Journal: Frontiers in Immunology

Article Title: Logic-gated and contextual control of immunotherapy for solid tumors: contrasting multi-specific T cell engagers and CAR-T cell therapies

doi: 10.3389/fimmu.2024.1490911

Figure Lengend Snippet: Architectures for OR and AND-gate bispecific CARs. (A) OR gates can respond to both TAAs either singly or together, offering an opportunity to restrict antigen escape. Two variations are shown; dual CARs which can be encoded from the same bicistronic vector, or delivered by two separate vectors, allow the use of complementary costimulation domains as shown here. Tandem CARs contain binding domains targeting two distinct antigens within the same CAR. (B) A true AND gate is only activated when both targets are present, and do not respond to cells expressing only one target. The split CAR concept places the domains that drive signal 1 (activation via the CD3z ITAM) and signal 2 (costimulation, shown here with the CD28 ITAM) on separate CARs. A more recent, and potentially more tightly controlled format are the LINK CARs, which place two effectors that lie downstream of the TCR (LAT and SLP76), on two distinct CARs, thus splitting signal 1 in two and requiring engagement of both CARs to initiate T-cell activation. Created with BioRender.com .

Article Snippet: A company called BioAtla has recently leveraged 4- to 5-fold affinity improvement at low pH for EPCAM and CD3 binders (engineered through addition of histidine switches at the CDR/antigen interface) to generate a dual Conditionally Activated Bispecific (CAB) TCE against EPCAM ( ) with a 10-fold in vitro killing window between pH 6.5 versus 7.4 ( ).

Techniques: Plasmid Preparation, Binding Assay, Expressing, Activation Assay

Journal: Cancers

Article Title: Unlocking Intracellular Oncology Targets: The Unique Role of Antibody-Based T-Cell Receptor Mimic (TCRm) Therapeutics in T-Cell Engagers (TCEs) and Antibody-Drug Conjugates (ADCs)

doi: 10.3390/cancers16223776

Figure Lengend Snippet: List of recent TCRm therapeutics in preclinical and clinical development.

Article Snippet: HIV/A2 , TCE bispecific , Preclinical , Johns Hopkins University , [ ] .

Techniques:

Journal: Cancers

Article Title: Unlocking Intracellular Oncology Targets: The Unique Role of Antibody-Based T-Cell Receptor Mimic (TCRm) Therapeutics in T-Cell Engagers (TCEs) and Antibody-Drug Conjugates (ADCs)

doi: 10.3390/cancers16223776

Figure Lengend Snippet: List of recent TCRm therapeutics in preclinical and clinical development.

Article Snippet: MAGE-A4/A2 , TCE bispecific , Phase 1 , Genentech , Clinicaltrials.gov (NCT06372574).

Techniques:

Journal: bioRxiv

Article Title: Dual Targeting for Enhanced Tumor Immunity: Conditionally Active CD28xVISTA Bispecific Antibodies Promote Myeloid-Driven T-Cell Activation

doi: 10.1101/2025.04.07.647657

Figure Lengend Snippet: CD28xVISTA bsAb induces IL-2–luciferase reporter gene expression both in cis and in trans . A, Luciferase expression from a Jurkat IL-2-luciferase reporter cell line as a function of concentration of CD28xVISTA BS1 (green) or parental monospecific mAbs anti-VISTA (orange) or anti-CD28 (blue). B, Luciferase expression from the reporter cell line co-cultured with a HEK293 cell line expressing anti-CD3 (OKT3-scFv) alone (open symbols; left in schematic) or in combination with VISTA (filled symbols; right in schematic) enabling co-stimulation by a CD28xVISTA bsAb in cis . C, Luciferase expression from the IL-2 reporter cell line as a function of CD28xVISTA BS1 when co-cultured with the HEK293 cell line expressing anti-CD3 (OKT3-scFv) in combination with a CHO cell line overexpressing VISTA (orange) enabling co-stimulation by a CD28xVISTA bsAb in trans . A parallel negative control with the parental CHO cells substituting for CHO-VISTA cells (blue) or a positive control with the HEK293 cell line co-expressing anti-CD3 (OKT3-scFv) and VISTA (allowing for cis co-stimulation; green) are indicated.

Article Snippet: Human T-cell mediated killing of LNCaP prostate cancer cells was analyzed on the xCELLigence real-time cell analysis platform (Agilent) by co-culturing LNCaP cells with PBMCs and VISTA + Kasumi-3 cells in the presence of a CD3xPSMA bispecific T-cell engager (BPS Bioscience; Cat. # 101242-2 with a CD3-binding domain from the CD3xCD20 TCE Mosunetuzumab) alone or in combination with CD28xVISTA BS2.

Techniques: Luciferase, Gene Expression, Expressing, Concentration Assay, Cell Culture, Negative Control, Positive Control

Journal: bioRxiv

Article Title: Dual Targeting for Enhanced Tumor Immunity: Conditionally Active CD28xVISTA Bispecific Antibodies Promote Myeloid-Driven T-Cell Activation

doi: 10.1101/2025.04.07.647657

Figure Lengend Snippet: In vitro human T-cell co-stimulation in trans by CD28xVISTA BS2 (non-pH sensitive) potentiates LNCaP killing by a CD3xPSMA T-cell engager. A, Schematic representation of LNCaP prostate cancer cells co-cultured with Kasumi-3 and T-cells from human PBMCs. LNCaP killing by a CD3xPSMA TCE can potentially be enhanced through CD28 trans co-stimulation by CD28xVISTA BS2 B, Signal resulting from LNCaP cells in the Agilent xCELLigence assay platform in the absence of mAbs (green squares), in the presence of increasing sub-optimal concentrations of CD3xPSMA alone (4 ng/ml (orange), 12 ng/ml (light green) or 37 ng/ml (blue)); or same concentrations of CD3xPSMA in the presence of 0.1 μg/ml CD28xVISTA BS2 (purple, black, and blue triangles, respectively). A control with lysis buffer added to co-culture wells is indicated by gray. Only growth of the adherent LNCaP cells caused a change in the impedance signal measured by the xCELLigence platform. C, IL-2 or TNF-α cytokine release measured Day 1, ( D ) T-cell activation measured as %CD25 + of total T-cells on Day 6 and ( E ) T-cell proliferation enumerated as T-cell count per volume on Day 6 in the absence (blue) or presence of 0.1 μg/ml CD28xVISTA BS2 (orange) (mean values +/-SD; n=3 technical replicates).

Article Snippet: Human T-cell mediated killing of LNCaP prostate cancer cells was analyzed on the xCELLigence real-time cell analysis platform (Agilent) by co-culturing LNCaP cells with PBMCs and VISTA + Kasumi-3 cells in the presence of a CD3xPSMA bispecific T-cell engager (BPS Bioscience; Cat. # 101242-2 with a CD3-binding domain from the CD3xCD20 TCE Mosunetuzumab) alone or in combination with CD28xVISTA BS2.

Techniques: In Vitro, Cell Culture, Control, Lysis, Co-Culture Assay, Activation Assay, Cell Counting