murine monoclonal anti il 17a antibody  (Bio X Cell)

Journal: NPJ Vaccines

Article Title: Vaccination with Mycoplasma pneumoniae membrane lipoproteins induces IL-17A driven neutrophilia that mediates Vaccine-Enhanced Disease

doi: 10.1038/s41541-022-00513-w

Figure Lengend Snippet: A Illustration of experimental timeline and outcome measures. B Lung lesion scores of vaccinated-then-challenged animals. Bronchoalveolar lavage fluid (BALF) concentrations of C TNF-α, D IL-1β, E IL-6, F IL-17A, and G KC in vaccinated-then-challenged animals. Positive correlations between disease severity (lung lesion scores) and BALF H IL-17A, and I KC concentrations. * p < 0.5, ** p < 0.1, *** p < 0.01, **** p < 0.001. Error bars for B indicate median and interquartile range and mean and SEM for C – G . Dotted lines for linear regression graphs indicate 95% confidence intervals. Each point represents data from an individual animal. Nonparametric lesion score data were analyzed via a one-way ANOVA on ranks (Kruskal–Wallis) with a Dunn’s post-hoc test for multiple pairwise comparisons. Parametric cytokine concentration data were analyzed via an ordinary one-way ANOVA with a Tukey’s post-hoc test for multiple pairwise comparisons. Linear regression was utilized to establish correlations.

Article Snippet: Starting 1 day prior to challenge (day −1) and continuing daily until the end of the study period (day 4), mice were intraperitoneally injected with 150 μg/250 μl/dose of either murine monoclonal anti-IL-17A antibody (BioXcell; clone 17F3, InVivoMAb anti-mouse IL-17A Cat#. BE0173) or the IgG1 isotype control antibody (BioXCell; clone MOPC-21, InVivoMAb IgG1 isotype control, Cat#. BE0083; San Antonio, TX).

Techniques: Concentration Assay

Journal: NPJ Vaccines

Article Title: Vaccination with Mycoplasma pneumoniae membrane lipoproteins induces IL-17A driven neutrophilia that mediates Vaccine-Enhanced Disease

doi: 10.1038/s41541-022-00513-w

Figure Lengend Snippet: Representative H&E stained lung sections ( A , C , E , G , I , K ) and RNAScope in situ hybridization processed slides staining IL-17A transcript (blue) and CD4 transcript (red) ( B , D , F , H , J , L ) from Sham-vaccinated/Mp-challenged animals (top), LAMPs-vaccinated/Mp-challenged animals (middle) and dLAMPs-vaccinated/Mp-challenged animals (bottom). Scale bars indicate 500 um (4x) or 100 um (10x).

Article Snippet: Starting 1 day prior to challenge (day −1) and continuing daily until the end of the study period (day 4), mice were intraperitoneally injected with 150 μg/250 μl/dose of either murine monoclonal anti-IL-17A antibody (BioXcell; clone 17F3, InVivoMAb anti-mouse IL-17A Cat#. BE0173) or the IgG1 isotype control antibody (BioXCell; clone MOPC-21, InVivoMAb IgG1 isotype control, Cat#. BE0083; San Antonio, TX).

Techniques: Staining, RNAscope, In Situ Hybridization

Journal: NPJ Vaccines

Article Title: Vaccination with Mycoplasma pneumoniae membrane lipoproteins induces IL-17A driven neutrophilia that mediates Vaccine-Enhanced Disease

doi: 10.1038/s41541-022-00513-w

Figure Lengend Snippet: Percentage ( A , E , I ) and number ( B , F , J ) of IL-17A positive cells when analyzing all live-single-cells (top), lymphocyte-like live-single cells (middle) and other-cells (bottom). Overlaid histograms ( C , G , K ) of cell count vs IL-17A signal on all live-single-cells (top), lymphocyte-like live-single cells (middle) and other-cells (bottom). Dot-plots ( D , H , L ) of Forward Scatter area vs IL-17A signal on all live-single-cells (top), lymphocyte-like live-single cells (middle) and other-cells (bottom). * p < 0.5, ** p < 0.1, *** p < 0.01, **** p < 0.001. Error bars indicate mean and SEM. Each point represents data from an individual animal. Data from single representative animals from each vaccination group are shown on histograms and dot-plots. Nonparametric percent frequency data were analyzed via a one-way ANOVA on ranks (Kruskal–Wallis) with a Dunn’s post-hoc test for multiple pairwise comparisons. Parametric cell count data were analyzed via an ordinary one-way ANOVA with a Tukey’s post-hoc test for multiple pairwise comparisons.

