Merck Exploratory Science Center, Merck & Co., Inc., Cambridge, MA, USA.
InduPro, Cambridge, MA, USA.
Nat Chem Biol. 2022 Aug;18(8):850-858. doi: 10.1038/s41589-022-01044-0. Epub 2022 Jun 2.
The growing appreciation of immune cell-cell interactions within disease environments has led to extensive efforts to develop immunotherapies. However, characterizing complex cell-cell interfaces in high resolution remains challenging. Thus, technologies leveraging therapeutic-based modalities to profile intercellular environments offer opportunities to study cell-cell interactions with molecular-level insight. We introduce photocatalytic cell tagging (PhoTag) for interrogating cell-cell interactions using single-domain antibodies (VHHs) conjugated to photoactivatable flavin-based cofactors. Following irradiation with visible light, the flavin photocatalyst generates phenoxy radical tags for targeted labeling. Using this technology, we demonstrate selective synaptic labeling across the PD-1/PD-L1 axis in antigen-presenting cell-T cell systems. In combination with multiomics single-cell sequencing, we monitored interactions between peripheral blood mononuclear cells and Raji PD-L1 B cells, revealing differences in transient interactions with specific T cell subtypes. The utility of PhoTag in capturing cell-cell interactions will enable detailed profiling of intercellular communication across different biological systems.
免疫细胞-细胞相互作用在疾病环境中的作用日益受到重视，这促使人们大力开发免疫疗法。然而，以高分辨率描绘复杂的细胞-细胞界面仍然具有挑战性。因此，利用基于治疗的模式来描绘细胞间环境的技术为研究细胞-细胞相互作用提供了机会，可以深入了解分子水平上的相互作用。我们引入光催化细胞标记（PhoTag）技术，使用与光活化黄素辅因子偶联的单域抗体（VHH）来研究细胞-细胞相互作用。可见光照射后，黄素光催化剂生成苯氧自由基标签，用于靶向标记。利用这项技术，我们在抗原呈递细胞- T 细胞系统中对 PD-1/PD-L1 轴上的突触进行了选择性标记。结合多组学单细胞测序，我们监测了外周血单核细胞与 Raji PD-L1 B 细胞之间的相互作用，揭示了与特定 T 细胞亚型的短暂相互作用的差异。PhoTag 在捕获细胞-细胞相互作用方面的应用将能够详细描绘不同生物系统中的细胞间通讯。
