Murai Aiko, Iwata Masashi, Miyakawa Shuuichi, Warude Dnyaneshwar, Sagara Masaki, Kikukawa Yusuke
Research, Oncology Drug Discovery Unit, Takeda Pharmaceutical Company Limited, Fujisawa, Kanagawa, 251-8555, Japan.
Research, Oncology Drug Discovery Unit, Takeda Pharmaceutical Company Limited, Fujisawa, Kanagawa, 251-8555, Japan.
Cytotherapy. 2025 May;27(5):571-580. doi: 10.1016/j.jcyt.2024.12.015. Epub 2025 Jan 6.
The dysregulation of follicular helper T (Tfh) cell function, followed by the proliferation of self-reactive B cells, can lead to the development of autoimmune diseases. Recently, adaptive T regulatory cell (Treg) transfer therapy has attracted considerable attention for inducing effective immune tolerance owing to Tregs' diverse immune-inhibitory activities. However, preclinical studies and recent clinical trials of polyclonal Treg therapy have suggested further improving the efficacy of Treg therapy through targeted tissue specificity and local persistence by gene engineering. In this study, we reported a novel approach to specifically inhibit Tfh cells by CXC motif chemokine receptor 5-targeted chimeric antigen receptor (CXCR5-CAR) Tregs.
Tregs expressing CAR against CXCR5 were generated from human peripheral blood mononuclear cells-derived Tregs. The phenotype and suppressive capacity of the engineered Tregs were evaluated using coculture assays with naïve T cells, circulating Tfh (cTfh) cells, or a combination of cTfh cells and naïve B cells through flow cytometry analysis.
CXCR5-CAR Tregs induced more potent inhibition of circulating cTfh cell proliferation while maintaining similar suppressive properties on CXCR5-negative responder cells compared with non-selective polyclonal Tregs. The antigen-dependent activation of CXCR5-CAR Tregs was confirmed by latency-associated peptide (LAP) expression in the coculture with cTfh cells. In the coculture condition with both cTfh and naïve B cells, the activation of naïve B cells induced by cTfh cells was more effectively inhibited by CXCR5-CAR Tregs than by polyclonal Tregs.
The results demonstrate the potential of CXCR5-CAR Tregs to effectively inhibit the Tfh-B cell response in autoimmune diseases, paving the way for further research to confirm their functional superiority in vivo. This novel approach offers promise for achieving local, long-term immune tolerance compared with existing approaches such as nonspecific immunosuppression and polyclonal Treg therapy.
滤泡辅助性T（Tfh）细胞功能失调，随后自身反应性B细胞增殖，可导致自身免疫性疾病的发生。近来，适应性调节性T细胞（Treg）转移疗法因其多样的免疫抑制活性，在诱导有效免疫耐受方面备受关注。然而，多克隆Treg疗法的临床前研究和近期临床试验表明，可通过基因工程实现靶向组织特异性和局部持久性，进一步提高Treg疗法的疗效。在本研究中，我们报告了一种通过靶向CXC基序趋化因子受体5的嵌合抗原受体（CXCR5-CAR）Treg特异性抑制Tfh细胞的新方法。
从人外周血单个核细胞来源的Treg中产生表达抗CXCR5的CAR的Treg。通过流式细胞术分析，利用与初始T细胞、循环Tfh（cTfh）细胞或cTfh细胞与初始B细胞组合的共培养试验，评估工程化Treg的表型和抑制能力。
与非选择性多克隆Treg相比，CXCR5-CAR Treg在抑制循环cTfh细胞增殖方面更有效，同时对CXCR5阴性应答细胞保持相似的抑制特性。在与cTfh细胞共培养中，潜伏相关肽（LAP）表达证实了CXCR5-CAR Treg的抗原依赖性激活。在cTfh细胞和初始B细胞共培养条件下，与多克隆Treg相比，CXCR5-CAR Treg能更有效地抑制cTfh细胞诱导的初始B细胞激活。
结果表明CXCR5-CAR Treg在自身免疫性疾病中有效抑制Tfh-B细胞反应的潜力，为进一步研究证实其体内功能优势铺平了道路。与非特异性免疫抑制和多克隆Treg疗法等现有方法相比，这种新方法有望实现局部、长期的免疫耐受。
