Generation of immune-enhancing neutrophils by subclinical super-low dose LPS

We have generated immune-enhancing neutrophils by culturing murine primary bone marrow derived neutrophils with either super-low dose of LPS. Immune-enhancing neutrophils preferentially express co-stimulatory molecules such as CD74, CD44 and CD86, and exhibit reduced expression of CD11b. Purified bone marrow neutrophils were treated with PBS or 100 pg/ml LPS overnight in the presence of GM-CSF, and harvested for scRNAseq analysis to examine their profiles of gene expression. Overall design: WT murine neutrophils were stimulated by GM-CSF, and then treated with either PBS or LPS overnight

本研究通过以超低剂量脂多糖(LPS)培养小鼠原代骨髓来源中性粒细胞，成功获得免疫增强型中性粒细胞。该类细胞优先表达CD74、CD44、CD86等共刺激分子，且CD11b的表达水平显著下调。我们将纯化后的骨髓中性粒细胞置于粒细胞-巨噬细胞集落刺激因子(GM-CSF)的培养体系中，分别以磷酸盐缓冲液(PBS)或100 pg/ml的LPS处理过夜，随后收集细胞进行单细胞RNA测序(scRNAseq)分析，以检测其基因表达谱。实验整体设计：野生型(WT)小鼠中性粒细胞先经GM-CSF刺激，随后分别用PBS或LPS处理过夜。