The full neutrophil heterogeneity and differentiation landscape remains incompletely characterized. Here, we profiled >25,000 differentiating and mature mouse neutrophils using single-cell RNA sequencing to provide a comprehensive transcriptional landscape of neutrophil maturation, function and fate decision in their steady state and during bacterial infection. Eight neutrophil populations were defined by distinct molecular signatures. The three mature peripheral blood neutrophil subsets arise from distinct maturing bone marrow neutrophil subsets. Driven by both known and uncharacterized transcription factors, neutrophils gradually acquire microbicidal capability as they traverse the transcriptional landscape, representing an evolved mechanism for fine-tuned regulation of an effective but balanced neutrophil response. Bacterial infection reprograms the genetic architecture of neutrophil populations, alters dynamic transitions between subpopulations and primes neutrophils for augmented functionality without affecting overall heterogeneity. In summary, these data establish a reference model and general framework for studying neutrophil-related disease mechanisms, biomarkers and therapeutic targets at single-cell resolution.

完整的中性粒细胞异质性和分化情况仍然不完整。在这里，我们使用单细胞RNA测序分析了超过25,000个分化的和成熟的小鼠嗜中性粒细胞，以提供嗜中性粒细胞在其稳态和细菌感染过程中的成熟，功能和命运决定的全面转录情况。通过不同的分子特征定义了八个嗜中性粒细胞群。三个成熟的外周血中性粒细胞亚群来自不同的成熟骨髓中性粒细胞亚群。在已知和未表征的转录因子的驱动下，嗜中性粒细胞在穿越转录环境时逐渐获得杀微生物能力，这代表了对有效但平衡的嗜中性粒细胞应答进行精细调节的进化机制。细菌感染可重编程中性粒细胞种群的遗传结构，改变亚种群和原始中性粒细胞之间的动态过渡，以增强功能，而不会影响整体异质性。总而言之，这些数据建立了一个参考模型和总体框架，用于以单细胞分辨率研究嗜中性粒细胞相关疾病的机制，生物标志物和治疗靶标。