Domingo-Vila Clara, Williams Evangelia D, Smithmyer Megan E, Benson Basilin, Ferrat Lauric A, Arif Sefina, Hudson Michelle, Dobbs Becky, Johnson Matthew B, Yardley Iain, Speake Cate, Oram Richard A, Tree Timothy I M, Blaise Benjamin J
School of Immunology and Microbial Sciences, King's College London, London, United Kingdom.
Center for Interventional Immunology, Benaroya Research Institute, Seattle, WA, United States.
Front Immunol. 2025 Jul 24;16:1618080. doi: 10.3389/fimmu.2025.1618080. eCollection 2025.
Recruiting very young, healthy children to serve as age-matched controls in research presents substantial ethical and practical challenges. One potential approach to address this issue is to recruit healthy children who are referred for elective procedures under general anesthesia. As infants are typically anesthetized using volatile anesthetics before cannula insertion for additional drug administration, blood samples become readily accessible after the onset of drug-induced coma. However, since prolonged exposure to inhaled anesthetic agents is known to have immune-modulating effects that could affect their suitability as experimental controls, we aimed to investigate whether immune changes are also present in samples collected immediately after gas induction in children undergoing elective dental procedures. The composition and transcriptional profile of whole blood immune cells were assessed using multiparameter flow cytometry and bulk RNA-sequencing, respectively. Cryopreserved PBMCs were used to study changes in the phenotype of polyclonally activated CD4 T cells by single-cell RNA sequencing using the 10x Genomics (Pleasanton, CA, USA) platform. We report that inhaled anesthetic induction with a combination of nitrous oxide and sevoflurane has minimal effect on immune system composition and transcriptional profiles, and does not alter the phenotype of CD4 T cells activated with staphylococcal enterotoxin B (SEB). However, we observed increased absolute cell counts in specific leucocyte populations. We conclude that blood samples collected during elective procedures under general anesthesia may represent a valuable opportunity for recruiting healthy children for research studies, depending on the intended assays.
招募非常年幼、健康的儿童作为研究中的年龄匹配对照存在重大的伦理和实际挑战。解决这个问题的一种潜在方法是招募因择期手术接受全身麻醉的健康儿童。由于婴儿通常在插入套管进行额外药物给药之前使用挥发性麻醉剂进行麻醉，在药物诱导昏迷开始后，血液样本很容易获取。然而，由于已知长时间接触吸入麻醉剂具有免疫调节作用，这可能会影响他们作为实验对照的适用性，我们旨在研究在接受择期牙科手术的儿童中，气体诱导后立即采集的样本中是否也存在免疫变化。分别使用多参数流式细胞术和大量RNA测序评估全血免疫细胞的组成和转录谱。使用美国加利福尼亚州普莱森顿的10x Genomics平台，通过单细胞RNA测序，利用冷冻保存的外周血单核细胞（PBMC）研究多克隆激活的CD4 T细胞表型的变化。我们报告，一氧化二氮和七氟醚联合吸入麻醉诱导对免疫系统组成和转录谱的影响最小，并且不会改变用葡萄球菌肠毒素B（SEB）激活的CD4 T细胞的表型。然而，我们观察到特定白细胞群体的绝对细胞计数增加。我们得出结论，根据预期的检测方法，在全身麻醉下的择期手术期间采集的血液样本可能为招募健康儿童进行研究提供一个宝贵的机会。
