Salinas Michael L, Mulakala Bharath Kumar, Davidson Laurie A, Cai James J, Donovan Sharon M, Chapkin Robert S, Yeruva Laxmi
Department of Nutrition, Texas A&M University, College Station, TX, United States; CPRIT Single Cell Data Science Core, Texas A&M University, College Station, TX, United States; Texas A&M AgriLife Institute for Advancing Health Through Agriculture, College Station, TX, United States.
Texas A&M AgriLife Institute for Advancing Health Through Agriculture, College Station, TX, United States; Microbiome and Metabolism Research Unit, USDA-ARS, SEA, Little Rock, AR, United States; Arkansas Children's Nutrition Center, Little Rock, AR, United States.
Am J Clin Nutr. 2025 Jul;122(1):196-207. doi: 10.1016/j.ajcnut.2025.04.024. Epub 2025 Apr 26.
Several studies have indicated the benefits of human milk feeding to infants however, mechanisms behind positive health outcomes have not been investigated.
The study aimed to characterize circulating immune cell subpopulation gene expression in human milk-fed (HMF) compared with cow milk formula-fed (FF) infants using single-cell transcriptomics.
Peripheral blood mononuclear cells (PBMCs) were isolated from healthy HMF (n = 6), and FF (n = 3) infants who were 3-3.5 mo old and enrolled in a nonrandomized clinical trial. Single-cell RNA sequencing was used to generate a PBMC atlas and evaluate gene expression in immune cell subsets. Differential expression analysis was performed on each cell type independently after clustering the cells by similar marker gene expression using the scGEAToolbox. Differentially expressed genes were subjected to pathway analyses using an online functional enrichment analysis program.
The relative abundance (%) of T and B lymphocytes, natural killer (NK) cells, and plasmacytoid dendritic cells were similar, whereas monocytes were higher in FF infants than in HMF infants (22.6 ± 10.7 compared with 8.3 ± 5.6; P = 0.0314). In addition, innate and adaptive immune cells from FF infants exhibited a higher activation state compared with HMF infants. We identified 16 distinct cell subsets from the major immune cell types: 3 monocyte subsets, 4 NK subsets, 2 B cell subsets, and 7 T cell subsets. Transcriptional profiles of each peripheral innate and adaptive immune cell subtype varied between HMF and FF infants. Pathway enrichment analysis of cell-specific transcriptional changes within subsets of major cell types revealed that the interleukin (IL)-4/IL-13 signaling pathways were upregulated in FF infants relative to HMF infants.
These findings suggest that human milk downregulates peripheral immune cell cytokine transcriptional signatures linked to allergic inflammation and infection relative to formula feeding.
多项研究表明母乳喂养对婴儿有益，然而，健康有益结果背后的机制尚未得到研究。
本研究旨在通过单细胞转录组学，对母乳喂养（HMF）婴儿与配方奶喂养（FF）婴儿的循环免疫细胞亚群基因表达进行特征分析。
从3至3.5月龄、参加一项非随机临床试验的健康HMF（n = 6）和FF（n = 3）婴儿中分离外周血单核细胞（PBMC）。使用单细胞RNA测序生成PBMC图谱，并评估免疫细胞亚群中的基因表达。在使用scGEAToolbox根据相似的标记基因表达对细胞进行聚类后，对每种细胞类型独立进行差异表达分析。使用在线功能富集分析程序对差异表达基因进行通路分析。
T淋巴细胞、B淋巴细胞、自然杀伤（NK）细胞和浆细胞样树突状细胞的相对丰度（%）相似，而FF喂养婴儿的单核细胞高于HMF喂养婴儿（分别为22.6±10.7与8.3±5.6；P = 0.0314）。此外，与HMF喂养婴儿相比，FF喂养婴儿的先天性和适应性免疫细胞表现出更高的激活状态。我们从主要免疫细胞类型中鉴定出16个不同的细胞亚群：3个单核细胞亚群、4个NK亚群、2个B细胞亚群和7个T细胞亚群。HMF和FF喂养婴儿的每种外周先天性和适应性免疫细胞亚型的转录谱各不相同。主要细胞类型亚群内细胞特异性转录变化的通路富集分析表明，相对于HMF喂养婴儿，FF喂养婴儿的白细胞介素（IL）-4/IL-13信号通路上调。
这些发现表明，相对于配方奶喂养，母乳可下调与过敏性炎症和感染相关的外周免疫细胞细胞因子转录特征。
