Abstract：
The early diagnosis of neonatal sepsis is crucial as it remains a prevalent cause of neonatal mortality. In this study, we conducted an analysis on the clinical data and detection indicators of 22 cases with sepsis and 62 cases without sepsis among neonates. Our findings indicate that the clinical signs observed in neonates with sepsis lack specificity. In addition, the commonly used clinical inflammatory indicators (such as leukocyte count, neutrophil-to-lymphocyte ratio [NLR], C-reactive protein [CRP], procalcitonin) exhibit limited sensitivity and specificity. Furthermore, the current clinical measures lack the assessment of inflammatory factors. Therefore, in order to enhance the accuracy of early sepsis diagnosis in neonates, we have employed a novel microfluidic-based single-cell technology platform for the analysis of 32 cytokines secreted by neutrophils at the individual cell level under various toxin stimulation conditions. We have further investigated and compared the disparities in single-cell protein secretomics between umbilical cord blood neutrophils and healthy adult peripheral neutrophils within an in vitro sepsis model. Our findings indicate that in a resting state UCB neutrophils exhibited lower polyfunctionality compared with healthy adult blood neutrophils, and notable variations in cytokine secretion profiles were detected between the two groups. However, the polyfunctionality of UCB neutrophils significantly increased and surpassed that of healthy adult neutrophils when exposed to alpha-hemolysin or lipopolysaccharide. UCB neutrophils secreted a wide range of chemokines and inflammatory factors, among which GM-CSF and IL-18 were the most significant. Furthermore, we initially categorized the functional subgroups of neutrophils by considering the secretion of five primary cytokines by neutrophils (GM-CSF, IL-18, IL-8, MIP-1β, and MIF). The current study, for the first time, examined in detail the heterogeneity of protein secretion and the functional diversity of UCB neutrophils stimulated by different antigens. Moreover, new insight into neonatal sepsis, early diagnosis, and wider clinical applications of UCB neutrophils are provided by these data.
摘要：
新生儿败血症的早期诊断至关重要，因为它仍然是新生儿死亡的主要原因。在这项研究中,对新生儿败血症22例和无败血症62例的临床资料及检测指标进行分析。我们的发现表明，在败血症新生儿中观察到的临床体征缺乏特异性。此外,常用的临床炎症指标(如白细胞计数、中性粒细胞与淋巴细胞比率[NLR]，C反应蛋白[CRP]，降钙素原）表现出有限的敏感性和特异性。此外，目前的临床措施缺乏对炎症因子的评估。因此,为了提高新生儿败血症早期诊断的准确性,我们采用了一种新型的基于微流控的单细胞技术平台,在各种毒素刺激条件下,在单个细胞水平分析中性粒细胞分泌的32种细胞因子.我们进一步研究并比较了体外脓毒症模型中脐带血中性粒细胞和健康成人外周中性粒细胞在单细胞蛋白分泌组学中的差异。我们的研究结果表明，在静息状态下，UCB中性粒细胞表现出较低的多功能与健康成人血液中性粒细胞相比，两组之间检测到细胞因子分泌谱的显著差异。然而,当暴露于α-溶血素或脂多糖时,UCB中性粒细胞的多功能性显著增加并超过健康成人中性粒细胞。UCB中性粒细胞分泌广泛的趋化因子和炎症因子，其中GM-CSF和IL-18最为显著。此外，我们最初通过考虑中性粒细胞分泌五种主要细胞因子（GM-CSF，IL-18，IL-8，MIP-1β，和MIF）。目前的研究,第一次,详细检查了蛋白质分泌的异质性和不同抗原刺激的UCB中性粒细胞的功能多样性。此外，对新生儿败血症的新见解，早期诊断,这些数据提供了UCB中性粒细胞的更广泛的临床应用。
