Journal: Blood Advances

Article Title: TCRVβ-targeting antibody-drug conjugates as a novel strategy to eliminate malignant T cells in T cell cancers

doi: 10.1182/bloodadvances.2026020282

Figure Lengend Snippet: Anti-TCR Vβ2-PNU ADC inhibits tumor growth in vivo. (A) Representative flow cytometric plot showing percentage of viable MOLT-16 (T-ALL) cells after treatment for 72 hours with 1 μg/mL of either anti-TCR Vβ2-PNU or hIgG1-PNU or an unconjugated anti-TCR Vβ2 antibody. (B) Graph showing percentage of viable MOLT-16 (T-ALL) cells after treatment for 72 hours with 1 μg/mL of either anti-TCR Vβ2-PNU or hIgG1-PNU or an unconjugated anti-TCR Vβ2 antibody. n = 3 biological replicates. Ordinary 1-way ANOVA was performed, and data are shown as mean ± s.e.m.; P < .001. (C) Schematic showing timeline of mouse experiment. Treatments were administered IV. Figure created with biorender.com . Vadivel, C. K. (2026) https://BioRender.com/csk02dd . (D) NSG mice were injected subcutaneously with MOLT-16 cells (day 0). Mice were randomized into 4 treatment groups: PBS (n = 7), anti-TCR Vβ2 antibody (n = 8), hIgG1-PNU (n = 8), or anti-TCR Vβ2-PNU ADC (n = 8), all dosed at 0.75 mg/kg. Treatments were administered as IV injections in the tail on day 7, day 14, and day 21. Representative images show tumor growth in the treatment groups (white-dotted circles), except treatment with anti-TCR Vβ2-PNU ADC (black-dotted region). (E) Kaplan-Meier survival curves of MOLT-16–bearing NSG mice, n = 8 (TCR Vβ2-PNU, hIgG1-PNU, anti-TCR Vβ2 antibody) and n = 7 for PBS. The median survival was as follows: 20 days (PBS), 19 days (anti-TCR Vβ2 antibody), 18 days (hIgG1-PNU), and end point not reached (undefined; anti-TCR Vβ2-PNU ADC). Statistical analysis was performed using the log-rank Mantel-Cox test; P < .0001. (F) Cells were extracted from tumors of MOLT-16–bearing NSG mice. Flow cytometric plots showing percentage of viable extracted tumor cells after treatment for 72 hours with either anti-TCR Vβ2-PNU or mIgG1-PNU or an unconjugated anti-TCR Vβ2 antibody at the indicated concentration. ANOVA, analysis of variance; s.e.m., standard error of the mean; T-ALL, T-cell acute lymphoblastic leukemia.

Article Snippet: We purchased anti-TCR Vβ2 custom antibody (REA654/MPB2D5) from Miltenyi Biotec.

Techniques: In Vivo, Injection, Concentration Assay

Journal: Blood Advances

Article Title: TCRVβ-targeting antibody-drug conjugates as a novel strategy to eliminate malignant T cells in T cell cancers

