AUTHOR=Zheng Rui , Chen Yuankun , Zhang Yiting , Liang Sixin , Zhao Xiaojuan , Wang Yiyi , Wang Pengju , Meng Ruotong , Yang Angang , Yan Bo TITLE=Humanized single-domain antibody targeting HER2 enhances function of chimeric antigen receptor T cells JOURNAL=Frontiers in Immunology VOLUME=14 YEAR=2023 URL=https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2023.1258156 DOI=10.3389/fimmu.2023.1258156 ISSN=1664-3224 ABSTRACT=Introduction

Chimeric antigen receptors (CARs) can redirect T cells against antigen-expressing tumors, and each component plays an important role in the function and anti-tumor efficacy. It has been reported that using human sequences or a low affinity of CAR single-chain variable fragments (scFvs) in the CAR binding domains is a potential way to enhance the function of CAR-T cells. However, it remains largely unknown how a lower affinity of CARs using humanized scFvs affects the function of CAR-T cells until recently.

Methods

We used different humanized anti-HER2 antibodies as the extracellular domain of CARs and further constructed a series of the CAR-T cells with different affinity.

Results

We have observed that moderately reducing the affinity of CARs (light chain variable domain (VL)-based CAR-T) could maintain the anti-tumor efficacy, and improved the safety of CAR therapy both in vitro and in vivo compared with high-affinity CAR-T cells. Moreover, T cells expressing the VL domain only antibody exhibited long-lasting tumor elimination capability after multiple challenges in vitro, longer persistence and lower cytokine levels in vivo.

Discussion

Our findings provide an alternative option for CAR-T optimization with the potential to widen the use of CAR T cells.