Abstract：
This study sought to compare the behavior of Treg subsets displaying different coexpression patterns of Neuropilin-1 (Nrp1) and Helios, under the influence of gut stress unrelated to hematopoietic stem cell transplantation, pretransplantation conditioning, and posttransplant gastrointestinal acute graft versus host disease (GI-aGvHD). Host CD4+/CD25hi/Foxp3+ Treg cells, identified by flow cytometry, were isolated from various tissues of mice affected by these stressors. Expression of CD25, CTLA-4, CD39, OX40, integrin-β7, LAG3, TGFβ/LAP, granzyme-A, -B, and interleukin-10 was compared in four Treg subsets displaying Helios or Nrp1 only, both or none. Fluorescence-activated cell sorter-sorted Treg subsets, displaying markers affected in a conditioning- and GI-aGVHD-restricted manner, were further investigated by transcriptome profiling and T-cell suppression assays. We found that conditioning by irradiation greatly diminished the relative frequency of Helios+/Nrp1+ Treg, shifting the balance toward Helios-/Nrp1- Treg in the host. Upregulation of integrin-β7 and OX40 occurred in GI-aGvHD-dependent manner in Helios+/Nrp1+ cells but not in Helios-/Nrp1- Treg. Sorted Treg subsets, confirmed to overexpress Nrp1, Helios, OX40, or integrin-β7, displayed superior immunosuppressive activity and enrichment in activation-related messenger RNA transcripts. Our data suggest that conditioning-induced shrinkage of the Nrp1+/Helios+ Treg subset may contribute to the development of GI-GvHD by impairing gut homing and decreasing the efficiency of Treg-mediated immunosuppression.
摘要：
本研究旨在比较显示Neuropilin-1（Nrp1）和Helios的不同共表达模式的Treg亚群的行为，在与造血干细胞移植无关的肠道应激的影响下，移植前预处理,和移植后胃肠道急性移植物抗宿主病（GI-aGvHD）。宿主CD4+/CD25hi/Foxp3+Treg细胞，通过流式细胞术鉴定，从受这些应激源影响的小鼠的各种组织中分离。CD25、CTLA-4、CD39、OX40、整合素β7、LAG3、TGFβ/LAP的表达,颗粒酶A，-B,和白细胞介素-10在四个仅显示Helios或Nrp1的Treg亚群中进行比较，两者或无。荧光激活细胞分选分选的Treg亚群，以条件和GI-aGVHD限制方式显示受影响的标记，通过转录组分析和T细胞抑制测定进一步研究。我们发现通过辐照调节大大降低了Helios/Nrp1Treg的相对频率，将平衡向宿主中的Helios-/Nrp1-Treg转移。整合素-β7和OX40的上调在Helios/Nrp1细胞中以GI-aGvHD依赖性方式发生，但在Helios-/Nrp1-Treg细胞中未发生。排序的Treg子集，确认过度表达Nrp1，Helios，OX40或整合素β7显示出优异的免疫抑制活性，并富集了与激活相关的信使RNA转录本。我们的数据表明，调节诱导的Nrp1/HeliosTreg亚群的收缩可能通过损害肠道归巢和降低Treg介导的免疫抑制效率来促进GI-GvHD的发展。
