anti il 17 pe cy7  (Miltenyi Biotec)

Journal: eLife

Article Title: Short-range interactions between fibrocytes and CD8 + T cells in COPD bronchial inflammatory response

doi: 10.7554/eLife.85875

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-IL-17-PE-Cy7 (mouse monoclonal) , Miltenyi Biotec , Cat. #:130-120-413, RRID: AB_2752086 , FC (1:20).

Techniques: Blocking Assay, Recombinant

Journal: bioRxiv

Article Title: Prime-pull immunization of mice with a BcfA-adjuvanted vaccine elicits mucosal immunity and prevents SARS CoV-2 infection and pathology

doi: 10.1101/2022.04.06.487394

Figure Lengend Snippet: (A). Prime-pull immunization strategy to elicit mucosal immune response in C57/BL mice. Mice (6/group) were injected i.m. on d0 and inoculated i.n. on d28 with various vaccine formulations, with analysis of T cell populations in the lungs conducted on d42 (2 weeks post-boost). Mice were injected i.v. with CD45-PE antibody 10 min before euthanasia to distinguish between resident (CD45-) and circulating (CD45+) T cells. (B) The percentage and number of CD45-CD3+CD4+CD44+CD62L-CD69+ antigen-experienced CD4+ TRM, and (C) the percentage and number of IFNγ+ (D), IL-5+ (E) and IL-17+ cells is shown. Gating strategy in . (F) The percentage and number of CD45-CD3+CD8+CD44+CD62L-CD69+ antigen-experienced CD8+ TRM and (G) IFNγ+ subset is shown. One-way ANOVA with Tukey’s multiple comparisons was used to detect differences between all experimental groups. Significance is indicated above for each group (∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001). The data are representative of 2 independent experiments. Mean and SEM are displayed.

Article Snippet: Following permeabilization (eBioscience Cat. 00-8333-56), intracellular staining (30 minutes at 4°C) was done using a cocktail of the following antibodies: IFNγ FITC (clone XMG1.2) (eBioscience Cat. 11-7311-82), IL-17 PE CY7 (clone eBio17B7) (eBioscience Cat. 25-7177-82), IL-5 APC (clone TRFK5) (BD Cat. 554396).

Techniques: Injection

Journal: bioRxiv

Article Title: Prime-pull immunization of mice with a BcfA-adjuvanted vaccine elicits mucosal immunity and prevents SARS CoV-2 infection and pathology

doi: 10.1101/2022.04.06.487394

Figure Lengend Snippet: Mice were injected with CD45 PE antibody 10 minutes before sac to distinguish between resident (CD45-) and circulating (CD45+) T cells. PMA/Ionomycin stimulated cells stained with surface markers to identify antigen experienced CD4+CD44+CD62L-CD69+ CD4+ cells were fixed, pearmeabilized and followed by intracellular cytokine staining (ICS) to analyze the production of IFNγ, IL-5 and IL-17. Shown is the sequential gating strategy to identify lymphocytes by forward and side scatter, single cells, live cells, CD45-cells, CD3+CD4+ cells, CD44+CD62L-cells and CD69+ cells. The proportion of IFNγ+, IL-17+ and IL-5+ cells within the CD3+CD4+CD45-CD44+CD62L-CD69+ fraction was determined. The same sequential gating strategy was used to identify CD45+ cells.

Article Snippet: Following permeabilization (eBioscience Cat. 00-8333-56), intracellular staining (30 minutes at 4°C) was done using a cocktail of the following antibodies: IFNγ FITC (clone XMG1.2) (eBioscience Cat. 11-7311-82), IL-17 PE CY7 (clone eBio17B7) (eBioscience Cat. 25-7177-82), IL-5 APC (clone TRFK5) (BD Cat. 554396).

Techniques: Injection, Staining

Journal: bioRxiv

Article Title: Prime-pull immunization of mice with a BcfA-adjuvanted vaccine elicits mucosal immunity and prevents SARS CoV-2 infection and pathology

doi: 10.1101/2022.04.06.487394

Figure Lengend Snippet: Lung cells (from mice in (6/group) were analyzed for the presence of CD45+ circulating T cells. (A) The percentage and number of CD45+CD3+CD4+CD44+CD62L-CD69+ antigen-experienced CD4+ T RM . (B) The percentage and number of IFNγ+ (C) IL-5+ and (D) IL-17+ cells is shown. (E) The percentage and of CD45+CD3+CD8+CD44+CD62L-CD69+ antigen-experienced CD8+ T RM. (F) The percentage and number of IFNγ+ subset is shown. (G) Splenocytes were isolated and processed into single cell suspensions before stimulation with 1μg S protein. IL-17 and IL-5 present in the culture supernatant 3 days post-stimulation was determined by ELISA. Differences between all experimental groups were analyzed by one-way ANOVA with Tukey’s multiple comparisons. Significance is indicated above for each group (∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001). Mean and SEM are shown.