doi: 10.1182/bloodadvances.2026020282

Figure Lengend Snippet: Generated anti-TCR Vβ2-PNU ADC kills malignant T cells in vitro. (A) Schematic overview of the generation of anti-TCR Vβ2-PNU ADC (detailed in “Methods”). The cytotoxic drug PNU-159682 conjugated to the antibody is highlighted in yellow. Chemical structure created with molview.org PubChem Identifier: CID 145712370. Figure created with biorender.com . Vadivel, C. K. (2026) ( https://BioRender.com/2nax8xf ). (B) Efficiency of the PNU conjugation was assessed on a Coomassie-stained SDS-PAGE gel. The complete upward shift in MW of the heavy chain (∼50 kDa) of the ADCs (IgG1-PNU and TCR Vβ2-PNU) compared to the azide-activated antibodies (IgG1-az and TCR Vβ2-az) confirms that both ADCs are homogeneous, with a DAR of 2. (C) Flow cytometric plot showing percentage of CD3 + CD4 + TCR Vβ2 + malignant cells in PBMCs from a patient with leukemic CTCL (L-CTCL). (D) Flow cytometric plot showing viable malignant cells from a patient with TCR Vβ2 + L-CTCL after treatment with either anti-TCR Vβ2-PNU or IgG1-PNU or an unconjugated anti-TCR Vβ2 antibody at the indicated concentration for 72 hours. (E) Representative flow cytometric plots showing the percentage of viable malignant cells in an additional patient with TCR Vβ2 + L-CTCL after treatment with 2 μg/mL of either anti-TCR Vβ2-PNU or hIgG1-PNU or an unconjugated anti-TCR Vβ2 antibody for 72 hours. (F) Graph showing percentage of viable cells relative to PBS control in malignant cells from 3 patients with TCR Vβ2 + L-CTCL after treatment with 2 μg/mL of either anti-TCR Vβ2-PNU or hIgG1-PNU or an unconjugated anti-TCR Vβ2 antibody for 72 hours. Ordinary 1-way ANOVA was performed, and data are shown as mean ± s.e.m.; P < .001. (G) Representative flow cytometric plots showing percentage of viable malignant cells in a patient with TCR Vβ2 - CTCL after treatment with 2 μg/mL of either anti-TCR Vβ2-PNU or hIgG1-PNU or an unconjugated anti-TCR Vβ2 antibody for 72 hours. (H) Graph showing percentage of viable cells relative to PBS control in malignant cells from 2 patients with TCR Vβ2 - L-CTCL after treatment with 2 μg/mL of either anti-TCR Vβ2-PNU or hIgG1-PNU or an unconjugated anti-TCR Vβ2 antibody for 72 hours. Ordinary 1-way ANOVA was performed, and data are shown as mean ± s.e.m.; p = ns. ANOVA, analysis of variance; DAR, drug-to-antibody ratio; kDa, kilodalton; MW, molecular weight; ns, nonsignificant; s.e.m., standard error of the mean.

Article Snippet: We purchased anti-TCR Vβ2 custom antibody (REA654/MPB2D5) from Miltenyi Biotec.

Techniques: Generated, In Vitro, Conjugation Assay, Staining, SDS Page, Concentration Assay, Control, Molecular Weight

Journal: Blood Advances

Article Title: TCRVβ-targeting antibody-drug conjugates as a novel strategy to eliminate malignant T cells in T cell cancers

doi: 10.1182/bloodadvances.2026020282

Figure Lengend Snippet: TCRVβ2 is internalized by TCR Vβ2 + cells. (A) Bar plot showing the frequency of TCR Vβ2 (TRBV20-1), the most common TCRVβ (TRBV) among others in malignant T cells, in publicly available single-cell RNA sequencing data from 104 patients with CTCL. (B) Anti-TCR Vβ2-pHrodo antibody (1 μg/mL) was added to MOLT-16 (TCR Vβ2 + cells) or MyLa 2059 (TCR Vβ2 − cells). Quantification of red fluorescence over time after the addition of anti-TCR Vβ2-pHrodo antibodies to indicated cells. Data are mean ± s.e.m. of 3 technical replicates. n = 2 biological replicates. (C) MOLT-16 cells were incubated with anti-TCR Vβ2 antibody (1 μg/mL) and then analyzed using confocal microscopy. LAMP1 antibodies and DAPI (4′,6-diamidino-2-phenylindole) staining mark lysosomes and nuclei, respectively. Antihuman IgG–Alexa Fluor 488 was used to detect the location of anti-TCR Vβ2 antibodies. Scale bars, 200 μm. n = 2 biological replicates. h, hour; LAMP1, lysosomal-associated membrane protein 1; min, minutes; s.e.m, standard error of the mean.

Article Snippet: We purchased anti-TCR Vβ2 custom antibody (REA654/MPB2D5) from Miltenyi Biotec.

Techniques: Single Cell, RNA Sequencing, Fluorescence, Incubation, Confocal Microscopy, Staining, Membrane

Journal: Antibody Therapeutics

Article Title: Novel ADC and γδ T cell engager targeting CDH17 for the therapy of gastrointestinal cancers

doi: 10.1093/abt/tbag012

Figure Lengend Snippet: VHH371 binding to all subtypes of γδ TCR. (a–d). ELISA binding assays showed TAVO307 × VHH371, δ1TCR-ref, δ2TCR-ref, and γ9TCR-ref antibodies binding to recombinant human Vγ9Vδ1 (a), Vγ9Vδ2 (b), Vγ4Vδ1 (c), and Vγ4Vδ2 (d) subtypes of γδ TCR, respectively. The y -axes absorbances at 450 nm were plotted against the x -axes concentrations of the respective antibodies. Experiments were performed in duplicate with data reported as mean ± SD. (e and f) Flow cytometry-based cell binding assays showed TAVO307 × VHH371, TAVO307 × null, and null × VHH371 bispecific antibodies binding to the Vδ1 (e) and Vδ2 (f) T cells. The y -axes MFI folds over isotype control antibody were plotted against the x -axes concentrations of the test antibodies. Data shown are from a single experiment.

