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Tracking immunodynamics by identification of S-G2/M-phase T cells in human peripheral blood.
Journal of Autoimmunity ( IF 12.8 ) Pub Date : 2020-05-12 , DOI: 10.1016/j.jaut.2020.102466
Miguel Muñoz-Ruiz 1 , Irma Pujol-Autonell 2 , Hefin Rhys 3 , Heather M Long 4 , Maria Greco 5 , Mark Peakman 2 , Tim Tree 2 , Adrian C Hayday 6 , Francesca Di Rosa 7
Affiliation  

The ready availability of human blood makes it the first choice for immuno-monitoring. However, this has been largely confined to static metrics, particularly resting T cell phenotypes. Conversely, dynamic assessments have mostly relied on cell stimulation in vitro which is subject to multiple variables. Here, immunodynamic insights from the peripheral blood are shown to be obtainable by applying a revised approach to cell-cycle analysis. Specifically, refined flow cytometric protocols were employed, assuring the reliable quantification of T cells in the S-G2/M phases of the cell-cycle (collectively termed “T Double S” for T cells in S-phase in Sanguine: in short “TDS” cells). Without protocol refinement, TDS could be either missed, as most of them layed out of the conventional lymphocyte gates, or confused with cell doublets artefactually displaying high DNA-content. To illustrate the nature of TDS cells, and their relationship to different immunodynamic scenarios, we examined them in healthy donors (HD); infectious mononucleosis (IM) patients versus asymptomatic EBV+ carriers; and recently-diagnosed T1D patients. TDS were reproducibly more abundant among CD8+ T cells and a defined subset of T-regulatory CD4+ T cells, and were substantially increased in IM and a subset of T1D patients. Of note, islet antigen-reactive TDS cell frequencies were associated with an aggressive T cell effector phenotype, suggesting that peripheral blood can reflect immune events within tissues in T1D, and possibly in other organ-specific autoimmune diseases.

Our results suggest that tracking TDS cells may provide a widely applicable means of gaining insight into ongoing immune response dynamics in a variety of settings, including tissue immunopathologies where the peripheral blood has often not been considered insightful.



中文翻译:

通过鉴定人外周血中的 S-G2/M 期 T 细胞来跟踪免疫动力学。

人血的现成可用性使其成为免疫监测的首选。然而,这在很大程度上仅限于静态指标,特别是静息 T 细胞表型。相反,动态评估主要依赖于受多种变量影响的体外细胞刺激。在这里,外周血的免疫动力学见解显示可以通过将修改后的方法应用于细胞周期分析来获得。具体而言,精流式细胞协议被雇用,保证在SG T细胞的可靠定量2细胞周期(统称为“T双重S”为/ M期Ť细胞在小号-PHASE小号anguine:在短“ Ť DS“ 细胞)。如果不改进方案,T DS可能会被遗漏,因为它们中的大多数都位于传统的淋巴细胞门之外,或者与人为显示高 DNA 含量的细胞双联体混淆。为了说明 T DS细胞的性质以及它们与不同免疫动力学场景的关系,我们在健康供体 (HD) 中检查了它们;传染性单核细胞增多症 (IM) 患者与无症状 EBV +携带者;和最近确诊的 T1D 患者。T DS在 CD8 + T 细胞和特定的 T 调节性 CD4 + T 细胞亚群中可重复地更丰富,并且在 IM 和 T1D 患者亚群中显着增加。值得注意的是,胰岛抗原反应性 T DS 细胞频率与侵袭性 T 细胞效应表型相关,表明外周血可以反映 T1D 组织内的免疫事件,也可能反映其他器官特异性自身免疫疾病。

我们的结果表明,跟踪 T DS细胞可能提供一种广泛适用的方法,可以深入了解各种环境中正在进行的免疫反应动态,包括外周血通常不被认为具有洞察力的组织免疫病理学。

更新日期:2020-07-20
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