anti cd4 bv 421  (Thermo Fisher)

Journal: PLOS Pathogens

Article Title: Permissive lung neutrophils facilitate tuberculosis immunopathogenesis in male phagocyte NADPH oxidase-deficient mice

doi: 10.1371/journal.ppat.1012500

Figure Lengend Snippet: Pulmonary (A) CD11b + Ly6G + neutrophil, (B) CD90.2 + T cell, (C) CD19 + B220 + B cell, and (D) CD19 + B220 + GL7 + FAS + Germinal Center B cell populations of Mtb-infected mice at four weeks post-infection. The percentages of each immune cell among lung CD45 + cells and total cell counts are presented in bar graphs. (E) Pulmonary CFUs and neutrophil counts of female WT, male WT, female Nox2 -/- , and male Nox2 -/- mice were enumerated in a time-course dependent manner. Six-week old mice (n = 5 per group) were aerosol infected with Mtb K strain. The number of lung neutrophils were calculated at 0, 2, 3, and 4 weeks post-infection. Red bars and symbols represent statistical analysis between male Nox2 -/- mice and male WT mice. Black bars and symbols represent statistical analysis between male Nox2 -/- mice and female Nox2 -/- mice. Initial CFU = 300. The experiment was conducted once. The data are presented as the mean ± SD of five or six mice in each group. The significance of differences was determined, using the One-way ANOVA test. n . s ., not significant. ** p <0.01. (F) The compositions of the six major lung immune cell populations in Mtb-infected mice are presented in pie charts and t-SNE plots. The six major immune cells consist of neutrophils (red), macrophages (orange), alveolar macrophages (bright brown), dendritic cells (purple), B cells (blue), and T cells (green). The percentages of each immune cell among the sum of the six major immune cells are presented in pie charts.

Article Snippet: Brilliant violet (BV) 605-conjugated mAb against Thy1.2 and CD19; Allophycocyanin (APC)-conjugated mAb against CD45R (B220); BV 421-conjugated mAb against CD45; APC-R700-conjugated mAb against Siglec-F; Fluorescein isothiocyanate (FITC)-conjugated mAb against CD95 (FAS); BV 786-conjugated mAb against CXCR2; and V450- conjugated mAb against Ly6G were purchased from BD Bioscience (San Jose, CA, USA).

Techniques: Infection, Aerosol

Journal: PLOS Pathogens

Article Title: Permissive lung neutrophils facilitate tuberculosis immunopathogenesis in male phagocyte NADPH oxidase-deficient mice

doi: 10.1371/journal.ppat.1012500

Figure Lengend Snippet: Pulmonary (A) CD11b + Ly6G + neutrophil, (B) CD11b + Ly6G + CXCR2 lo CD62L lo immature neutrophil, CD11b + Ly6G + CXCR2 hi CD62L hi mature neutrophil, (C) CD19 + MHC-II + B cell, and ( D) CD90.2 + T cell populations of Mtb-infected mice at four weeks post-infection. The percentages of each immune cell among lung CD45 + cells or total neutrophils, and total cell counts are presented in bar graphs, along with flow cytometry plots. (E) The compositions of the six major lung immune cell populations in Mtb-infected mice are presented in pie charts and t-SNE plots. The six major immune cells consist of neutrophils (red), macrophages (orange), alveolar macrophages (bright brown), dendritic cells (purple), B cells (blue), and T cells (green). The percentages of each immune cell among the sum of the six major immune cells are presented in pie charts. The data are presented as the mean ± SD of four mice in each group. The significance of differences was determined, using the One-way ANOVA and Mann-Whitney U test. n . s ., not significant. * p < 0.05.

Article Snippet: Brilliant violet (BV) 605-conjugated mAb against Thy1.2 and CD19; Allophycocyanin (APC)-conjugated mAb against CD45R (B220); BV 421-conjugated mAb against CD45; APC-R700-conjugated mAb against Siglec-F; Fluorescein isothiocyanate (FITC)-conjugated mAb against CD95 (FAS); BV 786-conjugated mAb against CXCR2; and V450- conjugated mAb against Ly6G were purchased from BD Bioscience (San Jose, CA, USA).

Techniques: Infection, Flow Cytometry, MANN-WHITNEY

Journal: PLOS Pathogens

Article Title: Permissive lung neutrophils facilitate tuberculosis immunopathogenesis in male phagocyte NADPH oxidase-deficient mice

