Single-cell profiling of peanut-responsive T cells in patients with peanut allergy reveals heterogeneous effector TH2 subsets
Single-cell profiling of peanut-responsive T cells in patients with peanut allergy reveals heterogeneous effector TH2 subsets
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DOI：
10.1016/j.jaci.2017.11.060
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发表时间：
2018-06-01
影响因子：
14.2
通讯作者：
Berin, M. Cecilia
中科院分区：
文献类型：
--
作者：
Chiang, David;Chen, Xintong;Berin, M. Cecilia
背景资料：花生特异性T细胞的表型变异对花生过敏或耐受性的贡献尚不清楚。目的：我们的目的是全面表型花生特异性T细胞在受试者外周血与花生过敏（PA）。方法：我们从PA患者中获得样本，包括一组进行免疫治疗试验的基线花生攻击（Consortium of Food Allergy Research [CoFAR] 6）。受试者被证实患有PA，或者如果他们通过了1克花生的挑战，他们被称为高阈值受试者。还招募了健康对照（HC）受试者。在用花生提取物刺激6至18小时后，基于CD 154表达鉴定花生应答性T细胞。通过流式细胞术和单细胞RNA测序对细胞进行分析。结果：PA患者具有组织和滤泡归巢花生反应性CD 4 1 T细胞，具有TH 2分化的异质模式，而对照受试者对花生的T细胞反应检测不到。PA组在表达调节性T（Treg）细胞标志物的细胞上具有延迟的和IL-2依赖性的CD 154上调，这在HC或高阈值受试者中不存在。Treg细胞的耗竭增强了HC受试者和PA患者体外细胞因子的产生，但与高度分化的T（H）2细胞相关的细胞因子对PA患者的Treg细胞抑制更具抵抗力。通过单细胞RNA测序分析基因表达，鉴定出IL 4、IL 5、IL 9、IL 13和IL-25受体IL 17 RB表达高度相关的T细胞。结论：PA患者存在高度分化的T（H）2细胞，产生T（H）2相关细胞因子，其功能超出IgE类转换。在花生反应性T细胞中，多功能T（H）2反应比Treg细胞缺陷更明显。
Background: The contribution of phenotypic variation of peanut-specific T cells to clinical allergy or tolerance to peanut is not well understood.Objectives: Our objective was to comprehensively phenotype peanut-specific T cells in the peripheral blood of subjects with and without peanut allergy (PA).Methods: We obtained samples from patients with PA, including a cohort undergoing baseline peanut challenges for an immunotherapy trial (Consortium of Food Allergy Research [CoFAR] 6). Subjectswere confirmed as having PA, or if they passed a 1-g peanut challenge, they were termed high-threshold subjects. Healthy control (HC) subjects were also recruited. PeanutresponsiveTcellswere identified based onCD154 expression after 6 to 18 hours of stimulation with peanut extract. Cells were analyzed by using flow cytometry and single-cell RNA sequencing.Results: Patients with PA had tissue-and follicle-homing peanutresponsive CD4 1 T cells with a heterogeneous pattern of TH2 differentiation, whereas control subjects had undetectable T-cell responses to peanut. The PA group had a delayed and IL-2-dependent upregulation of CD154 on cells expressing regulatory T (Treg) cell markers, which was absent in HC or high-threshold subjects. Depletion of Treg cells enhanced cytokine production in HC subjects and patients with PA in vitro, but cytokines associated with highly differentiated T(H)2 cells were more resistant to Treg cell suppression in patients with PA. Analysis of gene expression by means of single-cell RNA sequencing identified T cells with highly correlated expression of IL4, IL5, IL9, IL13, and the IL-25 receptor IL17RB.Conclusions: These results demonstrate the presence of highly differentiated T(H)2 cells producing T(H)2-associated cytokines with functions beyond IgE class-switching in patients with PA. A multifunctional T(H)2 response was more evident than a Treg cell deficit among peanut-responsive T cells.