Department of Hematology, Oncology, and Tumor Immunology, Charite´ - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany.
Berlin Institute of Health, Charité - Universitätsmedizin Berlin, BIH Center for Regenerative Therapies (BCRT), Berlin, Germany.
Front Immunol. 2022 May 23;13:874499. doi: 10.3389/fimmu.2022.874499. eCollection 2022.
Success and complications of allogeneic hematopoietic stem cell transplantation (alloHSCT) are closely connected to the transferred graft and immune reconstitution post alloHSCT. Due to the variety of immune cells and their distinct roles, a broad evaluation of the immune cellular network is warranted in mobilization and reconstitution studies in alloHSCT. Here, we propose a comprehensive phenotypic analysis of 26 immune cell subsets with multicolor flow cytometry from only 100µl whole blood per time point. Using this approach, we provide an extensive longitudinal analysis of almost 200 time points from 21 donor-recipient pairs. We observe a broad mobilization of innate and adaptive immune cell subsets after granulocyte-colony stimulating factor (G-CSF) treatment of healthy donors. Our data suggest that the relative quantitative immune cell subset composition in recipients approaches that of healthy donors from day +180 post alloHSCT onwards. Correlation of donor and recipient cell counts reveals distinct association patterns for different immune cell subsets and hierarchical clustering of recipient cell counts identifies distinct reconstitution groups in the first month after transplantation. We suggest our comprehensive immune subset analysis as a feasible and time efficient approach for a broad immune assessment for future clinical studies in the context of alloHSCT. This comprehensive cell composition assessment can be a critical step towards personalized graft composition strategies and individualized therapy management in areas such as GvHD prophylaxis in the highly complex immunological setting of alloHSCT.
异基因造血干细胞移植（alloHSCT）的成功和并发症与移植后的移植物和 alloHSCT 后的免疫重建密切相关。由于免疫细胞种类繁多，其作用也各不相同，因此在 alloHSCT 的动员和重建研究中，有必要对免疫细胞网络进行广泛评估。在这里，我们提出了一种使用多色流式细胞术对 26 种免疫细胞亚群进行综合表型分析的方法，每次仅需 100µl 全血。使用这种方法，我们对 21 对供体-受体对的近 200 个时间点进行了广泛的纵向分析。我们观察到健康供体在粒细胞集落刺激因子（G-CSF）治疗后，固有和适应性免疫细胞亚群广泛动员。我们的数据表明，受体中相对定量的免疫细胞亚群组成从 alloHSCT 后第 180 天开始接近健康供体。供体和受体细胞计数的相关性揭示了不同免疫细胞亚群的不同关联模式，而受体细胞计数的层次聚类在移植后第一个月确定了不同的重建组。我们建议将我们的全面免疫亚群分析作为 alloHSCT 背景下未来临床研究中广泛免疫评估的可行且高效的方法。这种全面的细胞组成评估可以成为个性化移植物组成策略和个体化治疗管理的关键步骤，例如 alloHSCT 中高度复杂的免疫环境中的 GvHD 预防。
