Siebrand Cynthia J, Bergo Nicholas J, Lee Suckwon, Andersen Julie K, Walton Chaska C
Buck Institute for Research On Aging, 8001 Redwood Blvd., Novato, CA, 94945, USA.
USC Leonard Davis School of Gerontology, University of Southern California, 3715 McClintock Ave, Los Angeles, CA, 90191, USA.
J Transl Med. 2025 May 30;23(1):605. doi: 10.1186/s12967-025-06572-6.
The lack of a definitive cure for Alzheimer's disease (AD) is fueling the search for innovative therapeutic strategies. Having revolutionized cancer immunotherapy, immune cell engineering with chimeric antigen receptors (CAR) is being explored to target AD. Whether CARs can recognize distinct amyloid-β (Aβ) species and tau neurofibrillary tangles (NFTs)-hallmark pathologies of AD-remains unclear.
To investigate this, we engineered a series of CARs using single-chain fragment variable (scFv) derived from the variable light and heavy chains of antibodies tested in AD clinical trials. These included E2814 (E2814-CAR), targeting tau; Lecanemab (Lec-CAR) and Aducanumab (Adu-CAR), targeting Aβ; and Donanemab (Don-CAR) and Remternetug (Rem-CAR), targeting the truncated pyroglutamated Aβ species Aβp3-42. To evaluate CAR function, we utilized the murine DO11.10 CD4⁺ T-cell hybridoma line as a scalable and reproducible platform. CAR activation was assessed in response to tau preformed fibrils (PFFs), Aβ oligomer-enriched aggregates, and Aβp3-42 aggregates, using flow cytometry for CD69 expression and ELISA for IL-2 secretion. To validate this platform, we tested Adu-CAR in primary mouse CD4⁺ T cells treated with Aβ aggregates and assessed activation via flow cytometry for CD69 and CD25 expression.
DO11.10 cells expressing E2814-CAR-but not Lec-CAR-responded to tau PFFs. In contrast, cells expressing Adu-CAR, and to a lesser extent Lec-CAR-but not E2814-CAR-responded to Aβ aggregates. For Aβp3-42 aggregates, Rem-CAR elicited the strongest response, followed by Adu-CAR, while E2814-CAR and Don-CAR showed no activation. The activation of Adu-CAR by Aβ aggregates was recapitulated in primary CD4⁺ T cells, as measured by CD69 expression.
Our findings demonstrate that CARs can detect and discriminate between tau PFFs, Aβ1-42, and Aβp3-42 aggregates. This highlights the potential of repurposing AD antibodies for CAR-based therapies to selectively target tau NFTs and distinct forms of Aβ senile plaques.
阿尔茨海默病（AD）缺乏明确的治愈方法，这推动了对创新治疗策略的探索。嵌合抗原受体（CAR）免疫细胞工程技术彻底改变了癌症免疫疗法，目前正在探索将其用于治疗AD。CAR是否能够识别AD的标志性病理特征——不同种类的淀粉样β蛋白（Aβ）和tau神经原纤维缠结（NFT）——仍不清楚。
为了研究这一问题，我们使用在AD临床试验中测试过的抗体可变轻链和重链衍生的单链可变片段（scFv）构建了一系列CAR。这些包括靶向tau的E2814（E2814-CAR）；靶向Aβ的lecanemab（Lec-CAR）和aducanumab（Adu-CAR）；以及靶向截短的焦谷氨酸化Aβ物种Aβp3-42的多奈单抗（Don-CAR）和瑞纳替尼（Rem-CAR）。为了评估CAR的功能，我们利用小鼠DO11.10 CD4⁺ T细胞杂交瘤系作为一个可扩展且可重复的平台。使用流式细胞术检测CD69表达和ELISA检测IL-2分泌，评估CAR对tau预形成纤维（PFF）、富含Aβ寡聚体的聚集体和Aβp3-42聚集体的激活情况。为了验证这个平台，我们在用Aβ聚集体处理的原代小鼠CD4⁺ T细胞中测试了Adu-CAR，并通过流式细胞术检测CD69和CD25表达来评估激活情况。
表达E2814-CAR的DO11.10细胞——而不是Lec-CAR——对tau PFF有反应。相比之下，表达Adu-CAR的细胞，以及程度较轻的表达Lec-CAR的细胞——而不是E2814-CAR——对Aβ聚集体有反应。对于Aβp3-42聚集体，Rem-CAR引发的反应最强，其次是Adu-CAR，而E2814-CAR和Don-CAR没有显示出激活。通过CD69表达测量，在原代CD4⁺ T细胞中重现了Aβ聚集体对Adu-CAR的激活。
我们的研究结果表明，CAR可以检测并区分tau PFF、Aβ1-42和Aβp3-42聚集体。这突出了将AD抗体重新用于基于CAR的疗法以选择性靶向tau NFT和不同形式的Aβ老年斑的潜力。
