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高通量磁珠法细胞因子谱揭示 CAR-T 治疗中的动态免疫特征并鉴定候选探索性生物标志物

英文原题:High-throughput bead-based cytokine profiling reveals dynamic immune signatures and identifies candidate exploratory biomarkers in CAR-T therapy.

PubMed 2026/07/29(内容时间) Chem Sci Q1 · IF 8.1(JCR 2025)

研究概要

这些发现证明了可扩展的基于微珠的平台在CAR-T治疗中进行高分辨率免疫监测的实用性,并确定了基线TGF-β1作为与CRS严重程度和临床结局相关的候选探索性生物标志物。

中文摘要

CAR-T 细胞疗法已经彻底改变了难治性B细胞恶性肿瘤的治疗,但严重细胞因子释放综合征(CRS)和不可预测的长期结局仍是主要临床挑战。在此,我们开发了一种快速、可扩展、基于96孔板的策略,用于高通量制备镧系元素编码的分类器微珠。该方法将生产通量从每天几种大幅提高到8小时内96种,使得通过质谱流式细胞术进行灵敏的多重细胞因子谱分析成为可能。通过将该平台应用于32例接受CD19靶向CAR-T治疗的弥漫性大B细胞淋巴瘤患者的136份纵向血浆样本,我们在14种关键细胞因子中实现了fg mL -1水平的灵敏度和宽动态范围。纵向分析揭示了低级别和高级别CRS患者之间不同的细胞因子动态变化。基线TGF- 1显示与后续高级别CRS(p = 0.001)和较差的无事件生存期(p = 0.02)存在显著的单变量关联。这些发现证明了可扩展的基于微珠的平台在CAR-T治疗中用于高分辨率免疫监测的实用性,并将基线TGF- 1确定为与CRS严重程度和临床结局相关的候选探索性生物标志物。该方法为灵敏且高通量的细胞因子谱分析以及更深入理解免疫治疗中的免疫反应提供了一种实用工具。

展开英文摘要原文

Chimeric antigen receptor T-cell (CAR-T) therapy has revolutionized the treatment of refractory B-cell malignancies, yet severe cytokine release syndrome (CRS) and unpredictable long-term outcomes remain major clinical challenges. Here, we developed a rapid, scalable, 96-well plate-based strategy for the high-throughput preparation of lanthanide-encoded classifier beads. This approach substantially increases production throughput, from several types per day to 96 types within 8 hours, enabling sensitive multiplexed cytokine profiling by mass cytometry. By applying this platform to 136 longitudinal plasma samples from 32 patients with diffuse large B-cell lymphoma receiving CD19-directed CAR-T therapy, we achieved fg mL -1 -level sensitivity and broad dynamic ranges across 14 key cytokines. Longitudinal profiling revealed distinct cytokine dynamics between patients with low- and high-grade CRS. Baseline TGF- 1 showed a significant univariate association with subsequent high-grade CRS ( p = 0.001) and inferior event-free survival ( p = 0.02). These findings demonstrate the utility of a scalable bead-based platform for high-resolution immune monitoring in CAR-T therapy and identify baseline TGF- 1 as a candidate exploratory biomarker associated with CRS severity and clinical outcome. This approach provides a practical tool for sensitive and high-throughput cytokine profiling and deeper understanding of immune responses in immunotherapy.

论文信息

作者
Pu K、He W、Feng J、Li H、Chan CL、Chan TSY、Tse E、Sun H
单位
Department of Chemistry and HKU-CAS Joint Laboratory of Metallomics on Health and Environment, The University of Hong Kong Pokfulam Road Hong Kong SAR China hsun@hku.hk.Hong Kong
期刊
Chemical science2026 Sep 2
原文标识
PubMed 42529470 · DOI 10.1039/d6sc03656b