Journal: iScience

Article Title: Leveraging Vγ9Vδ2 T cells against prostate cancer through a VHH-based PSMA-Vδ2 bispecific T cell engager

doi: 10.1016/j.isci.2024.111289

Figure Lengend Snippet: Target binding, Vγ9Vδ2 T cell activation, and tumor cell lysis by the PSMA-Vδ2-Fc bsTCE (A) PSMA-Vδ2-Fc bsTCE and control bsTCE binding to expanded Vγ9Vδ2 T cells ( n = 6), LNCaP (left: n = 3, right: n = 2), and PC3 cells ( n = 1). (B) PSMA expression by 22Rv1 and VCaP shown as histograms. (C) Vγ9Vδ2 T cell CD107a expression, CD25 expression, and IFN-γ secretion ( n = 3). (D) Target cell lysis ( n = 3–4) in 24 h co-cultures of expanded Vγ9Vδ2 T cells with 22Rv1, VCaP, LNCaP (WT or PSMA KO), or PC3 with indicated concentrations of bsTCE. (E) Vγ9Vδ2 T cell CD25 expression ( n = 3) and target cell lysis ( n = 3) in 24 h co-cultures of expanded Vγ9Vδ2 T cells with 22Rv1, VCaP, LNCaP, or PC3 with 100 nM PSMA-Vδ2-Fc bsTCE or control bsTCEs. Data were generated using flow cytometry (A left, B, C; CD107a and CD25, D, E), ELISA (A, middle and right) or CBA (C, IFNγ). Data represent mean ± S.E.M.

Article Snippet: LNCaP (PSMA + clone FGC, CRL-1740) and PC3 (PSMA − , CRL-1435) were obtained from American Type Culture Collection (ATCC) and VCaP (PSMA + , 06020201-1VL) and 22Rv1 (PSMA + , 5092802) from the European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Binding Assay, Activation Assay, Lysis, Control, Expressing, Generated, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Leveraging Vγ9Vδ2 T cells against prostate cancer through a VHH-based PSMA-Vδ2 bispecific T cell engager

doi: 10.1016/j.isci.2024.111289

Figure Lengend Snippet: Impact of DNAM-1 and NKG2D receptor interactions in PSMA-Vδ2-Fc bsTCE mediated Vγ9Vδ2 T cell activation and prostate tumor cell lysis (A) CD107a expression (4 h culture, n = 4) and (B) tumor cell lysis (24 h culture, n = 4) from cultures of expanded Vγ9Vδ2 T cells pre-incubated with 10 μg/mL anti-Fc receptor Ab and α-DNAM-1 blocking antibody or IgG1 isotype control with LNCaP, VCaP, or 22Rv1 tumor cells with indicated concentrations of PSMA-Vδ2-Fc bsTCE. (C and D) (C) CD107a expression (4 h culture, n = 4) and (D) tumor cell lysis (24 h culture, n = 4) from cultures of expanded Vγ9Vδ2 T cells pre-incubated with 10 μg/mL anti-Fc receptor Ab and α-NKG2D blocking antibody or IgG1 isotype control with LNCaP, VCaP, or 22Rv1 tumor cells with indicated concentrations of PSMA-Vδ2-Fc bsTCE. (E) CD107a expression on Vγ9Vδ2 T cells derived from malignant prostate tissue ( n = 3) of PCa patients after 4 h co-culture with 0.05 nM PSMA-Vδ2-Fc bsTCE and α-DNAM-1 or α-NKG2D blocking Ab. Data were all generated using flow cytometry and circles represent individual data-points. Box and whisker plots indicate the median, 25th to 75th percentiles and minimum to maximum and bar graphs represent mean ± S.E.M. Paired t test (A, C–E). p < 0.05: ∗, p < 0.01: ∗∗ and p < 0.001: ∗∗∗.

Article Snippet: LNCaP (PSMA + clone FGC, CRL-1740) and PC3 (PSMA − , CRL-1435) were obtained from American Type Culture Collection (ATCC) and VCaP (PSMA + , 06020201-1VL) and 22Rv1 (PSMA + , 5092802) from the European Collection of Authenticated Cell Cultures (ECACC).

Techniques: Activation Assay, Lysis, Expressing, Incubation, Blocking Assay, Control, Derivative Assay, Co-Culture Assay, Generated, Flow Cytometry, Whisker Assay