Immune function assessing of TIM3/CD28-modified CD19 CAR-T cells and general CD19 CAR-T cells through a highthroughput single-cell microarray platform

Abstract

Chimeric antigen receptor (CAR) T cells are widely used to treat hematological tumors due to their powerful ability to target and kill cancer cells, of which accurate function evaluation at the single-cell level is crucial to ensuring the efficacy of diagnosis and treatment. Currently, a universal platform to evaluate the efficacy of immune single cells rapidly, efficiently, and systematically is urgently needed. Here, the cytotoxicity, proliferative potential, and persistence of TIM3/CD28-modified CD19 CAR-T cells are evaluated in comparison with ordinary CD19 CAR-T cells through high-performance and throughput graphene oxide quantum dot (GOQD)-based single-cell microfluidic chips. Overall secretory factor expression, immune-therapy effect of different effector-target ratios, spatial immune-therapy effects, and subgroup type profiling are demonstrated to explicit the immunotherapy effect of TIM3/CD28-modified CD19 CAR-T cells. TIM3/CD28-modified CD19 CAR-T cells show stronger anti-tumor ability and maintain excellent immunotherapy effects even at low effector-target ratios and remote distances. TIM3/CD28 also strengthens the local targeting ability of TIM3/CD28-modified CD19 CAR-T. Importantly, TIM3/CD28-modified CD19 CAR-T exhibits more distinct Th1/Th2 long-term persistent and potent killer subgroups, which is very helpful for personalized therapy. Overall, this study provides a valuable approach that can be widely implemented to analyze current CAR-T combinations and evaluate the function of innovative CAR treatments in the future.

Keywords

CAR-T / immunotherapy effect evaluation / microfluidic chips / secreted factors / single cell

RIGHTS & PERMISSIONS

2024 2024 The Authors. Interdisciplinary Medicine published by Wiley-VCH GmbH on behalf of Nanfang Hospital, Southern Medical University.