Koo Doyeon, Mao Zhiyuan, Dimatteo Robert, Tsubamoto Natalie, Noguchi Miyako, McLaughlin Jami, Tran Wendy, Lee Sohyung, Cheng Donghui, de Rutte Joseph, Sojo Giselle Burton, Witte Owen N, Di Carlo Dino
Department of Bioengineering, University of California, Los Angeles; Los Angeles, CA 90095, USA.
Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles; Los Angeles, CA 90095, USA.
bioRxiv. 2023 Jan 20:2023.01.17.524440. doi: 10.1101/2023.01.17.524440.
The ability to selectively bind to antigenic peptides and secrete cytokines can define populations of cells with therapeutic potential in emerging T cell receptor (TCR) immunotherapies. We leverage cavity-containing hydrogel microparticles, called nanovials, each coated with millions of peptide-major histocompatibility complex (pMHC) monomers to isolate antigen-reactive T cells. T cells are captured and activated by pMHCs and secrete cytokines on nanovials, allowing sorting based on both affinity and function. The TCRs of sorted cells on nanovials are sequenced, recovering paired αβ-chains using microfluidic emulsion-based single-cell sequencing. By labeling nanovials having different pMHCs with unique oligonucleotide-barcodes we could link TCR sequence to targets with 100% accuracy. We identified with high specificity an expanded repertoire of functional TCRs targeting viral antigens compared to standard techniques.
在新兴的T细胞受体（TCR）免疫疗法中，能够选择性结合抗原肽并分泌细胞因子可定义具有治疗潜力的细胞群体。我们利用含有腔室的水凝胶微粒（称为纳米小瓶），每个纳米小瓶都包被有数百万个肽 - 主要组织相容性复合体（pMHC）单体，以分离抗原反应性T细胞。T细胞被pMHC捕获并激活，并在纳米小瓶上分泌细胞因子，从而能够基于亲和力和功能进行分选。对纳米小瓶上分选细胞的TCR进行测序，使用基于微流控乳液的单细胞测序回收配对的αβ链。通过用独特的寡核苷酸条形码标记具有不同pMHC的纳米小瓶，我们可以100%准确地将TCR序列与靶标联系起来。与标准技术相比，我们以高特异性鉴定出了靶向病毒抗原的功能性TCR的扩展库。
