077211:Zoledronic Acid
D016895:Culture Media, Serum-Free; D004164:Diphosphonates; D007093:Imidazoles; D007376:Interleukin-2; D016692:Receptors, Antigen, T-Cell, gamma-delta; D011973:Receptors, LDL; D000077211:Zoledronic Acid
immunotherapy; lentivector; serum-free media; γδT cells