Correlating neutrophil function and plasma cytokine profiles with progression of ME/CFS and Long-Covid/PASC

摘要 肌痛性脑炎/慢性疲劳综合征(ME/CFS)是一种复杂的慢性疾病，与 长时间的炎症信号。长期冠状病毒/急性冠状病毒后遗症(PASC)已被认为是一种 相对常见的SARS-CoV-2感染结局，似乎反映了ME/CFS的许多特征， 这表明这些疾病的病因可能是相同的[1，2]。中性粒细胞是最常见的免疫 细胞在循环中，并且对炎症信号的变化非常敏感。我们已经开发出一种 独特的微流控分析小组，可以从一滴新鲜血液中探测中性粒细胞的功能。我们最近用了 对6例ICC确诊的ME/CFS患者和1例PASC患者的新鲜指血进行检测 并发现初步证据表明，ME/CFS的多种中性粒细胞功能相对于 疾病的发展。引人注目的是，PASC受试者中性粒细胞功能的变化与 早期ME/CFS。我们项目的目标是使用这些微流控分析来探索中性粒细胞的功能。 ICC-ME/CFS、PASC受试者和健康对照组的更大队列。我们将描述中性粒细胞的运动性 以及从一滴新鲜血液中形成细胞外陷阱(NETsis)，无论是在基线还是在体外 刺激，在所有受试者中。我们还将使用一个已建立的小组来表征促炎症细胞因子在 在中性粒细胞点刺的同时采集ME/CFS、PASC和健康对照组受试者的血液 样本。这将决定促炎细胞因子负荷是否与中性粒细胞运动性和基线相关 跨组蚊虫感染。该提案的一个主要目标是分析中性粒细胞的反应作为 ME/CFS和PASC的病程。相对较新发病的中性粒细胞行为特征 PASC将为未来对这种情况的纵向研究奠定基础，并将有助于确定 机械上与ME/CFS重叠或不存在。

SUMMARY Myalgic Encephalitis/Chronic Fatigue Syndrome (ME/CFS) is a complex chronic condition associated with prolonged inflammatory signaling. Long Covid/Post-Acute Sequelae of Covid (PASC) has been noted as a relatively common outcome of SARS-CoV-2 infection, and appears to mirror many of the features of ME/CFS, suggesting that the etiology of these conditions might be shared [1, 2]. Neutrophils are the most common immune cells in the circulation and are exquisitely sensitive to changes in inflammatory signaling. We have developed a unique panel of microfluidic assays that probe neutrophil function from a drop of fresh blood. We recently used these assays on fresh fingerpick blood obtained from 6 ICC-diagnosed ME/CFS subjects and 1 PASC subject and found preliminary evidence that multiple neutrophil functions were altered in ME/CFS relative to progression of disease. Strikingly, the PASC subject exhibited changes in neutrophil function most similar to early-stage ME/CFS. The goal of our project is to use these microfluidic assays to probe neutrophil function in a larger cohort of ICC-ME/CFS, PASC subjects, and healthy controls. We will characterize neutrophil motility and extracellular trap formation (NETosis) from a drop of fresh blood, both at baseline and following in vitro stimulation, in all subjects. We will also use an established panel to characterize proinflammatory cytokines in ME/CFS, PASC, and healthy control subject blood collected at the same time as the neutrophil fingerprick samples. This will determine if proinflammatory cytokine burden will correlate with neutrophil motility and baseline NETosis across groups. An overarching aim of the proposal is to analyze neutrophil responses as a function of illness duration in both ME/CFS and PASC. Characterization of neutrophil behavior in relatively recent-onset PASC will lay the groundwork for future longitudinal studies of this condition and will help determine the mechanistic overlap with ME/CFS or lack thereof.