Department of Rheumatology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands.
Department of Synthetic Organic Chemistry, Radboud University, Heyendaalseweg 135, 6525 AJ Nijmegen, The Netherlands.
ACS Biomater Sci Eng. 2022 Apr 11;8(4):1486-1493. doi: 10.1021/acsbiomaterials.1c01395. Epub 2022 Mar 8.
Multivalent scaffolds that carry multiple molecules with immunophenotyping or immunomodulatory properties are invaluable tools for studying and modulating specific functions of human immune responses. So far, streptavidin-biotin-based tetramers have been widely used for B-cell immunophenotyping purposes. However, the utility of these tetramers is limited by their tetravalency, the inherent immunogenicity of streptavidin (a bacterial protein that can potentially be recognized by B cells), and the limited feasibility to functionalize these reagents. This has rendered tetramers suboptimal for studying rare, in particular, antigen-specific B-cell populations in the context of clinical applications. Here, we used polyisocyanopeptides (PICs), multivalent polymeric scaffolds functionalized with around 50 peptide antigens, to detect autoreactive B cells in the peripheral blood of patients with rheumatoid arthritis. To explore the potential immunomodulatory functionalities, we functionalized PICs with autoantigenic peptides and a trisaccharide CD22 ligand to inhibit autoreactive B-cell activation through interference with the B-cell receptor activation pathway, as evidenced by reduced phospho-Syk expression upon PIC binding. Given the possibilities to functionalize PICs, our data demonstrate that the modular and versatile character of PIC scaffolds makes them promising candidates for future clinical applications in B-cell-mediated diseases.
多价支架携带具有免疫表型或免疫调节特性的多种分子,是研究和调节人类免疫反应特定功能的宝贵工具。到目前为止,基于链霉亲和素-生物素的四聚体已被广泛用于 B 细胞免疫表型分析目的。然而,这些四聚体的用途受到其四价性、链霉亲和素(一种潜在可被 B 细胞识别的细菌蛋白)的固有免疫原性以及对这些试剂进行功能化的有限可行性的限制。这使得四聚体在研究罕见的、特别是在临床应用背景下的抗原特异性 B 细胞群体方面不太理想。在这里,我们使用多异氰酸酯肽(PICs),即用约 50 个肽抗原功能化的多价聚合物支架,来检测类风湿关节炎患者外周血中的自身反应性 B 细胞。为了探索潜在的免疫调节功能,我们用自身抗原肽和三糖 CD22 配体对 PICs 进行功能化,通过干扰 B 细胞受体激活途径来抑制自身反应性 B 细胞的激活,这表现在 PIC 结合后磷酸化 Syk 的表达减少。鉴于 PIC 可进行功能化的可能性,我们的数据表明,PIC 支架的模块化和多功能特性使其成为未来在 B 细胞介导的疾病中进行临床应用的有希望的候选者。