Article Snippet: Starting 1 day prior to challenge (day −1) and continuing daily until the end of the study period (day 4), mice were intraperitoneally injected with 150 μg/250 μl/dose of either murine monoclonal anti-IL-17A antibody (BioXcell; clone 17F3, InVivoMAb anti-mouse IL-17A Cat#. BE0173) or the IgG1 isotype control antibody (BioXCell; clone MOPC-21, InVivoMAb IgG1 isotype control, Cat#. BE0083; San Antonio, TX).

Techniques: Cell Counting

Journal: NPJ Vaccines

Article Title: Vaccination with Mycoplasma pneumoniae membrane lipoproteins induces IL-17A driven neutrophilia that mediates Vaccine-Enhanced Disease

doi: 10.1038/s41541-022-00513-w

Figure Lengend Snippet: H&E stained lung sections (left) and RNAScope in situ hybridization processed slides staining IL-17A transcript (blue) and CD4 transcript (red) (right) displaying vessels and airways to show that CD4 mRNA and IL-17A mRNA co-localization was more frequent in the areas of perivascular cuffing.

Article Snippet: Starting 1 day prior to challenge (day −1) and continuing daily until the end of the study period (day 4), mice were intraperitoneally injected with 150 μg/250 μl/dose of either murine monoclonal anti-IL-17A antibody (BioXcell; clone 17F3, InVivoMAb anti-mouse IL-17A Cat#. BE0173) or the IgG1 isotype control antibody (BioXCell; clone MOPC-21, InVivoMAb IgG1 isotype control, Cat#. BE0083; San Antonio, TX).

Techniques: Staining, RNAscope, In Situ Hybridization

Journal: NPJ Vaccines

Article Title: Vaccination with Mycoplasma pneumoniae membrane lipoproteins induces IL-17A driven neutrophilia that mediates Vaccine-Enhanced Disease

doi: 10.1038/s41541-022-00513-w

Figure Lengend Snippet: IL-17A producing CD3 + CD4 + cells as a percent of IL-17A positive lymphocyte-like cells ( A ) and raw counts per 50k analyzed events ( E ). IL-17A producing CD3 + CD4- cells as a percent of IL-17A positive lymphocyte-like cells ( B ) and raw counts per 50k analyzed events ( F ). IL-17A producing CD3-CD4 + cells as a percent of IL-17A positive lymphocyte-like cells ( C ) and raw counts per 50k analyzed events ( G ). IL-17A producing CD3-CD4- cells as a percent of IL-17A positive lymphocyte-like cells ( D ) and raw counts per 50k analyzed events ( E ). * p < 0.5, ** p < 0.1, *** p < 0.01, **** p < 0.001. Error bars indicate mean and SEM. Each point represents data from an individual animal. Nonparametric percent frequency data were analyzed via a one-way ANOVA on ranks (Kruskal–Wallis) with a Dunn’s post-hoc test for multiple pairwise comparisons. Parametric cell count data were analyzed via an ordinary one-way ANOVA with a Tukey’s post-hoc test for multiple pairwise comparisons.

Article Snippet: Starting 1 day prior to challenge (day −1) and continuing daily until the end of the study period (day 4), mice were intraperitoneally injected with 150 μg/250 μl/dose of either murine monoclonal anti-IL-17A antibody (BioXcell; clone 17F3, InVivoMAb anti-mouse IL-17A Cat#. BE0173) or the IgG1 isotype control antibody (BioXCell; clone MOPC-21, InVivoMAb IgG1 isotype control, Cat#. BE0083; San Antonio, TX).

