Department of Genetics, Stanford University, Stanford, CA, USA.
Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA, USA.
Nat Methods. 2022 Oct;19(10):1295-1305. doi: 10.1038/s41592-022-01592-2. Epub 2022 Sep 5.
Adaptive immunity relies on T lymphocytes that use αβ T cell receptors (TCRs) to discriminate among peptides presented by major histocompatibility complex molecules (pMHCs). Identifying pMHCs capable of inducing robust T cell responses will not only enable a deeper understanding of the mechanisms governing immune responses but could also have broad applications in diagnosis and treatment. T cell recognition of sparse antigenic pMHCs in vivo relies on biomechanical forces. However, in vitro screening methods test potential pMHCs without force and often at high (nonphysiological) pMHC densities and thus fail to predict potent agonists in vivo. Here, we present a technology termed BATTLES (biomechanically assisted T cell triggering for large-scale exogenous-pMHC screening) that uses biomechanical force to initiate T cell triggering for peptides and cells in parallel. BATTLES displays candidate pMHCs on spectrally encoded beads composed of a thermo-responsive polymer capable of applying shear loads to T cells, facilitating exploration of the force- and sequence-dependent landscape of T cell responses. BATTLES can be used to explore basic T cell mechanobiology and T cell-based immunotherapies.
适应性免疫依赖于 T 淋巴细胞，其利用 αβ T 细胞受体 (TCR) 来区分主要组织相容性复合物分子 (pMHC) 呈递的肽。鉴定能够诱导强烈 T 细胞反应的 pMHC，不仅将使我们能够更深入地了解免疫反应的调控机制，而且还可能在诊断和治疗方面具有广泛的应用。T 细胞在体内识别稀疏的抗原性 pMHC 依赖于生物力学力。然而，体外筛选方法在没有力的情况下测试潜在的 pMHC，并且通常在高 (非生理) pMHC 密度下进行，因此无法预测体内的有效激动剂。在这里，我们提出了一种称为 BATTLES（大规模外源性 pMHC 筛选的生物力学辅助 T 细胞触发）的技术，该技术利用生物力学力平行启动肽和细胞的 T 细胞触发。BATTLES 将候选 pMHC 展示在由热响应聚合物组成的光谱编码珠上，该聚合物能够对 T 细胞施加剪切力，从而促进探索 T 细胞反应的力依赖性和序列依赖性景观。BATTLES 可用于探索基本的 T 细胞机械生物学和基于 T 细胞的免疫疗法。
