Division of Rheumatology, Department of Internal Medicine III, Medical University of Vienna, Vienna, Austria.
Front Immunol. 2023 Mar 24;14:1096096. doi: 10.3389/fimmu.2023.1096096. eCollection 2023.
Various autoimmune diseases are characterized by distinct cell subset distributions and activation profiles of peripheral blood mononuclear cells (PBMCs). PBMCs can therefore serve as an ideal biomarker material, which is easily accessible and allows for screening of multiple cell types. A detailed understanding of the immune landscape is critical for the diagnosis of patients with autoimmune diseases, as well as for a personalized treatment approach. In our study, we investigate the potential of multi-parameter spectral flow cytometry for the identification of patients suffering from autoimmune diseases and its power as an evaluation tool for drug screening approaches (advanced immunophenotyping). We designed a combination of two 22-color immunophenotyping panels for profiling cell subset distribution and cell activation. Downstream bioinformatics analyses included percentages of individual cell populations and median fluorescent intensity of defined markers which were then visualized as heatmaps and in dimensionality reduction approaches. testing of epigenetic immunomodulatory drugs revealed an altered activation status upon treatment, which supports the use of spectral flow cytometry as a high-throughput drug screening tool. Advanced immunophenotyping might support the exploration of novel therapeutic drugs and contribute to future personalized treatment approaches in autoimmune diseases and beyond.
各种自身免疫性疾病的特征是外周血单个核细胞（PBMC）中不同的细胞亚群分布和激活谱。因此，PBMC 可以作为一种理想的生物标志物材料，易于获取，并允许对多种细胞类型进行筛选。详细了解免疫景观对于自身免疫性疾病患者的诊断以及个性化治疗方法至关重要。在我们的研究中，我们研究了多参数光谱流式细胞术在识别患有自身免疫性疾病的患者中的潜力及其作为药物筛选方法（高级免疫表型分析）的评估工具的能力。我们设计了两个 22 色免疫表型分析面板的组合，用于分析细胞亚群分布和细胞激活。下游的生物信息学分析包括各个细胞群体的百分比和定义标记物的中荧光强度，然后以热图和降维方法可视化。对表观遗传免疫调节药物的测试显示，治疗后激活状态发生改变，这支持将光谱流式细胞术用作高通量药物筛选工具。高级免疫表型分析可能有助于探索新的治疗药物，并为自身免疫性疾病及其他疾病的未来个性化治疗方法做出贡献。