Article Snippet: Following permeabilization (eBioscience Cat. 00-8333-56), intracellular staining (30 minutes at 4°C) was done using a cocktail of the following antibodies: IFNγ FITC (clone XMG1.2) (eBioscience Cat. 11-7311-82), IL-17 PE CY7 (clone eBio17B7) (eBioscience Cat. 25-7177-82), IL-5 APC (clone TRFK5) (BD Cat. 554396).

Techniques: Isolation, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Prime-pull immunization of mice with a BcfA-adjuvanted vaccine elicits mucosal immunity and prevents SARS CoV-2 infection and pathology

doi: 10.1101/2022.04.06.487394

Figure Lengend Snippet: Lung cells were stimulated for 6 hours with S-derived peptide pools (S1 and S2) at a concentration of 1μg/ml in the presence of protein transport inhibitors. Surface staining and ICS was conducted to analyze both CD45-resident and CD45+ circulating T cells. (A) The percentage and number of CD45-CD3+CD4+CD44+CD62L-CD69+ antigen-experienced CD4+ T RM . (B) The percentage and number of IFNγ+ (C) IL-5+ and (D) IL-17+ cells is shown. (E) The percentage and of CD45+CD3+CD4+CD44+CD62L-CD69+ antigen-experienced CD4+ T RM. (F) The percentage and number of IFNγ+ (G) IL-5+ and (H) IL-17+ subsets is shown.. One-way ANOVA with Tukey’s multiple comparisons was used to detect differences between all experimental groups. Significance is indicated above for each group (∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001). N= 5 animals per group, mean and SEM of the results are displayed.

Article Snippet: Following permeabilization (eBioscience Cat. 00-8333-56), intracellular staining (30 minutes at 4°C) was done using a cocktail of the following antibodies: IFNγ FITC (clone XMG1.2) (eBioscience Cat. 11-7311-82), IL-17 PE CY7 (clone eBio17B7) (eBioscience Cat. 25-7177-82), IL-5 APC (clone TRFK5) (BD Cat. 554396).

Techniques: Derivative Assay, Concentration Assay, Staining

Journal: bioRxiv

Article Title: Prime-pull immunization of mice with a BcfA-adjuvanted vaccine elicits mucosal immunity and prevents SARS CoV-2 infection and pathology

doi: 10.1101/2022.04.06.487394

Figure Lengend Snippet: (A) Prime-pull immunization strategy to elicit mucosal immune response in C57/BL or IL-17 KO mice. At 2 weeks post-boost mice (10 /group) were challenged with the mouse adapted SARS CoV2 strain (MA10). The percent of original body weight on each day post-challenge was calculated through day 10 post infection (5 mice/group) in (B) C57BL/6 mice and (C) IL-17 KO mice. Two-way ANOVA with Tukey’s multiple comparisons was used to detect differences between all experimental groups in B and Student’s t-test in C. Viral titer expressed as log Median Tissue Culture Infectious Dose (TCID50) was quantified from nasal septum (D) and lung homogenates (E) of C57BL/6 mice and nasal septum (F) and lungs (G) of IL-17 KO mice obtained on day 2 post infection (4-5 mice/group). One-way ANOVA with Tukey’s multiple comparisons was used to detect differences between experimental groups in D and E, and Student’s t-test in F and G. Mean and SEM of the results are displayed. Significance is indicated above for each group (∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001).

Article Snippet: Following permeabilization (eBioscience Cat. 00-8333-56), intracellular staining (30 minutes at 4°C) was done using a cocktail of the following antibodies: IFNγ FITC (clone XMG1.2) (eBioscience Cat. 11-7311-82), IL-17 PE CY7 (clone eBio17B7) (eBioscience Cat. 25-7177-82), IL-5 APC (clone TRFK5) (BD Cat. 554396).

Techniques: Infection

Journal: bioRxiv

Article Title: Prime-pull immunization of mice with a BcfA-adjuvanted vaccine elicits mucosal immunity and prevents SARS CoV-2 infection and pathology

doi: 10.1101/2022.04.06.487394

Figure Lengend Snippet: Formalin fixed, paraffin embedded lungs harvested on d3 post-infection were sectioned and stained with H&E to evaluate inflammation and tissue damage. (A) Unimmunized IL-17 KO mice, (B) S/A/B_S/B immunized mice. Semi-quantitative scoring of (C) alveolar wall thickness (D) presence of alveolar macrophages (E) degeneration and necrosis and (F) presence of lymphocytes and plasma cells is shown 4-5 samples/group, evaluated by Student’s t-test. *, p< 0.05, ***, p<0.001.

Article Snippet: Following permeabilization (eBioscience Cat. 00-8333-56), intracellular staining (30 minutes at 4°C) was done using a cocktail of the following antibodies: IFNγ FITC (clone XMG1.2) (eBioscience Cat. 11-7311-82), IL-17 PE CY7 (clone eBio17B7) (eBioscience Cat. 25-7177-82), IL-5 APC (clone TRFK5) (BD Cat. 554396).

