Journal: PLOS One

Article Title: Glucocorticoids suppress early lung inflammation and impair control of SARS-CoV-2 in non-human primates

doi: 10.1371/journal.pone.0342849

Figure Lengend Snippet: A) Experimental design and sample collection. Created in part with BioRender under CC BY license, Nelson, C., (2025). B) Quantification of total lung lesions detected with 18 FDG-PET/CT imaging at the peak of the response, per animal and per group with mean. Animal #ID represented by color and shape in legend. C) Example 18 FDG-PET/CT images from control animal #DGT7 and GC treated animal #DHXD. D) Quantification of the lung lesion volume (dot size) and the metabolic intensity (normalized FDG uptake = SUV of lesion/SUV muscle). Significance calculated with 2way Anova multiple comparison test at the indicated timepoints between GC treatment and control. E) Quantification of subgenomic RNA of the SARS-CoV-2 N1 protein in copies per mL of BAL fluid. Individual animals and the mean of each group with standard error mean represented. Significance calculated with 2way Anova. Fold change between GC treatment and control indicated. Limit of detection (LOD) is 2,000 copies/mL fluid. F) Subgenomic N1 in copies per gram of tissue in the pulmonary lymph nodes, PET+ involved lung, and PET- uninvolved lung. Significance calculated with 2way Anova multiple comparison test. LOD is 1,000 copies per gram of tissue. p > 0.05 not shown, *p < 0.05, ***p < 0.001.

Article Snippet: Peptide pool consisted of Peptivator SARS-CoV-2 Prot_S Complete (Miltenyi Cat#130-127-953), Peptivator SARS-CoV-2 Prot_S B.1.617.2 Mutation (Miltenyi Cat#130-128-763), and Peptivator SARS-CoV-2 Prot_N (Miltenyi Cat# 130-126-699).

Techniques: Positron Emission Tomography-Computed Tomography, Imaging, Control, Comparison

Journal: PLOS One

Article Title: Glucocorticoids suppress early lung inflammation and impair control of SARS-CoV-2 in non-human primates

doi: 10.1371/journal.pone.0342849

Figure Lengend Snippet: A) Representative flow cytometry of SARS-CoV-2 specific CD4 and CD8 T cell responses (IFNγ + /TNF+) in blood at day 7 post-infection after ex vivo peptide stimulation with Spike and Nucleocapsid peptide pools, gated on live/CD45 + /CD3 + /CD95 + . B) Quantification of the frequency of SARS-CoV-2 specific CD4 and CD8 T cell responses in the blood overtime after subtracting background staining in unstimulated samples. Significance calculated with 2-way Anova and multiple comparison test. C) Representative flow cytometry of SARS-CoV-2 specific CD4 and CD8 T cell responses (IFNγ + /TNF+) in BAL at day 13 post-infection after ex vivo peptide stimulation with Spike and Nucleocapsid peptide pools, gated on live/CD45 + /CD3 + /CD95 + . D) Quantification of the frequency of SARS-CoV-2 specific CD4 and CD8 T cell responses in the BAL overtime after subtracting background staining in unstimulated samples. Significance calculated with 2-way Anova and multiple comparison test. E) Geometric mean fluorescence intensity (GMFI) of IFNγ production by IFNγ + SARS-CoV-2 specific CD4 and CD8 T cells. F) GMFI of TNF production by TNF+ SARS-CoV-2 specific CD4 and CD8 T cells. G) Frequency of IL-2+ of SARS-CoV-2 specific (IFNγ + /TNF+) CD4 and CD8 T cells in the BAL at day 13 post-infection. Significance calculated with unpaired t-test. H) Frequency of Granzyme B+ of SARS-CoV-2 specific (IFNγ + /TNF+) CD4 and CD8 T cells in the BAL at day 13 post-infection. I) Frequency of IL-17A+ of SARS-CoV-2 specific (IFNγ + /TNF+) CD4 and CD8 T cells in the BAL at day 13 post-infection. p > 0.05 ns or not shown, *p < 0.05, ***p < 0.001. ****p < 0.000.

Article Snippet: Peptide pool consisted of Peptivator SARS-CoV-2 Prot_S Complete (Miltenyi Cat#130-127-953), Peptivator SARS-CoV-2 Prot_S B.1.617.2 Mutation (Miltenyi Cat#130-128-763), and Peptivator SARS-CoV-2 Prot_N (Miltenyi Cat# 130-126-699).

Techniques: Flow Cytometry, Infection, Ex Vivo, Staining, Comparison, Fluorescence

Journal: PLOS One

Article Title: Glucocorticoids suppress early lung inflammation and impair control of SARS-CoV-2 in non-human primates

doi: 10.1371/journal.pone.0342849

Figure Lengend Snippet: A) Quantification of total B cells (CD3 - /CD20 + ) by flow cytometry in the blood over time. Mean of each group with standard error mean (SEM) represented. Significance calculated with 2way Anova. B) Representative flow cytometry plots and gating strategy for identifying total B cells (CD3 - /CD20 + ) from the BAL at day 4 after infection. C) Quantification of total B cells by flow cytometry in the BAL over time. Mean, SEM, and 2way Anova. D) Quantification of total B cells by scRNAseq (cluster 9) in the BAL over time. Mean, SEM, and 2way Anova. E) Differentially expressed genes in B cells (cluster 9) at day 4 post-infection between GC treatment vs. control. Red is upregulated with GC treatment, log 2 FC > 1 and adjusted p-value <0.05. Blue is downregulated with GC treatment, log 2 FC < −1 and adjusted p-value <0.05. Grey with large dot is adjusted p-value <0.05 but absolute |log 2 FC| < 1. Grey with small dot is adjusted p-value >0.05. F) Representative flow cytometry plots and gating strategy for identifying naïve B cells (CD3 - /CD20 + /IgD + ) and activated B cell (CD3 - /CD20 + /IgD - /CD95 + ) from the BAL at day 4 after infection. G) Quantification of naïve B cells by flow cytometry in the BAL over time. Mean, SEM, and 2way Anova. H) Quantification of activated B cells by flow cytometry in the BAL over time. Mean, SEM, and 2way Anova. I) Quantification of total B cells, J) naïve B cells, K) and activated B cells in the spleen, bone marrow, axillary lymph nodes, and pulmonary lymph nodes at necropsy day 13 post-infection. Significance calculated with 2way Anova. L) Representative flow cytometry plots of Spike-specific B cells (CD3 - /CD20 + /CD95 + /Spike-tetramer + ) isolated from the axillary or pulmonary lymph nodes at necropsy, day 13 post-infection. M) Quantification of Spike-specific B cells (CD3 - /CD20 + /CD95 + /Spike (B.1.617.2) tetramer + ) as a percentage of non-naïve (IgD - ) B cells in the spleen, bone marrow, axillary lymph nodes, and pulmonary lymph nodes at necropsy day 13 post-infection. N) Quantification of the frequency of IgG + expression by Spike-specific (tetramer + ) B cells in the pulmonary lymph node. Significance calculated with 2way Anova. p > 0.05 ns or not shown, *p < 0.05, ***p < 0.001. ****p < 0.0001.

Article Snippet: Peptide pool consisted of Peptivator SARS-CoV-2 Prot_S Complete (Miltenyi Cat#130-127-953), Peptivator SARS-CoV-2 Prot_S B.1.617.2 Mutation (Miltenyi Cat#130-128-763), and Peptivator SARS-CoV-2 Prot_N (Miltenyi Cat# 130-126-699).

Techniques: Flow Cytometry, Infection, Control, Isolation, Expressing