Pediatric Department A and the Immunology Service, Jeffrey Modell Foundation Center, Edmond and Lily Safra Children's Hospital, Sheba Medical Center, 52621, Tel HaShomer, Israel.
Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.
J Clin Immunol. 2023 Jan;43(1):109-122. doi: 10.1007/s10875-022-01349-8. Epub 2022 Aug 31.
Patients with Wiskott-Aldrich syndrome (WAS) harbor mutations in the WAS gene and suffer from immunodeficiency, microthrombocytopenia, and eczema. T-cells play an important role in immune response in the skin and the γδT-cells have an important role in skin homeostasis. Since WAS patients often present with eczema, we wanted to examine whether the T-cell receptor gamma (TRG) repertoire of the γδT-cells is affected in these patients. In addition, the immunoglobulin heavy chain (IGH) repertoire from genomic DNA of WAS patients was not yet studied. Thus, we sought to determine the effects that specific WAS mutations from our patients have in shaping the TRG and IGH immune repertoires. We collected clinical and genetic data on four WAS patients, each harboring a different mutation in the WAS gene. Using next-generation sequencing (NGS), we analyzed their TRG and IGH repertoires using genomic DNA isolated from their peripheral blood. We analyzed the TRG and IGH repertoire sequences to show repertoire restriction, clonal expansions, preferential utilization of specific V genes, and unique characteristics of the antigen binding region in WAS patients with eczema compared to healthy controls. Both the TRG and IGH repertoire showed diverse repertoire comparable to healthy controls on one the hand, and on the other hand, the IGH repertoire showed increased diversity, more evenly distributed repertoire and immaturity of the antigen binding region. Thus, we demonstrate by analyzing the repertoire based on genomic DNA, the various effect that WAS mutations have in shaping the TRG and IGH adaptive immune repertoires.
患有威特综合征（Wiskott-Aldrich syndrome，WAS）的患者存在 WAS 基因突变，表现为免疫缺陷、血小板减少和湿疹。T 细胞在皮肤免疫反应中发挥重要作用，γδT 细胞在皮肤稳态中发挥重要作用。由于 WAS 患者常出现湿疹，我们想研究这些患者的 γδT 细胞的 T 细胞受体 γ（TRG）库是否受到影响。此外，WAS 患者的免疫球蛋白重链（immunoglobulin heavy chain，IGH）库尚未研究。因此，我们试图确定来自我们患者的特定 WAS 突变对 TRG 和 IGH 免疫库的影响。我们收集了四名 WAS 患者的临床和遗传数据，每位患者的 WAS 基因都存在不同的突变。使用下一代测序（next-generation sequencing，NGS），我们使用从外周血中分离的基因组 DNA 分析了他们的 TRG 和 IGH 库。我们分析了 TRG 和 IGH 库序列，以显示在患有湿疹的 WAS 患者与健康对照者相比，库的限制、克隆扩增、特定 V 基因的优先利用以及抗原结合区的独特特征。一方面，TRG 和 IGH 库与健康对照者的多样性库相当，另一方面，IGH 库显示出多样性增加、更均匀分布的库和抗原结合区的不成熟。因此，我们通过分析基于基因组 DNA 的库来证明，WAS 突变在塑造 TRG 和 IGH 适应性免疫库方面具有各种影响。
