Willem Alexander Children's Hospital, Laboratory for Pediatric Immunology, Department of Pediatrics, Leiden University Medical Center, 2300 RC Leiden, The Netherlands.
Willem Alexander Children's Hospital, Division of Neonatology, Department of Pediatrics, Leiden University Medical Center, 2300 RC Leiden, The Netherlands.
Int J Mol Sci. 2023 Jan 25;24(3):2372. doi: 10.3390/ijms24032372.
Epigenetic immune cell counting is a DNA (de)methylation-based technique which can be used to quantify lymphocyte subsets on dried blood spots (DBS). The foregoing techniques allow for a retrospective investigation of immune cell profiles in newborns. In this study, we used this technique for determining lymphocyte subcounts as a potential biomarker for necrotizing enterocolitis (NEC). We investigated whether this technique can be implemented in the field of neonatology, by testing whether regulatory T cell (Treg) levels are pre-existently low in preterms with NEC. Newborn screening (NBS) cards from 32 preterms with NEC and 32 age- and weight-matched preterm controls, and 60 healthy term newborns, were analyzed. Relative and absolute cell counts were determined for CD3+, CD4+, CD8+, Th17, and Treg T cells. For both relative and absolute cell counts of CD3+, CD4+, CD8+, and Th17 T cells, significant differences were found between healthy term controls and both preterm groups, but not between preterm groups. For Tregs, no significant differences were found in either relative or absolute counts between any of the newborn groups. This study demonstrates the principle of epigenetic immune cell counting to analyze lymphocyte subsets in preterm neonates.
基于 DNA（去）甲基化的表观遗传免疫细胞计数技术可用于定量分析干血斑（DBS）中的淋巴细胞亚群。上述技术可用于回顾性研究新生儿的免疫细胞谱。在这项研究中，我们使用该技术来确定淋巴细胞亚群计数，作为坏死性小肠结肠炎（NEC）的潜在生物标志物。我们通过检测 NEC 早产儿中调节性 T 细胞（Treg）水平是否预先降低，来研究该技术是否可以在新生儿领域实施。分析了 32 例 NEC 早产儿、32 例年龄和体重匹配的早产儿对照和 60 例健康足月新生儿的新生儿筛查（NBS）卡。测定 CD3+、CD4+、CD8+、Th17 和 Treg T 细胞的相对和绝对细胞计数。对于 CD3+、CD4+、CD8+和 Th17 T 细胞的相对和绝对细胞计数，健康足月对照组与早产儿组均存在显著差异，但早产儿组之间无差异。对于 Tregs，任何新生儿组的相对或绝对计数均无显著差异。本研究证明了表观遗传免疫细胞计数分析早产儿淋巴细胞亚群的原理。