Techniques: Cell Counting

Journal: NPJ Vaccines

Article Title: Vaccination with Mycoplasma pneumoniae membrane lipoproteins induces IL-17A driven neutrophilia that mediates Vaccine-Enhanced Disease

doi: 10.1038/s41541-022-00513-w

Figure Lengend Snippet: A Illustration of experimental timeline and outcome measures. B Numbers of lung-infiltrating leukocytes, proportion C and numbers D of lung-infiltrating neutrophils in vaccinated-then-challenged animals. E Positive correlations between lung-infiltrating neutrophil proportions and Lung Lesion Scores. F – I Correlations between proportions and numbers of lung-infiltrating neutrophils and IL-17A and KC concentrations. * p < 0.5, ** p < 0.1, *** p < 0.01, **** p < 0.001. Error bars for B – D indicate mean and SEM. Dotted lines for linear regression graphs indicate 95% confidence intervals. Each point represents data from an individual animal. Nonparametric percent frequency/proportion data were analyzed via a one-way ANOVA on ranks (Kruskal–Wallis) with a Dunn’s post-hoc test for multiple pairwise comparisons. Parametric cell count data were analyzed via an ordinary one-way ANOVA with a Tukey’s post-hoc test for multiple pairwise comparisons. Linear regression was utilized to establish correlations.

Article Snippet: Starting 1 day prior to challenge (day −1) and continuing daily until the end of the study period (day 4), mice were intraperitoneally injected with 150 μg/250 μl/dose of either murine monoclonal anti-IL-17A antibody (BioXcell; clone 17F3, InVivoMAb anti-mouse IL-17A Cat#. BE0173) or the IgG1 isotype control antibody (BioXCell; clone MOPC-21, InVivoMAb IgG1 isotype control, Cat#. BE0083; San Antonio, TX).

Techniques: Cell Counting

Journal: NPJ Vaccines

Article Title: Vaccination with Mycoplasma pneumoniae membrane lipoproteins induces IL-17A driven neutrophilia that mediates Vaccine-Enhanced Disease

doi: 10.1038/s41541-022-00513-w

Figure Lengend Snippet: A Illustration of experimental timeline and outcome measures. BALF concentrations of B IL-17A, C TNF-α, D IL-1β, E IL-6, and F KC in LAMPs-vaccinated/ Mp -challenged animals receiving an anti-IL-17A neutralizing monoclonal antibody (17F3) or isotype control (MOPC-21). BALF numbers of lung-infiltrating leukocytes ( G ), and proportion of H and number of ( I ) lung-infiltrating neutrophils. J Lung Lesion Scores and K bacterial loads of vaccinated-then-challenged animals treated with anti-IL-17A antibody or isotype control. * p < 0.5, ** p < 0.1, *** p < 0.01, **** p < 0.001. Error bars for J and K indicate median and interquartile range and mean and SEM for B – I . Each point represents data from an individual animal. Nonparametric lesion score, bacterial burden and percent frequency/proportion data were analyzed via an unpaired, two-tailed Mann–Whitney U -test. Parametric cytokine concentration and cell count data were analyzed via an unpaired, two-tailed t -test.

Article Snippet: Starting 1 day prior to challenge (day −1) and continuing daily until the end of the study period (day 4), mice were intraperitoneally injected with 150 μg/250 μl/dose of either murine monoclonal anti-IL-17A antibody (BioXcell; clone 17F3, InVivoMAb anti-mouse IL-17A Cat#. BE0173) or the IgG1 isotype control antibody (BioXCell; clone MOPC-21, InVivoMAb IgG1 isotype control, Cat#. BE0083; San Antonio, TX).

