Journal: bioRxiv

Article Title: Caspase-4/11 exacerbates disease severity in SARS-CoV-2 infection by promoting inflammation and thrombosis

doi: 10.1101/2021.09.24.461743

Figure Lengend Snippet: a-b , WT and Casp11 -/- mice were infected with SARS-CoV-2 (MA10, 10 5 pfu). a , Lung sections from day 4 post-infection were stained with H&E to visualize lung damage and airway consolidation. b , Lung sections as in a were analyzed by the color deconvolution method to quantify cellularity as an indicator of cellular infiltration and alveolar wall thickening, ANOVA with Tukey’s multiple comparisons test. c,d , Lung homogenates from 2 or 4 days post-infection were analyzed by ELISA for detection of CXCL1, IL-1β, or IL-6, ANOVA with Tukey’s multiple comparisons test. e,f , Macrophages were purified from lungs of mice of the indicated genotype. The cells were infected with mouse adapted SARS-CoV-2 (MOI 1) for 24 h. Cell supernatants were analyzed by ELISA, or cellular RNA was analyzed by qRT-PCR for the indicated chemokine/cytokines, ANOVA with Tukey’s multiple comparisons test. g , Heatmap of significantly changed neutrophil-related genes comparing Casp11 -/- infected lungs versus WT(p<0.05). Expression scaling is relative to WT and Gsdmd -/- mice for comparisons. Numbers represent individual replicates and color indicates relative upregulation (red) or downregulation (blue) in gene expression. h , Lung sections of day 2 SARS-CoV-2-infected WT, Casp11 -/- and Gsdmd -/- mice (N=5) stained with neutrophil marker Ly6G and quantified in i. j , Flow cytometry of lungs homogenates from mice WT (N=4), Casp11 -/- (N=6), Gsdmd -/- (N=4) as in h and quantified in k . *p<0.05, **p<0.005.

Article Snippet: Cytokine/chemoking ELISAs were performed on lung homogenates or macrophage supernatants using R&D Systems Duoset ELISA kits (IL-6, DY406; IL-1b, DY401; CXCL1, DY453) according to the manufacturer’s instructions.

Techniques: Infection, Staining, Enzyme-linked Immunosorbent Assay, Purification, Quantitative RT-PCR, Expressing, Gene Expression, Marker, Flow Cytometry

Journal: Infection and Immunity

Article Title: The Cnes2 Locus on Mouse Chromosome 17 Regulates Host Defense against Cryptococcal Infection through Pleiotropic Effects on Host Immunity

doi: 10.1128/IAI.00697-15

Figure Lengend Snippet: B6.CBA-Cnes2 lungs have a heightened inflammatory response to C. neoformans infection characterized by increased expression of Th1 cytokines and chemokines. Whole lung protein was collected at 7 (A) or 14 (B to D) dpi with 104 CFU of C. neoformans 52D. ELISA was performed to determine the level of proinflammatory mediators (A), Th1/Th17-type cytokines (B), chemokines (C), and Th2-type cytokines (D). The expression of lung IL-4 mRNA was determined by quantitative RT-PCR. Data are shown as mean ± SEM (n = 4 to 6 mice per group) and are representative of two independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001 (using an unpaired Student t test).

Article Snippet: Contents of the following cytokines and chemokines in whole-lung protein samples were analyzed using DuoSet enzyme-linked immunosorbent assay (ELISA) kits (R&D Systems): TNF-α (DY410), IL-6 (DY406), IL-1β (DY401), CCL2/monocyte chemoattractant protein 1 (MCP-1) (DY479), IL-12/IL-23P40 (DY2398), IFN-γ (DY485), CXCL1/KC (DY453), CCL3/MIP-1α (DY450), CXCL2/MIP-2α (DY452), IL-4 (DY404), IL-5 (DY405), IL-13 (DY413), and IL-17A (DY421).

Techniques: Infection, Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Infection and Immunity

Article Title: The Cnes2 Locus on Mouse Chromosome 17 Regulates Host Defense against Cryptococcal Infection through Pleiotropic Effects on Host Immunity

doi: 10.1128/IAI.00697-15

Figure Lengend Snippet: Decreased Th2 type cytokine expression by CD4+ T cells from B6.CBA-Cnes2 lungs infected with C. neoformans. (A) Representative flow cytometry plots of lung lymphocytes from individual mice harvested at 21 dpi and restimulated with PMA-ionomycin, followed by intracellular staining for IFN-γ, IL-4, IL-5, and IL-13. Numbers shown are the percentage of cells in each gate relative to total cells in each plot and are expressed as mean ± SEM (n = 4 to 6 mice/group). (B) Frequency of CD4+ T cells expressing IFN-γ, IL-4, IL-5, and IL-13 per mouse at 21 dpi. Values represent means ± SEM (n = 4 to 6 mice). Data were pooled from two independent experiments. *, P < 0.05, using an unpaired Student t test.

Article Snippet: Contents of the following cytokines and chemokines in whole-lung protein samples were analyzed using DuoSet enzyme-linked immunosorbent assay (ELISA) kits (R&D Systems): TNF-α (DY410), IL-6 (DY406), IL-1β (DY401), CCL2/monocyte chemoattractant protein 1 (MCP-1) (DY479), IL-12/IL-23P40 (DY2398), IFN-γ (DY485), CXCL1/KC (DY453), CCL3/MIP-1α (DY450), CXCL2/MIP-2α (DY452), IL-4 (DY404), IL-5 (DY405), IL-13 (DY413), and IL-17A (DY421).

Techniques: Expressing, Infection, Flow Cytometry, Staining