Background/Aims Previous studies reveal that immune-mediated neuroinflammation plays a key role in the etiology of esophageal achalasia. However, the understanding of leucocyte phenotype and proportion is limited. This study aim to evaluate the phenotypes of leukocytes and peripheral blood mononuclear cells transcriptomes in esophageal achalasia. Methods We performed high-dimensional flow cytometry to identified subsets of peripheral leukocytes, and further validated in lower esophageal sphincter histologically. RNA sequencing was applied to investigate the transcriptional changes in peripheral blood mononuclear cells of patients with achalasia. Cell-type Identification by Estimating Relative Subsets of RNA Transcripts (CIBERSORT) was used for estimating the immune cell types. A differential gene expression analysis was performed and the differential expressed genes were subjected to gene ontology, Kyoto Encyclopedia of Genes and Genomes network, protein-protein interaction network construction. Results An imbalance between innate and adaptive immune cells occurred in achalasia. Specifically, neutrophils and CD8+ T cells increased both in peripheral blood and lower esophageal sphincter in achalasia. Eosinophils decreased in peripheral blood but massively infiltrated in lower esophageal sphincter. CIBERSORT analysis of peripheral blood mononuclear cells RNA sequencing displayed an increased prevalence of CD8+ T cells. 170 dysregulated genes were identified in achalasia, which were enriched in immune cells migration, immune response, etc. Proton pump inhibitor analysis revealed the intersections and gained 7 hub genes in achalasia, which were IL-6, Toll-like receptor 2, IL-1β, tumor necrosis factor, complement C3, and complement C1q A chain. Conclusion Patients with achalasia exhibited an imbalance of systematic innate and adaptive immunity, which may play an important role in the development of achalasia.

中文翻译：

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食管贲门失弛缓症先天免疫和适应性免疫失衡。

背景/目的 先前的研究表明，免疫介导的神经炎症在食管贲门失弛缓症的病因学中起着关键作用。然而，对白细胞表型和比例的了解是有限的。本研究旨在评估食管贲门失弛缓症中白细胞和外周血单核细胞转录组的表型。方法我们进行高维流式细胞术来鉴定外周白细胞亚群，并在下食管括约肌组织学上进一步验证。应用RNA测序研究贲门失弛缓症患者外周血单个核细胞的转录变化。通过估计 RNA 转录本的相对子集进行细胞类型识别 (CIBERSORT) 用于估计免疫细胞类型。对差异基因进行表达分析，并对差异表达基因进行基因本体论、京都基因与基因组百科全书网络、蛋白质-蛋白质相互作用网络构建。结果贲门失弛缓症发生先天免疫细胞和适应性免疫细胞之间的不平衡。具体而言，失弛缓症患者外周血和下食管括约肌中的中性粒细胞和 CD8+ T 细胞均增加。外周血嗜酸性粒细胞减少，食管下括约肌大量浸润。外周血单核细胞 RNA 测序的 CIBERSORT 分析显示 CD8+ T 细胞的患病率增加。在贲门失弛缓症中鉴定出170个失调基因，这些基因在免疫细胞迁移、免疫反应等方面富集。质子泵抑制剂分析揭示了贲门失弛缓症中的交叉点，获得了7个失弛缓症中枢基因，分别是IL-6、Toll样受体2、IL- 1β、肿瘤坏死因子、补体 C3 和补体 C1q A 链。结论 贲门失弛缓症患者存在系统先天免疫和适应性免疫失衡，这可能在贲门失弛缓症的发生发展中起重要作用。