Peanut oral immunotherapy transiently expands circulating Ara h 2-specific B cells with a homologous repertoire in unrelated subjects
Peanut oral immunotherapy transiently expands circulating Ara h 2-specific B cells with a homologous repertoire in unrelated subjects
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DOI：
10.1016/j.jaci.2015.03.026
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发表时间：
2015-07-01
影响因子：
14.2
通讯作者：
Shreffler, Wayne G.
中科院分区：
文献类型：
--
作者：
Patil, Sarita U.;Ogunniyi, Adebola O.;Shreffler, Wayne G.
背景资料：花生口服免疫疗法（PNOIT）在一部分患者中诱导对花生的持久耐受，并诱导可能在临床保护中发挥作用的特异性抗体。然而，诱导的抗体克隆PNOIT临床tolerance的贡献是unknown.Objective：我们假设PNOIT诱导克隆，过敏原特异性B细胞反应，可以作为临床outcomes.Methods的替代：我们使用了荧光Ara h 2多聚体的亲和选择Ara h 2特异性B细胞和随后的单细胞免疫球蛋白扩增。相关克隆的多样性通过来自循环记忆B细胞的免疫球蛋白重链的下一代测序进行评估，其中在Illumina MiSeq平台上进行2x250配对末端测序。PNOIT诱导循环Ara h 2特异性记忆B细胞的早期和短暂扩增，其在第7周达到峰值。来自记忆细胞的arah 2特异性序列具有与亲和力成熟一致的非沉默突变率。Ara h 2特异性抗体库是寡克隆的。循环记忆B细胞的下一代基于测序的库分析揭示了3个无关受试者中相关序列的会聚选择的证据，表明存在相似的Ara h 2特异性B细胞克隆。使用新的亲和选择方法来鉴定抗原特异性B细胞，我们证明了早期PNOIT诱导的Ara h 2特异性B-细胞受体库是寡克隆的和体细胞超突变的，并且在与会聚选择一致的无关受试者中共享相似的克隆组。
Background: Peanut oral immunotherapy (PNOIT) induces persistent tolerance to peanut in a subset of patients and induces specific antibodies that might play a role in clinical protection. However, the contribution of induced antibody clones to clinical tolerance in PNOIT is unknown.Objective: We hypothesized that PNOIT induces a clonal, allergen-specific B-cell response that could serve as a surrogate for clinical outcomes.Methods: We used a fluorescent Ara h 2 multimer for affinity selection of Ara h 2-specific B cells and subsequent single-cell immunoglobulin amplification. The diversity of related clones was evaluated by means of next-generation sequencing of immunoglobulin heavy chains from circulating memory B cells with 2x250 paired-end sequencing on the Illumina MiSeq platform.Results: Expression of class-switched antibodies from Ara h 2-positive cells confirms enrichment for Ara h 2 specificity. PNOIT induces an early and transient expansion of circulating Ara h 2-specific memory B cells that peaks at week 7. Ara h 2-specific sequences from memory cells have rates of nonsilent mutations consistent with affinity maturation. The repertoire of Ara h 2-specific antibodies is oligoclonal. Next-generation sequencing-based repertoire analysis of circulating memory B cells reveals evidence for convergent selection of related sequences in 3 unrelated subjects, suggesting the presence of similar Ara h 2-specific B-cell clones.Conclusions: Using a novel affinity selection approach to identify antigen-specific B cells, we demonstrate that the early PNOIT-induced Ara h 2-specific B-cell receptor repertoire is oligoclonal and somatically hypermutated and shares similar clonal groups among unrelated subjects consistent with convergent selection.