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. 2021 Sep 20;10:e69951. doi: 10.7554/eLife.69951

Figure 5. Trained immunity of alveolar macrophages (AMs) mediates rapid protection induced by vaccination.

(A) Heatmap of differentially expressed genes (DEGs) in lungs of inactivated whole cell (IWC) (LAC-4)-immunized and control Rag1-/- mice at day 7 after immunization. (B) Top 10 Gene Ontology (GO) terms of upregulated DEGs in IWC-immunized Rag1-/- mice at day 7. (C) In vitro model of IWC (LAC-4)-trained AMs. (D) C57BL/6 mice were immunized with IWC (LAC-4), and recall responses of trained AMs to IWC (LAC-4) were evaluated ex vivo by detection of TNF-α production at 2 hr after stimulation. For (C) and (D), data are mean ± SD. n = 3. *p<0.05, **p<0.01, two-tailed unpaired t test. (E) Schema of evaluating roles of AMs in BALF of PBS or IWC (LAC-4)-immunized C57BL/6 mice. Bacterial burdens of lung and clinical scores at 24 hr post infection (hpi) were measured (n = 5–9). Each dot means a mice, and the line represents the median. *p<0.05, **p<0.01 determined by Mann–Whitney U test. (F) WT mice were immunized with IWC (LAC-4) for 7 days. Mice were treated intraperitoneally with anti-TNF-α antibody or isotype control then were challenged with lethal LAC-4 1 hr later. The survival of mice was monitored (n = 5). *p<0.05, calculated by log-rank test. Data are representative of two independent experiments.

Figure 5—source data 1. Raw data for Figure 5.

Figure 5.

Figure 5—figure supplement 1. Transcriptional difference at day 7 after immunization of inactivated whole cell (IWC) (LAC-4) in Rag1-/- mice.

Figure 5—figure supplement 1.

(A) Numbers of upregulated and downregulated differentially expressed genes (DEGs) of lungs in IWC-7d and control mice. (B) Top 10 mouse gene atlas terms of upregulated genes in lungs of IWC-7d vs. control mice analyzed by Enrichr. (C) The purity of alveolar macrophages (AMs) after magnetic-activated cell sorting (MACS) sorting is above 95%.