Article Snippet: At Day 14, the purity of the Vδ1 T cell population was assessed by flow cytometry using Alexa Fluor 488 anti-human CD3 antibody (Biolegend, 300415), PerCP-Vio 700 anti-human TCRγ/δ antibody (Miltenyi Biotec, 130-113-506), and VioBlue anti-human TCR Vδ1 antibody (Miltenyi Biotec, 130-120-583).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Recombinant, Flow Cytometry, Control

Journal: Antibody Therapeutics

Article Title: Novel ADC and γδ T cell engager targeting CDH17 for the therapy of gastrointestinal cancers

doi: 10.1093/abt/tbag012

Figure Lengend Snippet: TAVO307 × VHH371 mediated γδ T cell degranulation upon ligation to CDH17-expressing tumor cells. (a) The cartoon diagram showed γδ T cell engagers mediating γδ T cell degranulation and cytotoxicity of CDH17-expressing tumor cells. (b–e) For γδ T cell degranulation assays, testing antibodies were added to 1 × 10 5 Vδ1 or Vδ2 T cells (expanded from PBMC obtained from a single donor) co-cultured with 2 × 10 5 CDH17-expressing tumor cells (1:2 E:T ratio). After 4 h incubation, the expression of CD107a on γδ T cells was quantitated by flow cytometry. TAVO307 × VHH371, TAVO307 × null, null × VHH371 bispecific antibodies were tested for activation of CD107a degranulation marker expression in Vδ1 (b and d) or Vδ2 (c and e) T cells by SNU-5 (b and c) and AsPC-1 (d and e) cells. The y-axes percentages of CD107a-positive γδ T cells were plotted against the x -axes concentrations of the respective antibodies. Experiments were performed in duplicate or triplicate with data reported as mean ± SD.

Article Snippet: At Day 14, the purity of the Vδ1 T cell population was assessed by flow cytometry using Alexa Fluor 488 anti-human CD3 antibody (Biolegend, 300415), PerCP-Vio 700 anti-human TCRγ/δ antibody (Miltenyi Biotec, 130-113-506), and VioBlue anti-human TCR Vδ1 antibody (Miltenyi Biotec, 130-120-583).

Techniques: Ligation, Expressing, Cell Culture, Incubation, Flow Cytometry, Activation Assay, Marker

Journal: Antibody Therapeutics

Article Title: Novel ADC and γδ T cell engager targeting CDH17 for the therapy of gastrointestinal cancers

doi: 10.1093/abt/tbag012

Figure Lengend Snippet: TAVO307 × VHH371 mediated γδ T cell in the cytotoxicity of CDH17-expressing tumor cells. 2 × 10 5 γδ T cells expanded from PBMCs obtained from a single donor were co-cultured with 1 × 10 5 CDH17-expressing tumor cells (2:1 E:T ratio) in the presence of serial dilutions of testing antibodies for 24 h. The percentages of target cell lysis were determined by flow cytometry. γδ T cell cytotoxicity assays showed TAVO307 × VHH371, TAVO307 × null, null × VHH371 bispecific antibodies in mediating the cytotoxicity of SNU-5 (a and b) and AsPC-1 (c and d) cells by Vδ1 (a and c), or Vδ2 (b and d) T cells. The y -axes percentages of target cell lysis were plotted against the x -axes concentrations of the respective antibodies. Experiments were performed in duplicate with data reported as mean ± SD.

Article Snippet: At Day 14, the purity of the Vδ1 T cell population was assessed by flow cytometry using Alexa Fluor 488 anti-human CD3 antibody (Biolegend, 300415), PerCP-Vio 700 anti-human TCRγ/δ antibody (Miltenyi Biotec, 130-113-506), and VioBlue anti-human TCR Vδ1 antibody (Miltenyi Biotec, 130-120-583).

Techniques: Expressing, Cell Culture, Lysis, Flow Cytometry