doi: 10.1371/journal.ppat.1012500

Figure Lengend Snippet: (A) Experimental design for in vivo administration of AM80 in Mtb infected mice. Six-week old male WT and Nox2 -/- mice (n = 5 per group) were aerosol infected with Mtb K strain. Starting from one week post-infection, 20 μg of AM80 was orally administered to each mouse three times a week (indicated by blue bars). At four weeks post-infection, all mice were autopsied, and immunological analysis, bacterial counting, and histopathological analysis were conducted (indicated by red arrow). Initial CFU = 320. (B) Mycobacterial CFUs in the lungs and spleens of each group at four weeks post-infection were analyzed by calculating the number of colonies and presented in bar graphs. (C) H&E staining was performed on the superior lobes of the right lung at four weeks post-infection to visualize the gross lung pathology. The inflamed area of the H&E-stained samples was quantified in terms of percentage and square millimeters and presented in bar graphs. (D) IFN-γ, IL-1α, IL-1β, IL-17A, IL-6, TNF-α, G-CSF, and BAFF levels in Mtb-infected mouse lung lysates were measured by ELISA and LEGENDplex. (E) IL-10, IL-4, IL-5, and TGF-β levels in Mtb-infected mouse lung lysates were measured by ELISA and LEGENDplex. The cytokine levels are presented in bar graphs. Pulmonary (F) CD11b + Ly6G + neutrophil, (G) CD11b + Ly6G + CXCR2 lo CD62L lo immature neutrophil, and CD11b + Ly6G + CXCR2 hi CD62L hi mature neutrophil populations of Mtb-infected mice at four weeks post-infection. The percentages of each immune cell among lung CD45 + cells or total neutrophils, and total cell counts are presented in bar graphs, along with flow cytometry plots. The experiment was conducted twice. The data are presented as the mean ± SD of five mice in each group. The significance of differences was determined, using the One-way ANOVA and Mann-Whitney U test. n . s ., not significant. * p < 0.05. ** p < 0.01.

Article Snippet: Brilliant violet (BV) 605-conjugated mAb against Thy1.2 and CD19; Allophycocyanin (APC)-conjugated mAb against CD45R (B220); BV 421-conjugated mAb against CD45; APC-R700-conjugated mAb against Siglec-F; Fluorescein isothiocyanate (FITC)-conjugated mAb against CD95 (FAS); BV 786-conjugated mAb against CXCR2; and V450- conjugated mAb against Ly6G were purchased from BD Bioscience (San Jose, CA, USA).

Techniques: In Vivo, Infection, Aerosol, Staining, Enzyme-linked Immunosorbent Assay, Flow Cytometry, MANN-WHITNEY

Journal: PLOS Pathogens

Article Title: Permissive lung neutrophils facilitate tuberculosis immunopathogenesis in male phagocyte NADPH oxidase-deficient mice

doi: 10.1371/journal.ppat.1012500

Figure Lengend Snippet: (A) Experimental design for in vivo neutralization of G-CSF in Mtb infected mice. Six-week old male WT and Nox2 -/- mice (n = 6 per group) were aerosol infected with Mtb K strain. Starting from one week post-infection, 30 μg of anti-G-CSF mAb was intraperitoneally administered to each mouse three times a week (indicated by blue bars). At four weeks post-infection, all mice were autopsied, and immunological analysis, bacterial counting, and histopathological analysis were conducted (indicated by red arrow). Initial CFU = 225. (B) Mycobacterial CFUs in the lungs and spleens of each group at four weeks post-infection were analyzed by calculating the number of colonies and presented in bar graphs. (C) H&E staining was performed on the superior lobes of the right lung at four weeks post-infection to visualize the gross lung pathology. The inflamed area of the H&E-stained samples was quantified in terms of percentage and square millimeters and presented in bar graphs. (D) IFN-γ, IL-1α, IL-1β, IL-17A, IL-6, TNF-α, G-CSF, and BAFF levels in Mtb-infected mouse lung lysates were measured by ELISA and LEGENDplex. (E) IL-10, IL-4, IL-5, and TGF-β levels in Mtb-infected mouse lung lysates were measured by ELISA and LEGENDplex. The cytokine levels are presented in bar graphs. Pulmonary (F) CD11b + Ly6G + neutrophil, (G) CD11b + Ly6G + CXCR2 lo CD62L lo immature neutrophil, and CD11b + Ly6G + CXCR2 hi CD62L hi mature neutrophil populations of Mtb-infected mice at four weeks post-infection. The percentages of each immune cell among lung CD45 + cells or total neutrophils, and total cell counts are presented in bar graphs, along with flow cytometry plots. The experiment was conducted three times. The data are presented as the mean ± SD of six mice in each group. The significance of differences was determined, using the One-way ANOVA test. n . s ., not significant. * p < 0.05. ** p < 0.01.

Article Snippet: Brilliant violet (BV) 605-conjugated mAb against Thy1.2 and CD19; Allophycocyanin (APC)-conjugated mAb against CD45R (B220); BV 421-conjugated mAb against CD45; APC-R700-conjugated mAb against Siglec-F; Fluorescein isothiocyanate (FITC)-conjugated mAb against CD95 (FAS); BV 786-conjugated mAb against CXCR2; and V450- conjugated mAb against Ly6G were purchased from BD Bioscience (San Jose, CA, USA).

Techniques: In Vivo, Neutralization, Infection, Aerosol, Staining, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Journal of visualized experiments : JoVE

Article Title: A Mouse Model for the Transition of Streptococcus pneumoniae from Colonizer to Pathogen upon Viral Co-Infection Recapitulates Age-Exacerbated Illness

doi: 10.3791/64419

Figure Lengend Snippet: Table of Materials

Article Snippet: CD103 BV 421 , BD Bioscience , BDB562771 , Clone: M290 DF 1:200.

Techniques: Flow Cytometry, Sterility, Modification, Staining, Blocking Assay, Ointment, Infection

Journal: Journal of visualized experiments : JoVE

Article Title: A Mouse Model for the Transition of Streptococcus pneumoniae from Colonizer to Pathogen upon Viral Co-Infection Recapitulates Age-Exacerbated Illness

doi: 10.3791/64419

Figure Lengend Snippet: Antibody panel 1.

Article Snippet: CD103 BV 421 , BD Bioscience , BDB562771 , Clone: M290 DF 1:200.

Techniques: Blocking Assay