Fig. 1: Intranasal β-glucan exposure generates environmentally adapted ApoE+CD11b+ AMs within the bronchoalveolar space.
a–e, Single-cell RNA sequencing (scRNA-seq) of the BALF of male 8- to 12-week-old C57BL/6J (WT) mice after intranasal stimulation with 200 µg β-glucan or PBS (n = 10,202 cells). Seven days after exposure, cells of three mice per condition were harvested and sorted for SSChi, Lin− (B220, CD19, CD3ε, Nk1.1, Ter-119), DRAQ7− singlets (n = 3 mice, one independent experiment). a, UMAP analysis of both conditions combined shows five different clusters. b, Heat map of the top ten highly expressed genes for each of the five clusters. c,d, UMAP from a separated by PBS (c; n = 4,845 cells) or β-glucan (d, n = 5,357 cells) condition. e, Percentage contribution of the five annotated clusters to overall cells split by conditions. f, 5-µm frozen section of the left lobe of the lung of a Ms4a3-creRosa26TOMATO mouse 7 days after β-glucan exposure stained with a 17-plex CODEX antibody panel. Overlaid images show the markers used to identify AM populations. Single stainings of these markers are shown in grayscale. Filled arrowheads indicate ApoE+CD11b+ AMs, whereas open arrowheads indicate CD11b− AMs. Scale bar, 100 µm (large image) and 10 µm (enlargements; n = one representative mouse of two independent experiments). g, Detection of Apoe mRNA expression in the AM subsets of the BALF 7 days after intranasal PBS or β-glucan exposure in WT mice by PrimeFlow (n = 3 mice pooled per group, one of two independent experiments shown). h,i, Flow cytometric quantification of absolute numbers (h) or frequency (i) of ApoE+CD11b+ AM (CD45+Siglec-F+CD64+CD11c+CD11b+) in the BALF in a time course from 1 to 21 days after β-glucan stimulation of WT mice (n = 9–10 mice, two independent experiments). Data in h and i are depicted as the mean ± s.d. Significance was assessed using ordinary one-way analysis of variance (ANOVA) with Tukey’s multiple comparisons.
