Unraveling transcriptomic and functional changes to immune sensing neuronal ganglia in response to allergy and bacteria

项目摘要 过敏影响着数以百万计的美国人，细菌感染加剧了过敏。尽管主流疗法 抑制对过敏或细菌的免疫反应，这些健康问题继续存在。未被探索的目标 新疗法的关键在于神经监测和免疫调节。然而，这种神经影响是 鲜为人知且看似矛盾的是：过敏时免疫反应会加剧，但对 细菌感染，神经反应减少炎症。这怎么可能发生呢？ 一种可能性是，外周神经免疫感觉器可能会受到以下因素的影响 免疫球蛋白图谱和性别，暗示有性激素参与。我们假设这样的情况 变量影响转录组，从而影响迷走神经和颈动脉小体的兴奋性。目标1中的研究 将对患有哮喘的雄性和雌性小鼠的迷走神经和颈动脉体节进行单细胞RNAseq分析， 肺炎以确定这些神经在每种情况下是如何变化的。然后，我们将评估神经 应用最先进的电生理学技术研究免疫球蛋白刺激对每个神经节的反应 录音技术。这个项目将揭示哪个神经丛(迷走神经和颈动脉体)是占优势的。 感觉神经节参与调节过敏和细菌免疫反应，以及性行为如何影响神经- 免疫信号。AIM 2的研究将利用小鼠的DTR(白喉毒素受体)技术选择性地 消融由TRPV1(瞬时受体电位香草素1)或TH(酪氨酸)驱动DTR表达的细胞 羟基酶)。使用这些转基因小鼠将可以消融迷走神经(TRPV1-DTR)或颈动脉小体(TH-TH)。 将白喉毒素(DTX)直接神经微量注射到迷走神经节或颈动脉体神经节 注射生理盐水作为对照。然后，我们将使小鼠暴露在过敏原或细菌感染下。DTX注射将 选择性地取消神经元功能并揭示哪个神经节是脑内主要的神经免疫感受器 雄性和雌性小鼠的特定情况。 这些数据将：1)确定对过敏或细菌刺激的反应如何改变受体基因的表达 以确定从抗炎到促炎的神经免疫信号的表型转换；2)建立 模特和性别差异，将为进一步调查奠定基础，涉及有针对性的击倒 神经免疫感觉神经节识别功能；3)验证我们的模型和方法，为未来的研究做准备 神经免疫感知在疾病状态中的作用。

Project Summary Allergies affect millions of Americans and are exacerbated by bacterial infection. Despite mainstay therapies that suppress immune responses to allergy or bacteria, these health issues continue to persist. An unexplored target for new therapies lies with neural surveillance and regulation of immunity. However, this neural influence is poorly understood and seemingly paradoxical: immune responses are exacerbated in allergy, but in response to bacterial infection, the neural response reduces inflammation. How might this happen? One possibility is that the peripheral neural-immune sensing apparatus might be altered by such factors as immunoglobulin profiles, and gender, implying the participation of sex hormones. We hypothesize that such variables affect the transcriptome and thereby the excitability of the vagus and carotid bodies. Studies in AIM 1 will use single-cell RNAseq analysis of vagal and carotid body ganglia from male and female mice with asthma, pneumonia to identify how these nerves are changed in response to each condition. Then, we will assess nerve activity in response to immunoglobulin stimuli of each ganglia using state-of-the-art electrophysiological recording techniques. This project will reveal which neural cluster (vagus vs. carotid body) is the predominant sensory ganglia involved in regulating allergic vs. bacterial immune responses and how sex may affect neuro- immune signalling. Studies in AIM 2 will utilize DTR (diphtheria toxin receptor) technology in mice to selectively ablate cells where DTR expression is driven by TRPV1 (transient receptor potential vanilloid 1) or TH (tyrosine hydroxylase). Use of these transgenic mice will allow ablation of either vagal (TRPV1-DTR) or carotid body (TH- DTR) centers by direct neural microinjection of diphtheria toxin (DTX) into vagal or carotid body ganglia using saline injections as control. We will then expose mice to either allergen or bacterial infections. DTX injections will selectively abolish neuronal function and reveal which ganglion is the predominant neuro-immune sensor in the specified condition in male and female mice. These data will: 1) identify how the response to allergic or bacterial stimuli alters receptor gene expression in order to identify the phenotypic switch from anti- to pro-inflammatory neural-immune signalling; 2) establish a model and sex differences that will set the stage for further investigation involving targeted knockdown within neuro-immune sensing ganglia to discern function; 3) validate our models and methods for future investigations into the role of neural-immune sensing in disease states.