Chimeric Antigen Receptor (CAR) T Cells and Bispecific T Cell Engagers (BiTEs).

Overview

Immunology, Medical College of Wisconsin Chimeric Antigen Receptor (CAR) T Cells and Bispecific T Cell Engagers (BiTEs) have revolutionized the treatment of relapsed/refractory hematologic cancers. However, there are no head-to-head comparisons testing differences in their ability to induce Tcel l activation - a surrogate for their efficacy and toxicity. We hypothesized that differences in downstream signaling by anti -CD19 CAR -Tcells and anti -CD3/CD19 BiTEs will lead to differential Tcell functional capacity and transcriptional profiles. Spectral flow cytometry identified the optimal activation bead, and the Beacon Optofluidic system was used to analyze differences in phenotype, functional capacity, and transcriptional profiles between CAR Tcells and Tcells co -cultured with BiTEs. Signal seekin g and validation experiments were performed to develop an analytic pipeline. Functional analysis demonstrated differential expression of cytokines (IFN -γ, TNF-𝛼, Granzyme B, Perforin) in CAR Tcells compared to Tcells with BiTEs . CAR Tcells showed greater polyfunctionality in cytokine secretion, more durable target engagement, increased likelihood of exhaustion, and higher CD8+ to CD4+ Tcell ratios than Tcells with BiTEs. Single cell RNA sequencing demonstrated a transcriptional basis for the differential cytokine expression between CAR Tcells and Tcells with BiTEs, which was confirmed by concordant protein expression at single cell resolution. The distinct mechanistic differences between CAR Tcells and BiTEs indicate the n eed to balance toxicity with therapeutic efficacy in treatment selection. We demonstrate the feasibility of developing an analytic pipeline to analyze functional, transcriptional, and proteomic data at single cell resolution to help personalize cellular therapies.

Competition history

• JSHS 2025 Category not listed

Resources

• National JSHS abstract book ↗

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