Techniques: Control, Two Tailed Test, MANN-WHITNEY, Concentration Assay, Cell Counting

Journal: NPJ Vaccines

Article Title: Vaccination with Mycoplasma pneumoniae membrane lipoproteins induces IL-17A driven neutrophilia that mediates Vaccine-Enhanced Disease

doi: 10.1038/s41541-022-00513-w

Figure Lengend Snippet: Anamnestic reactivation of IL-17A recall responses in LAMPs-vaccinated/ Mp -challenged animals results in the further production of TNF-α, IL-1β, IL-6, and KC. TNF-α and IL-1β can further induce the expression of the neutrophil chemotactic factor KC (Supplementary References , ), and in the presence of IL-6, further potentiate IL-17A production by helper T-cells (Supplementary Reference ), establishing a positive feedback loop of neutrophil recruitment and inflammation. Neutrophils also contribute to TNF-α production which can further potentiate KC production, contributing to the positive neutrophil recruitment loop that is associated with the more severe disease observed in Mp VED. ( Created in biorender.com by ABM ).

Article Snippet: Starting 1 day prior to challenge (day −1) and continuing daily until the end of the study period (day 4), mice were intraperitoneally injected with 150 μg/250 μl/dose of either murine monoclonal anti-IL-17A antibody (BioXcell; clone 17F3, InVivoMAb anti-mouse IL-17A Cat#. BE0173) or the IgG1 isotype control antibody (BioXCell; clone MOPC-21, InVivoMAb IgG1 isotype control, Cat#. BE0083; San Antonio, TX).

Techniques: Expressing

Journal: Journal of Neuroinflammation

Article Title: Interleukin-10 improves stroke outcome by controlling the detrimental Interleukin-17A response

doi: 10.1186/s12974-021-02316-7

Figure Lengend Snippet: IL-10 controls the IL-17A axis in the proinflammatory milieu of the postischemic brain. Relative gene expression of Il23a and Il1b in brain resident CD45 intermed /CD11b + microglia ( A ) and central nervous system-infiltrating CD45 high /CD11b + /CD11c − /MHCII − /Ly6g − /F4/80 + macrophages ( B ) purified 3, 7 and 14 days after tMCAO by fluorescence activated cell sorting from ischemic hemispheres. Expression levels were normalized to corresponding levels of splenic macrophages and microglia after sham operation. C Flow cytometric analysis of IL-17A produced by CD4 + and γδ T cells isolated from ischemic hemispheres of WT controls and Il10 −/− mice 7 days after tMCAO. D Flow cytometric analysis of number of infiltrating neutrophils in the ischemic hemispheres of WT controls and Il10 −/− mice 3 days after tMCAO. E Frequency of IL-17A producing γδ T cells purified from ischemic brains was analyzed 3 days following tMCAO in control mice (BSA) and mice receiving IL-10 intracerebral 3 h after tMCAO induction. A , B RT-qPCR gene expression data is represented as mean ± SEM of 4–7 WT, Flow cytometric data as mean ± SEM of 6 WT and 5 Il10 −/− ( C ), 9 WT and 7 Il10 −/− mice ( D ), and of 7 WT mice in each treatment group ( E ). Statistical significances were analyzed by one-way ANOVA with Bonferroni post hoc test ( A , B ) and by Student t test ( C – E ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Animals were treated intravenously with 500 μg of mouse monoclonal anti-murine IL-17A antibody (Clone MM17F3; 16.6 mg/kg of body weight) or with 500 μg of isotype-control antibody (IgG1, BioXCell, MOPC-21, BE0083) 3 h and 3 days after reperfusion.

Techniques: Gene Expression, Purification, Fluorescence, FACS, Expressing, Produced, Isolation, Control, Quantitative RT-PCR

Journal: Journal of Neuroinflammation

Article Title: Interleukin-10 improves stroke outcome by controlling the detrimental Interleukin-17A response

doi: 10.1186/s12974-021-02316-7

Figure Lengend Snippet: Neutralization of IL-17A abolishes the worse outcome of Il10 −/− mice. Triphenyltetrazolium chloride staining was used for evaluation of infarct volume at day 7 of IL-17A antibody ( A ) or IgG control ( C ) treated Il10 −/− and WT control mice. Bederson score was performed 7 days after tMCAO IL-17A antibody ( B ) or in IgG control ( D ) treated group. Infarct data are presented as mean ± SEM of 9–11 WT and 6–9 Il10 −/− mice per group. Bederson score as mean ± SEM of 9–11 WT and 7–10 Il10 −/− mice per group. Statistical significances were analyzed by Student t test ( A , C ) and Mann–Whitney U test ( B , D ). * P < 0.05

Article Snippet: Animals were treated intravenously with 500 μg of mouse monoclonal anti-murine IL-17A antibody (Clone MM17F3; 16.6 mg/kg of body weight) or with 500 μg of isotype-control antibody (IgG1, BioXCell, MOPC-21, BE0083) 3 h and 3 days after reperfusion.

