Antigen scaffolds drive preferential expansion of functional genetically engineered CAR and TCR T cells
(1) Johansen KH (2) Ramanathan K (3) Rodriguez-Pardo C (4) Gjelstrup AK (5) Halldrsdttir HR (6) Voss LF (7) Tvingsholm S (8) Grand LS (9) Dionisio E (10) Skriver AH (11) Frederiksen LP (12) Bj¿rnson M (13) Svensson-Frej M (14) Ormh¿j M (15) Hadrup SR
To specifically expand NY-ESO-1 TCR-T and CD19 CAR-T cells following CRISPR-cas9 knock-inKI to the TRAC locus, Johansen et al. decorated dextran scaffolds with IL-2, IL-21, and either pMHC or CAR antigen, respectively. The scaffold increased TCR/CAR expression and expansion over stimulation with the soluble components. Compared to cytokine-expanded T cells, scaffold-expanded T cells had greater proliferative capacity, cytotoxicity through serial tumor rechallenge in vitro, and superior antitumor efficacy and expansion in vivo. Mixtures of scaffolds displaying several pMHCs expanded pooled TCR-T cells.
Contributed by Alex Najibi
(1) Johansen KH (2) Ramanathan K (3) Rodriguez-Pardo C (4) Gjelstrup AK (5) Halldrsdttir HR (6) Voss LF (7) Tvingsholm S (8) Grand LS (9) Dionisio E (10) Skriver AH (11) Frederiksen LP (12) Bj¿rnson M (13) Svensson-Frej M (14) Ormh¿j M (15) Hadrup SR
To specifically expand NY-ESO-1 TCR-T and CD19 CAR-T cells following CRISPR-cas9 knock-inKI to the TRAC locus, Johansen et al. decorated dextran scaffolds with IL-2, IL-21, and either pMHC or CAR antigen, respectively. The scaffold increased TCR/CAR expression and expansion over stimulation with the soluble components. Compared to cytokine-expanded T cells, scaffold-expanded T cells had greater proliferative capacity, cytotoxicity through serial tumor rechallenge in vitro, and superior antitumor efficacy and expansion in vivo. Mixtures of scaffolds displaying several pMHCs expanded pooled TCR-T cells.
Contributed by Alex Najibi
ABSTRACT: T cells engineered to express recombinant T cell receptors (TCRs) or chimeric antigen receptors (CARs) can induce durable clinical responses in solid and hematological malignancies. However, manufacturing is resource intensive and often yields highly differentiated, exhausted T cell products. To address this, we developed an antigen-scaffold (Ag-scaffold) technology that presents cognate antigen alongside stimulatory cytokines to preferentially expand genetically engineered T cells. We expanded CRISPR-Cas9- and lentivirally engineered TCR T cells with peptide MHC (pMHC)-presenting Ag-scaffolds and anti-CD19 CAR T cells with CD19-presenting Ag-scaffolds. By applying cognate pMHC Ag-scaffolds, we achieved on average 69% (±22%) antigen-specific T cells after 14 days of culture with a cytotoxic, proliferative profile. Ag-scaffold expansion enhanced initial TCR and CAR cytotoxicity; repeated rechallenges showed sustained control. In vivo, Ag-scaffold-expanded CRISPR-Cas9-engineered NY-ESO-1-specific TCR T cells and anti-CD19 CAR T cells also showed improved tumor control in lymphoma and solid tumor xenograft models compared to cells expanded with IL2/7/15.
Author Info:
(1) T Cells and Cancer, Experimental and Translational Immunology Section, Department for Health Technology, Technical University of Denmark, 2800 Kongens Lyngby, Denmark. Electron
ic address: krisha@dtu.dk. (2) T Cells and Cancer, Experimental and Translational Immunology Section, Department for Health Technology, Technical University of Denmark, 2800 Kongens Lyngby, Denmark. (3) T Cells and Cancer, Experimental and Translational Immunology Section, Department for Health Technology, Technical University of Denmark, 2800 Kongens Lyngby, Denmark. (4) T Cells and Cancer, Experimental and Translational Immunology Section, Department for Health Technology, Technical University of Denmark, 2800 Kongens Lyngby, Denmark. (5) T Cells and Cancer, Experimental and Translational Immunology Section, Department for Health Technology, Technical University of Denmark, 2800 Kongens Lyngby, Denmark. (6) T Cells and Cancer, Experimental and Translational Immunology Section, Department for Health Technology, Technical University of Denmark, 2800 Kongens Lyngby, Denmark. (7) T Cells and Cancer, Experimental and Translational Immunology Section, Department for Health Technology, Technical University of Denmark, 2800 Kongens Lyngby, Denmark. (8) T Cells and Cancer, Experimental and Translational Immunology Section, Department for Health Technology, Technical University of Denmark, 2800 Kongens Lyngby, Denmark. (9) T Cells and Cancer, Experimental and Translational Immunology Section, Department for Health Technology, Technical University of Denmark, 2800 Kongens Lyngby, Denmark. (10) T Cells and Cancer, Experimental and Translational Immunology Section, Department for Health Technology, Technical University of Denmark, 2800 Kongens Lyngby, Denmark. (11) T Cells and Cancer, Experimental and Translational Immunology Section, Department for Health Technology, Technical University of Denmark, 2800 Kongens Lyngby, Denmark. (12) T Cells and Cancer, Experimental and Translational Immunology Section, Department for Health Technology, Technical University of Denmark, 2800 Kongens Lyngby, Denmark. (13) T Cells and Cancer, Experimental and Translational Immunology Section, Department for Health Technology, Technical University of Denmark, 2800 Kongens Lyngby, Denmark. (14) T Cells and Cancer, Experimental and Translational Immunology Section, Department for Health Technology, Technical University of Denmark, 2800 Kongens Lyngby, Denmark. (15) T Cells and Cancer, Experimental and Translational Immunology Section, Department for Health Technology, Technical University of Denmark, 2800 Kongens Lyngby, Denmark. Electronic address: sirha@dtu.dk.
Citation: Cell Rep Med 2026 Nov 5 103128 Epub11/05/2026
