Department of Pediatric Pneumology, Allergy and Neonatology, Hannover Medical School, Hannover, Germany.
Department of Pediatric Pneumology, Allergy and Neonatology, Hannover Medical School, Hannover, Germany.
J Allergy Clin Immunol. 2014 Jan;133(1):172-80.e1-10. doi: 10.1016/j.jaci.2013.06.047. Epub 2013 Sep 5.
Primary antibody deficiencies represent the most prevalent, although very heterogeneous, group of inborn immunodeficiencies, with a puzzling complexity of cellular and molecular processes involved in disease pathogenesis.
We aimed to study in detail the kinetics of CD40 ligand/IL-21-induced B-cell differentiation to define new biomarker sets for further research into primary antibody deficiencies.
We applied high-content screening methods to monitor B-cell activation on the cellular (chip cytometry) and transcriptomic (RNA microarray) levels.
The complete activation process, including stepwise changes in protein and RNA expression patterns, entry into the cell cycle, proliferation and expression of activation-induced cytidine deaminase (AID), DNA repair enzymes, and post-class-switch expression of IgA and IgG, was successfully monitored during in vitro differentiation. We identified a number of unknown pathways engaged during B-cell activation, such as CXCL9/CXCL10 secretion by B cells. Finally, we evaluated a deduced set of biomarkers on a group of 18 patients with putative or proved intrinsic B-cell defects recruited from the European Society for Immunodeficiencies database and successfully predicted 2 AID defects and 1 DNA repair defect. Complete absence of class-switched B cells was a sensitive predictor of AID deficiency and should be further evaluated as a diagnostic biomarker.
The biomarkers found in this study could be used to further study the complex process of B-cell activation and to understand conditions that lead to the development of primary antibody deficiencies.
原发性抗体缺陷是最常见的，尽管非常异质，一组先天免疫缺陷，涉及疾病发病机制的细胞和分子过程令人费解的复杂性。
我们旨在详细研究 CD40 配体/IL-21 诱导的 B 细胞分化的动力学，以确定新的生物标志物集，用于进一步研究原发性抗体缺陷。
我们应用高内涵筛选方法在细胞（芯片细胞术）和转录组（RNA 微阵列）水平上监测 B 细胞的激活。
成功监测了体外分化过程中完整的激活过程，包括蛋白质和 RNA 表达模式的逐步变化、进入细胞周期、增殖和激活诱导胞苷脱氨酶（AID）、DNA 修复酶的表达以及 IgA 和 IgG 的后类别转换表达。我们发现了一些在 B 细胞激活过程中涉及的未知途径，例如 B 细胞分泌的 CXCL9/CXCL10。最后，我们在从欧洲免疫缺陷学会数据库招募的 18 名疑似或证实存在内在 B 细胞缺陷的患者组上评估了一组推断的生物标志物，并成功预测了 2 例 AID 缺陷和 1 例 DNA 修复缺陷。完全缺乏类别转换的 B 细胞是 AID 缺陷的敏感预测因子，应进一步作为诊断生物标志物进行评估。
本研究中发现的生物标志物可用于进一步研究 B 细胞激活的复杂过程，并了解导致原发性抗体缺陷发展的条件。
