Journal: Respiratory Research

Article Title: Targeting transitioning lung monocytes/macrophages as treatment strategies in lung disease related to environmental exposures

doi: 10.1186/s12931-024-02804-3

Figure Lengend Snippet: Inhalation of organic dust extract (ODE), lipopolysaccharide (LPS), and peptidoglycan (PGN) induce lung CCR2 + monocyte-macrophages (Mɸ) and monocytes. CCR2 RFP/+ mice were exposed once to ODE (25%), LPS (10 μg), PGN (100 μg), or saline control and euthanized at 48 h. Scatter plots with bars depict mean with SD delineating cells as total (gray), CCR2 + (green), and CCR2 − (red). A Total lung cells enumerated. B Representative contour plot of the four monocyte (mono)-Mɸ subpopulations across groups based upon CD11c and CD11b expression after removal of neutrophils gated from live CD45 + cells after excluding debris and doublets. C RFP ± staining by exposure group and subpopulation. D CD11c + CD11b lo alveolar (Alv) Mɸ, CD11c + CD11b hi activated (Act) Mɸ, CD11c int CD11b hi mono-Mɸ, and CD11c − CD11b. hi monocytes determined by multiplying lung cell % population by total lung cells enumerated from lung sample. Statistical analyses were performed with Kruskal–Wallis with Dunn’s test for multiple comparisons (# p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001) vs. respective saline. N = 19 (saline), 9 (ODE), 8 (LPS), 5 (PGN)

Article Snippet: Mouse tail snips were collected and shipped for DNA extraction and targeted CCR2 genotyping (TransnetYX, Cordova, TN) to confirm CCR2 KO.

Techniques: Saline, Control, Expressing, Staining

Journal: Respiratory Research

Article Title: Targeting transitioning lung monocytes/macrophages as treatment strategies in lung disease related to environmental exposures

doi: 10.1186/s12931-024-02804-3

Figure Lengend Snippet: Ly6C expression of monocyte-macrophage (Mɸ) and monocyte subpopulations following organic dust extract (ODE), lipopolysaccharide (LPS), and peptidoglycan (PGN) exposure. C57BL/6 mice were exposed once to ODE (25%), LPS (10 μg), PGN (100 μg), or saline control and euthanized at 48 h. Scatter plots with bars depict mean with SD delineating cells as CCR2 + (green) and CCR2- (red). Expression of Ly6C by percent ( A ) and mean fluorescence intensity (MFI) ( B ) across CD11cintCD11bhi monocyte-Mɸ and CD11c-CD11b + monocyte subpopulations as determined by flow cytometry. Statistical analyses were performed with Kruskal–Wallis with Dunn’s test for multiple comparisons (# p < 0.05 vs. respective saline) and (* p < 0.05 denoted by line with brackets denoting difference between same inhalant exposure by CCR2 RFP positive vs. negative). N = 15 (saline), 9 (ODE), 4 (LPS), 5 (PGN)

Article Snippet: Mouse tail snips were collected and shipped for DNA extraction and targeted CCR2 genotyping (TransnetYX, Cordova, TN) to confirm CCR2 KO.

Techniques: Expressing, Saline, Control, Fluorescence, Flow Cytometry

Journal: Respiratory Research

Article Title: Targeting transitioning lung monocytes/macrophages as treatment strategies in lung disease related to environmental exposures

doi: 10.1186/s12931-024-02804-3

Figure Lengend Snippet: Organic dust extract (ODE) and lipopolysaccharide (LPS) inhalation induce lung infiltration of CCR2 + NK and T cells. C57BL/6 mice were exposed once to ODE (25%), LPS (10 μg), PGN (100 μg), or saline control and euthanized at 48 h. Scatter plots with bars depict mean with SD delineating cells as total (gray), CCR2 + (green), and CCR2- (red). CD11c-Ly6G + neutrophils, CD19 + B cells, CD3-NK1.1 + NK cells, CD3 + CD4 + T cells, and CD3 + CD8 + T cell infiltrates determined by flow cytometry on live CD45 + cells after exclusion of debris and doublets with lung cell % populations multiplied by total lung cells enumerated from lung sample. Gating strategy depicted in Supplemental Fig. . Statistical analyses were performed with Kruskal–Wallis with Dunn’s test for multiple comparisons (# p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001) vs. respective saline. N = 19 (saline), 9 (ODE), 8 (LPS), 5 (PGN)

Article Snippet: Mouse tail snips were collected and shipped for DNA extraction and targeted CCR2 genotyping (TransnetYX, Cordova, TN) to confirm CCR2 KO.

Techniques: Saline, Control, Flow Cytometry

Journal: Respiratory Research

Article Title: Targeting transitioning lung monocytes/macrophages as treatment strategies in lung disease related to environmental exposures

doi: 10.1186/s12931-024-02804-3

Figure Lengend Snippet: Inhalant LPS-induced lung inflammatory responses are not reduced in CCR2 knock-out (KO) mice

Article Snippet: Mouse tail snips were collected and shipped for DNA extraction and targeted CCR2 genotyping (TransnetYX, Cordova, TN) to confirm CCR2 KO.

Techniques: Knock-Out

Journal: Respiratory Research

Article Title: Targeting transitioning lung monocytes/macrophages as treatment strategies in lung disease related to environmental exposures

doi: 10.1186/s12931-024-02804-3

Figure Lengend Snippet: Effects of systemic clodronate liposome delivery with LPS-induced lung inflammation, collagen, and infiltrating CCR2 + cells. Mice were pre-treated with vehicle (Veh) or clodronate (Clod) liposomes 24 h prior to a one-time treatment with LPS (10 μg) or saline (Sal) control and euthanized at 48 h. A Representative images from treatment groups stained by H&E, trichome, and CCR2 (red) with DAPI nuclei staining (blue) by confocal microscopy. Scatter plots with bars depict mean with SD of semi-quantitative lung inflammatory score ( B ) and integrated density of collagen ( C ), and CCR2 ( D ) quantified per each mouse. Statistical analyses were performed with Kruskal–Wallis with Dunn’s multiple comparison (inflammatory scores) and ANOVA with Tukey’s multiple comparison (collagen content and CCR2) (# p < 0.05 vs. respective saline) and (* p < 0.05 denoted by line with brackets denoting difference between groups). N = 8 (Veh + Sal), 8 (Clod + Sal), 8 (Veh + LPS), 9 (Clod + LPS). Line scale denotes 100 μm

Article Snippet: Mouse tail snips were collected and shipped for DNA extraction and targeted CCR2 genotyping (TransnetYX, Cordova, TN) to confirm CCR2 KO.

Techniques: Liposomes, Saline, Control, Staining, Confocal Microscopy, Comparison