Iinuma Tomohisa, Kiuchi Masahiro, Hirahara Kiyoshi, Kurita Junya, Kokubo Kota, Yagyu Hiroyuki, Yoneda Riyo, Arai Tomoyuki, Sonobe Yuri, Fukuyo Masaki, Kaneda Atsushi, Yonekura Syuji, Nakayama Toshinori, Okamoto Yoshitaka, Hanazawa Toyoyuki
Department of Otorhinolaryngology, Head and Neck Surgery, Chiba University Graduate School of Medicine, Chiba, Japan.
Department of Immunology, Chiba University Graduate School of Medicine, Chiba, Japan.
J Allergy Clin Immunol. 2022 Oct;150(4):850-860.e5. doi: 10.1016/j.jaci.2022.06.024. Epub 2022 Jul 19.
Allergic rhinitis is a growing problem worldwide. Currently the only treatment that can modify the disease is antigen-specific immunotherapy, but its mechanism of action is not fully understood.
We comprehensively investigated the role and changes of antigen-specific T cells before and after sublingual immunotherapy (SLIT) for Japanese cedar pollinosis.
We cultured peripheral blood mononuclear cells obtained both before and 1 year after initiating SLIT and used a combination of single-cell RNA sequencing and repertoire sequencing. To investigate biomarkers, we used cells from patients participating a phase 2/3 trial of SLIT tablets for Japanese cedar pollinosis and cells from outpatients with good and poor response.
Antigen-stimulated culturing after SLIT led to clonal expansion of T2 and regulatory T cells, and most of these CD4 T cells retained their CDR3 regions before and after treatment, indicating antigen-specific clonal responses and differentiation resulting from SLIT. However, SLIT reduced the number of clonal functional T2 cells but increased the trans-type T2 cell population that expresses musculin (MSC), TGF-β, and IL-2. Trajectory analysis suggested that SLIT induced clonal differentiation of the trans-type T2 cells differentiated into regulatory T cells. Using real-time PCR, we found that the MSC levels increased in the active SLIT group and those with good response after 1 year of treatment.
The combination of single-cell RNA sequencing and repertoire analysis helped reveal part of the underlying mechanism: SLIT promotes the expression of MSC on pathogenic T2 cells and suppresses their function. MSC may be a potential biomarker of SLIT for allergic rhinitis.
变应性鼻炎在全球范围内是一个日益严重的问题。目前，唯一能改变该病的治疗方法是抗原特异性免疫疗法，但其作用机制尚未完全明确。
我们全面研究了日本雪松花粉症患者舌下免疫治疗（SLIT）前后抗原特异性T细胞的作用及变化。
我们培养了开始SLIT前及1年后获取的外周血单个核细胞，并结合使用单细胞RNA测序和谱系测序。为了研究生物标志物，我们使用了参与日本雪松花粉症SLIT片剂2/3期试验患者的细胞以及反应良好和反应不佳的门诊患者的细胞。
SLIT后抗原刺激培养导致T2细胞和调节性T细胞克隆性扩增，并且这些CD4 T细胞中的大多数在治疗前后保留了其互补决定区3（CDR3）区域，表明SLIT导致了抗原特异性克隆反应和分化。然而，SLIT减少了克隆功能性T2细胞的数量，但增加了表达肌动蛋白（MSC）、转化生长因子-β（TGF-β）和白细胞介素-2（IL-2）的跨类型T2细胞群体。轨迹分析表明，SLIT诱导跨类型T2细胞克隆分化为调节性T细胞。使用实时聚合酶链反应（PCR），我们发现活性SLIT组和治疗1年后反应良好的患者中MSC水平升高。
单细胞RNA测序和谱系分析相结合有助于揭示部分潜在机制：SLIT促进致病性T2细胞上MSC的表达并抑制其功能。MSC可能是变应性鼻炎SLIT的潜在生物标志物。