Techniques: Neutralization, Staining, Control, MANN-WHITNEY

Journal: Journal of Neuroinflammation

Article Title: Interleukin-10 improves stroke outcome by controlling the detrimental Interleukin-17A response

doi: 10.1186/s12974-021-02316-7

Figure Lengend Snippet: T cell-specific blockade of IL-10R leads to increased frequencies of IL-17A + CD4 + and γδ T cells. Flow cytometric analysis of IL-17A produced by CD4 + ( A ) and γδ ( B ) T cells isolated from ischemic hemispheres of CD4 Cre Il10Ra wt/wt controls and CD4 Cre Il10Ra fl/fl 7 days after tMCAO. C Flow cytometric analysis of IL-10R (CD210) on IL-17A + and IL-17A − γδ T cells isolated from ischemic hemispheres of C57BL/6 mice 7 days after tMCAO. D Flow cytometric analysis of IL-17A produced by γδ T cells isolated from ischemic hemispheres of TCR δ CreER Il10Ra wt/wt controls and TCR δ CreER Il10Ra fl/fl 7 days after tMCAO. Flow cytometric data is represented as mean ± SEM of 9 CD4 Cre Il10Ra wt/wt and 8 CD4 Cre Il10Ra fl/fl ( A , B ), 6 WT ( C ) and 6 TCR δ CreER Il10Ra wt/wt and 7 TCR δ CreER Il10Ra fl/fl ( D ) mice per group. Statistical significances were analyzed by Student t test ( A – D ). * P < 0.05, ** P < 0.01. MFI mean fluorescent intensity

Article Snippet: Animals were treated intravenously with 500 μg of mouse monoclonal anti-murine IL-17A antibody (Clone MM17F3; 16.6 mg/kg of body weight) or with 500 μg of isotype-control antibody (IgG1, BioXCell, MOPC-21, BE0083) 3 h and 3 days after reperfusion.

Techniques: Produced, Isolation

Journal: Journal of Neuroinflammation

Article Title: Interleukin-10 improves stroke outcome by controlling the detrimental Interleukin-17A response

doi: 10.1186/s12974-021-02316-7

Figure Lengend Snippet: Control of IL-17A production in γδ but not CD4 + T cells is dependent on IL-10R expression on Foxp3 + Tregs. A Flow cytometric analysis of the IL-10R (CD210) Mean Fluorescent Intensity (MFI) on Foxp3 + and Foxp3 − CD4 + T cells in FIR-tiger mice. Flow cytometric analysis of IL-17A produced by CD4 + ( B ) and γδ ( C ) T cells isolated from ischemic hemispheres of Foxp3 Cre Il10Ra wt/wt and Foxp3 Cre Il10Ra fl/fl controls 7 days after tMCAO. ( D) Proposed mechanism of protection from ischemic brain injury induced by IL-10. Flow cytometric data is presented as mean ± SEM of 6 FIR-tiger ( A ) and 6 Foxp3 Cre Il10Ra wt/wt and 7 Foxp3 Cre Il10Ra fl/fl mice per group ( B , C ). Statistical significances were analyzed by Student t test ( A – C ). ** P < 0.01 and *** P < 0.01

Article Snippet: Animals were treated intravenously with 500 μg of mouse monoclonal anti-murine IL-17A antibody (Clone MM17F3; 16.6 mg/kg of body weight) or with 500 μg of isotype-control antibody (IgG1, BioXCell, MOPC-21, BE0083) 3 h and 3 days after reperfusion.

Techniques: Control, Expressing, Produced, Isolation