德国维尔茨堡大学Markus Sauer等研究人员合作
研究人员表示，阐明膜蛋白与抗体之间的相互作用需要在高时空分辨率下进行全细胞成像。光格光片显微术（LLS）提供了快速的体积成像，但其空间分辨率有限。基于DNA的成像点累积纳米拓扑学（DNA-PAINT）可实现分子级分辨率，但由于长时间的获取过程，它仅限于二维成像。
研究人员开发了双染料成像探针（TDI），使得成像速度提高约15倍。将TDI-DNA-PAINT和LLS显微术结合应用于免疫学B细胞，研究人员揭示了内源性CD20与治疗性单克隆抗体（mAb）利妥昔单抗、奥法木单抗和奥比木单抗的寡聚状态和相互作用。
结果表明，CD20在微绒毛上大量表达，并与单克隆抗体结合，这导致抗体浓度依赖性的B细胞极化和微绒毛突起的稳定。这些发现有助于合理设计改进的免疫疗法，以靶向肿瘤相关抗原。
附：英文原文
Title: Decoding the molecular interplay of CD20 and therapeutic antibodies with fast volumetric nanoscopy
Author: Arindam Ghosh, Mara Meub, Dominic A. Helmerich, Julia Weingart, Patrick Eiring, Thomas Nerreter, K. Martin Kortüm, Sren Doose, Markus Sauer
Issue&Volume: 2025-01-10
Abstract: Elucidating the interaction between membrane proteins and antibodies requires whole-cell imaging at high spatiotemporal resolution. Lattice light-sheet (LLS) microscopy offers fast volumetric imaging but suffers from limited spatial resolution. DNA-based point accumulation for imaging in nanoscale topography (DNA-PAINT) achieves molecular resolution but is restricted to two-dimensional imaging owing to long acquisition times. We have developed two-dye imager (TDI) probes that enable ~15-fold faster imaging. Combining TDI-DNA-PAINT and LLS microscopy on immunological B cells revealed the oligomeric states and interaction of endogenous CD20 with the therapeutic monoclonal antibodies (mAbs) rituximab, ofatumumab, and obinutuzumab. Our results demonstrate that CD20 is abundantly expressed on microvilli that bind mAbs, which leads to an antibody concentration–dependent B cell polarization and stabilization of microvilli protrusions. These findings could aid rational design of improved immunotherapies targeting tumor-associated antigens.
DOI: adq4510
Source: https://www.science.org/doi/10.1126/science.adq4510