Techniques: Formalin-fixed Paraffin-Embedded, Infection, Staining

Journal: Frontiers in Immunology

Article Title: XBP-1s Promotes B Cell Pathogenicity in Chronic GVHD by Restraining the Activity of Regulated IRE-1α-Dependent Decay

doi: 10.3389/fimmu.2021.705484

Figure Lengend Snippet: Effects of XBP-1s and IRE-1α on B cell ability to stimulate allogeneic T cells in vitro . B cells isolated from WT, XBP-1 KO , IRE-1α KO , and DKO on B6 background mice are stimulated with 1 μg/ml LPS and 10 ng/ml IL-4 for 24 hrs. T cells were isolated from FVB mice and labeled with CFSE and then incubated with activated B cells for another 4 days. The representative flow panels of CFSE dilution and percentages of CFSE diluted cells on CD4 (A, B) and CD8 (C, D) T cells. T cells were stimulated with PMA and Ionomycin for 4 h before intracellular cytokine staining for cytokine detection. The representative flow panels and the percentages of IFN-γ (E–G) , TNF-α (H–J) or IL-17 (K–M) among proliferated (CFSE low ) CD4 and CD8 T cells were shown. Data show representative flow cytometry plots and graphs from three independently repeated experiments. Statistics were performed using ordinary one-way ANOVA with Tukey’s multiple comparison test. *p < 0.05, **p < 0.005, ***p < 0.0005, and ****p < 0.0001 when compared to WT. # p < 0.05, ## p < 0.005, ### p < 0.0005, and #### p < 0.0001 when compared to XBP-1 KO group.

Article Snippet: Antibodies were purchased as followed: Anti-CD4-V450, anti-CD8α-APCcy7, anti-B220-V450, anti-CD138-Pe-cy7, anti-CD86-Pe-cy5, anti-FAS-PE, anti-GL-7-APC, anti-CD40-APC, anti-MHCII-FITC, anti-IL-4-PE, anti-IL-5-PE, anti-IgM-Pe-cy7, anti-IgG1-APC, anti-IFN-r-Percp5.5, anti-TNF-α-PE, and anti-IL-17-Pe-cy7 were purchased from BD Biosciences (Franklin Lakes, NJ).

Techniques: In Vitro, Isolation, Labeling, Incubation, Staining, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: XBP-1s Promotes B Cell Pathogenicity in Chronic GVHD by Restraining the Activity of Regulated IRE-1α-Dependent Decay

doi: 10.3389/fimmu.2021.705484

Figure Lengend Snippet: Effect of XBP-1s and IRE-1α on B cell ability to stimulate allogeneic T cells during cGVHD. Lethally irradiated BALB/c mice were transplanted with 5 x 10 6 TCD-BM cells from WT, XBP-1 KO , IRE-1α KO , and DKO mice on a B6 background with 0.35 - 0.5 x 10 6 splenocytes (n=20). Recipient mice were euthanized on day 28 after BMT and single cells were isolated from recipient spleens. The expression of Foxp3 transcription factor (A, B) was measured by flow cytometry analysis. The cytokine levels in CD4 and CD8 T cells including IL-4/IL-5 (C, D) , IFN-γ (E–G) , and IL-17 (H–J) were determined after intracellular staining by flow cytometry analysis. Data show the representative result from three independently repeated experiments. 20 recipient mice were used for each experiment. Statistics were performed using two-way ANOVA with Tukey’s multiple comparison test. *p < 0.05, **p < 0.005, ***p < 0.0005, and ****p < 0.0001 when compared to WT. # p < 0.05, ## p < 0.005, ### p < 0.0005, and #### p < 0.0001 when compared to XBP-1 KO group.

Article Snippet: Antibodies were purchased as followed: Anti-CD4-V450, anti-CD8α-APCcy7, anti-B220-V450, anti-CD138-Pe-cy7, anti-CD86-Pe-cy5, anti-FAS-PE, anti-GL-7-APC, anti-CD40-APC, anti-MHCII-FITC, anti-IL-4-PE, anti-IL-5-PE, anti-IgM-Pe-cy7, anti-IgG1-APC, anti-IFN-r-Percp5.5, anti-TNF-α-PE, and anti-IL-17-Pe-cy7 were purchased from BD Biosciences (Franklin Lakes, NJ).

Techniques: Irradiation, Isolation, Expressing, Flow Cytometry, Staining

Journal: International Journal of Molecular Sciences

Article Title: Extensive Histopathological Characterization of Inflamed Bowel in the Dextran Sulfate Sodium Mouse Model with Emphasis on Clinically Relevant Biomarkers and Targets for Drug Development

doi: 10.3390/ijms22042028

Figure Lengend Snippet: List of antibodies used in the study.

Article Snippet: anti-IL-17A , Rat monoclonal, clone TC11-18H10; Novus Biologicals, Littleton, CO, USA , 1:50 , EDTA–Citrate pH 7.8.

Techniques: