Journal: BMC Medicine

Article Title: Remission of autoimmune diabetes by anti-TCR combination therapies with anti-IL-17A or/and anti-IL-6 in the IDDM rat model of type 1 diabetes

doi: 10.1186/s12916-020-1503-6

Figure Lengend Snippet: Effects of anti-TCR combination therapies on the metabolic profile of IDDM rats after diabetes manifestation . a–d Blood glucose concentration (mmol/l) changes are shown for the responding rats in response to the different anti-TCR combination therapies a with anti-IL-6 (5/7), b anti-IL-17A (7/10) or c in the triple combination (6/10) compared to d the normoglycaemic healthy ( n = 6) and to the acutely diabetic untreated IDDM rats ( n = 6). The first dashed line at day 0 indicates the start of therapy (first biopsy) and the second dashed line at day 5 indicates the end of therapy (second biopsy). e Serum C-peptide concentration changes (pmol/l) are shown for rats responding and non-responding to the different combination therapies of anti-TCR with anti-IL-6 or anti-IL-17A alone or in combination. Data are mean values ± SEM. Comparison of the different experimental groups by one-way ANOVA followed by Bonferroni test *** p < 0.001 to the healthy control, $ p < 0.05 to anti-TCR combination with anti-IL-6, §§§ p < 0.001 to anti-TCR combination with anti-IL-17A, and ### p < 0.001 to triple combination for each observation time point

Article Snippet: Group 1 ( n = 6) comprised healthy, normoglycaemic IDDM rats which received 0.9% NaCl solution only, group 2 ( n = 6) comprised acutely diabetic IDDM rats which received IgG (I5381; Merck, Darmstadt, Germany; diluted with 0.9% NaCl solution) and groups 3, 4 and 5 ( n = 4 each) comprised acutely diabetic IDDM rats treated for 5 consecutive days in a monotherapy either with a rat-specific anti-TCR antibody (Clone: R73; Bio-Rad, Munich, Germany) (0.5 mg/kg b.wt. i.v.), rat-specific anti-IL-6 (0.01 mg/kg b.wt. i.v.) (Clone: MAB 561 , R&D Systems, Wiesbaden-Nordenstadt, Germany) or with a murine specific anti-IL-17A (0.1 mg/kg b.wt. i.v . ) (Clone: 17F3, eBioscience, Frankfurt, Germany ) .

Techniques: Concentration Assay

Journal: BMC Medicine

Article Title: Remission of autoimmune diabetes by anti-TCR combination therapies with anti-IL-17A or/and anti-IL-6 in the IDDM rat model of type 1 diabetes

doi: 10.1186/s12916-020-1503-6

Figure Lengend Snippet: Relation between initial blood glucose concentration and β cell mass after end of therapy. a After anti-TCR combination therapy with anti-IL-6 or b with anti-IL-17A or c with both cytokine antibodies together. The β cell mass showed the highest values after triple combination, followed by the double combination with anti-IL-17A and the lowest values after double combination with anti-IL-6. Remarkably, the starting blood glucose concentrations granting therapy success differed between the three analysed groups without a blood glucose concentration window for anti-TCR combination with anti-IL-6 and with starting blood glucose concentrations below 13 mmol/l for the anti-TCR combination with anti-IL-17A and 17 mmol/l for the triple combination compared to those above these glucose values

Article Snippet: Group 1 ( n = 6) comprised healthy, normoglycaemic IDDM rats which received 0.9% NaCl solution only, group 2 ( n = 6) comprised acutely diabetic IDDM rats which received IgG (I5381; Merck, Darmstadt, Germany; diluted with 0.9% NaCl solution) and groups 3, 4 and 5 ( n = 4 each) comprised acutely diabetic IDDM rats treated for 5 consecutive days in a monotherapy either with a rat-specific anti-TCR antibody (Clone: R73; Bio-Rad, Munich, Germany) (0.5 mg/kg b.wt. i.v.), rat-specific anti-IL-6 (0.01 mg/kg b.wt. i.v.) (Clone: MAB 561 , R&D Systems, Wiesbaden-Nordenstadt, Germany) or with a murine specific anti-IL-17A (0.1 mg/kg b.wt. i.v . ) (Clone: 17F3, eBioscience, Frankfurt, Germany ) .

Techniques: Concentration Assay

Journal: BMC Medicine

Article Title: Remission of autoimmune diabetes by anti-TCR combination therapies with anti-IL-17A or/and anti-IL-6 in the IDDM rat model of type 1 diabetes

doi: 10.1186/s12916-020-1503-6

Figure Lengend Snippet: Morphometric analyses of β cells and immune cells in IDDM rats after anti-TCR combination therapies. Changes in the rate of a proliferation, b apoptosis, c islet infiltration score and d pancreatic β cell mass after successful anti-TCR combination therapy with anti-IL-6 or anti-IL-17A alone or combined together after diabetes manifestation. Measurements were performed immediately before therapy (first biopsy), at the end of therapy (second biopsy) and 60 days after the end of therapy Data are mean values ± SEM. Comparison of the different experimental groups by one-way ANOVA followed by Bonferroni test *** p < 0.001, ** p < 0.01 and * p < 0.05 to the healthy control, $$$ p < 0.001 to anti-TCR combination with anti-IL-6, §§§ p < 0.001 and §§ p < 0.01 to anti-TCR combination with anti-IL-17A, ### p < 0.001 and ## p < 0.01 to triple combination for each observation time point. Numbers of pancreases analysed as given in Fig.

Article Snippet: Group 1 ( n = 6) comprised healthy, normoglycaemic IDDM rats which received 0.9% NaCl solution only, group 2 ( n = 6) comprised acutely diabetic IDDM rats which received IgG (I5381; Merck, Darmstadt, Germany; diluted with 0.9% NaCl solution) and groups 3, 4 and 5 ( n = 4 each) comprised acutely diabetic IDDM rats treated for 5 consecutive days in a monotherapy either with a rat-specific anti-TCR antibody (Clone: R73; Bio-Rad, Munich, Germany) (0.5 mg/kg b.wt. i.v.), rat-specific anti-IL-6 (0.01 mg/kg b.wt. i.v.) (Clone: MAB 561 , R&D Systems, Wiesbaden-Nordenstadt, Germany) or with a murine specific anti-IL-17A (0.1 mg/kg b.wt. i.v . ) (Clone: 17F3, eBioscience, Frankfurt, Germany ) .

Techniques:

Journal: BMC Medicine

Article Title: Remission of autoimmune diabetes by anti-TCR combination therapies with anti-IL-17A or/and anti-IL-6 in the IDDM rat model of type 1 diabetes

doi: 10.1186/s12916-020-1503-6

Figure Lengend Snippet: Immune cell infiltration in pancreatic islets of IDDM rats after successful anti-TCR combination therapies. a–d β Cells ( green ) and immune cells ( red ) were examined in islets from animals successfully treated with anti-TCR and anti-IL-6 ( b ) or with anti-TCR and anti-IL-17A ( c ) or with anti-TCR and both cytokine antibodies ( d ) after diabetes manifestation and compared to the untreated diabetic situation ( a ). Islets were immunostained for insulin ( green ) and CD68 macrophages ( red ), CD8 T cells ( red ), or γ,δ T cells (red) and counterstained with DAPI ( blue ). Erythrocytes were identified by yellow to orange colour through auto-fluorescence in the red and green channel. In each group, 40 to 80 islets in the pancreases were analysed

Article Snippet: Group 1 ( n = 6) comprised healthy, normoglycaemic IDDM rats which received 0.9% NaCl solution only, group 2 ( n = 6) comprised acutely diabetic IDDM rats which received IgG (I5381; Merck, Darmstadt, Germany; diluted with 0.9% NaCl solution) and groups 3, 4 and 5 ( n = 4 each) comprised acutely diabetic IDDM rats treated for 5 consecutive days in a monotherapy either with a rat-specific anti-TCR antibody (Clone: R73; Bio-Rad, Munich, Germany) (0.5 mg/kg b.wt. i.v.), rat-specific anti-IL-6 (0.01 mg/kg b.wt. i.v.) (Clone: MAB 561 , R&D Systems, Wiesbaden-Nordenstadt, Germany) or with a murine specific anti-IL-17A (0.1 mg/kg b.wt. i.v . ) (Clone: 17F3, eBioscience, Frankfurt, Germany ) .

Techniques: Fluorescence

Journal: BMC Medicine

Article Title: Remission of autoimmune diabetes by anti-TCR combination therapies with anti-IL-17A or/and anti-IL-6 in the IDDM rat model of type 1 diabetes

doi: 10.1186/s12916-020-1503-6

Figure Lengend Snippet: Presence of different immune cell types in the pancreatic islets

Article Snippet: Group 1 ( n = 6) comprised healthy, normoglycaemic IDDM rats which received 0.9% NaCl solution only, group 2 ( n = 6) comprised acutely diabetic IDDM rats which received IgG (I5381; Merck, Darmstadt, Germany; diluted with 0.9% NaCl solution) and groups 3, 4 and 5 ( n = 4 each) comprised acutely diabetic IDDM rats treated for 5 consecutive days in a monotherapy either with a rat-specific anti-TCR antibody (Clone: R73; Bio-Rad, Munich, Germany) (0.5 mg/kg b.wt. i.v.), rat-specific anti-IL-6 (0.01 mg/kg b.wt. i.v.) (Clone: MAB 561 , R&D Systems, Wiesbaden-Nordenstadt, Germany) or with a murine specific anti-IL-17A (0.1 mg/kg b.wt. i.v . ) (Clone: 17F3, eBioscience, Frankfurt, Germany ) .

Techniques:

Journal: BMC Medicine

Article Title: Remission of autoimmune diabetes by anti-TCR combination therapies with anti-IL-17A or/and anti-IL-6 in the IDDM rat model of type 1 diabetes

doi: 10.1186/s12916-020-1503-6

Figure Lengend Snippet: Pro-and anti-inflammatory cytokine gene expression by in situ RT-PCR in the islet immune cell infiltrate

Article Snippet: Group 1 ( n = 6) comprised healthy, normoglycaemic IDDM rats which received 0.9% NaCl solution only, group 2 ( n = 6) comprised acutely diabetic IDDM rats which received IgG (I5381; Merck, Darmstadt, Germany; diluted with 0.9% NaCl solution) and groups 3, 4 and 5 ( n = 4 each) comprised acutely diabetic IDDM rats treated for 5 consecutive days in a monotherapy either with a rat-specific anti-TCR antibody (Clone: R73; Bio-Rad, Munich, Germany) (0.5 mg/kg b.wt. i.v.), rat-specific anti-IL-6 (0.01 mg/kg b.wt. i.v.) (Clone: MAB 561 , R&D Systems, Wiesbaden-Nordenstadt, Germany) or with a murine specific anti-IL-17A (0.1 mg/kg b.wt. i.v . ) (Clone: 17F3, eBioscience, Frankfurt, Germany ) .

Techniques: Expressing, In Situ

Journal: BMC Medicine

Article Title: Remission of autoimmune diabetes by anti-TCR combination therapies with anti-IL-17A or/and anti-IL-6 in the IDDM rat model of type 1 diabetes

doi: 10.1186/s12916-020-1503-6

Figure Lengend Snippet: Cytokine pattern in serum of IDDM rats after successful anti-TCR combination therapies. Changes in protein concentrations of cytokines measured by multiplex analysis were examined after successful anti-TCR combination therapy with anti-IL-6 or anti-IL-17A alone or in triple combination after diabetes manifestation a TNF-α, b IL-1β, c IFN-γ, d IL-2, e IL-4, f IL-10, g IL-6 and h IL-17A. Results after anti-TCR combination therapy with anti-IL-6, with anti-IL-17A and in the triple fashion were compared to those from healthy controls and untreated, acutely diabetic rats. Cytokine protein concentrations (pg/ml) are expressed as mean values ± SEM; a–h the dotted lines show changes in the pro-inflammatory and anti-inflammatory cytokines compared to the normoglycaemic situation in healthy control rats. * p < 0.05 by ANOVA followed by Dunnett’s test versus untreated, diabetic controls ** p < 0.01 versus untreated, diabetic controls. Number of animals as given in Fig.

Article Snippet: Group 1 ( n = 6) comprised healthy, normoglycaemic IDDM rats which received 0.9% NaCl solution only, group 2 ( n = 6) comprised acutely diabetic IDDM rats which received IgG (I5381; Merck, Darmstadt, Germany; diluted with 0.9% NaCl solution) and groups 3, 4 and 5 ( n = 4 each) comprised acutely diabetic IDDM rats treated for 5 consecutive days in a monotherapy either with a rat-specific anti-TCR antibody (Clone: R73; Bio-Rad, Munich, Germany) (0.5 mg/kg b.wt. i.v.), rat-specific anti-IL-6 (0.01 mg/kg b.wt. i.v.) (Clone: MAB 561 , R&D Systems, Wiesbaden-Nordenstadt, Germany) or with a murine specific anti-IL-17A (0.1 mg/kg b.wt. i.v . ) (Clone: 17F3, eBioscience, Frankfurt, Germany ) .

Techniques: Multiplex Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Probiotics modulated gut microbiota suppresses hepatocellular carcinoma growth in mice

doi: 10.1073/pnas.1518189113

Figure Lengend Snippet: Probiotics retarded the tumor growth and its association with Th17 and IL-17. (A) Down-regulated IL-17 and other angiogenic factors, and up-regulated IL-10 in the two Prohep groups in 38-d samples. (B) Correspondence analysis of the qRT-PCR results of 38-d samples in four groups. (C) Tumor size variation during 38 d of monitoring with anti-IL-17 antibody. (D) Confocal images of tumor sections with IL-17 staining (blue), costained (red) with CD3 T cells (Left), CD11b macrophage (Center), and MPO neutrophils (Right). (E) Percentage of cell expressing IL-17 in CD3+, CD11b+ and MPO+ cells. (F) Frequency distribution of subpopulation of CD3+ cells in three groups. (G) Distribution of IL-17 production among different cell types. (H–L) Frequency of subpopulation of T cells in tumor: Th1 (H), TH17 (I), Th2 (J), Treg (K), and Tr1 (L). (M) Positive correlation between the Th17 proportion and tumor volume. (N) Frequency of migratory Th17 cells in the tumor section. (O) Th17 frequency in various organs measured by flowcytometry. All of the statistical tests were performed using t test between each treatment group and control group. *0.01 < P value < 0.05; **0.001 < P value < 0.01; ***P value < 0.001.

Article Snippet: To evaluate the roles of IL-17+ cells in tumor progression, IL-17 neutralization was adopted by injecting 200 μg i.p. mouse anti–IL-17 (clone 17F3, BioXcell) 1 wk before tumor inoculation.

Techniques: Quantitative RT-PCR, Staining, Expressing

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Probiotics modulated gut microbiota suppresses hepatocellular carcinoma growth in mice

doi: 10.1073/pnas.1518189113

Figure Lengend Snippet: Prohep lowers microvessel density in tumor in an IL-17-dependent manner based on the comparisons of 38-d samples. **0.001 < P value < 0.01; ***P value < 0.001 compared with the control.

Article Snippet: To evaluate the roles of IL-17+ cells in tumor progression, IL-17 neutralization was adopted by injecting 200 μg i.p. mouse anti–IL-17 (clone 17F3, BioXcell) 1 wk before tumor inoculation.

Techniques:

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: Clinical and Experimental Immunology

Article Title: Interleukin (IL)‐39 [IL‐23p19/Epstein–Barr virus‐induced 3 (Ebi3)] induces differentiation/expansion of neutrophils in lupus‐prone mice

doi: 10.1111/cei.12840

Figure Lengend Snippet: Interleukin (IL)‐39 [IL‐23p19/Epstein–Barr virus‐induced 3 (Ebi3) expanded CD11b+ cells. (a,b) Splenocytes were separated from 8‐week‐old C57BL/6 mice and cultured in vitro for 3 days in the presence of 50 ng/ml Ebi3 and IL‐39. All live cells, including large granule cells, were gated on the basis of forward‐ and side‐scatter and analysed by fluorescence activated cell sorter (FACS). The percentages of CD11c+ and CD11b+ cells (a) and statistical analysis of the percentage (b) are shown; (c–e) 400 ng/mouse p19, Ebi3 and IL‐39 were injected intravenously (i.v.) into 8‐week‐old C57BL/6 mice (six mice per group). On day 7 after injection, live lymphocyte‐sized cells were gated on the basis of forward‐ and side‐scatter and analysed by FACS. The percentages of CD138, IL‐10 or GL7‐expressing B220+ B cells (c,e) and statistical analysis of the percentage (d) are shown; (f–i) 400 ng/mouse p19 and IL‐39 were injected i.v. into 8‐week‐old C57BL/6 mice (six mice per group). On day 7 after injection, live lymphocyte‐sized cells were gated on the basis of forward‐ and side‐scatter and analysed by FACS. The percentages of CD3+ and CD4+ T cells (f) and statistical analysis of the percentage (g), IL‐10, forkhead box protein 3 (FoxP3), IL‐4, interferon (IFN)‐γ, IL‐17A‐expressing CD4+ T cells (i) and statistical analysis of the percentage (h) are shown. (j) Splenocytes were separated from 8‐week‐old C57BL/6 mice and cultured in vitro for 3 days in the presence of 50 ng/ml Ebi3 or IL‐39. Live lymphocyte‐sized cells were gated on the basis of forward‐ and side‐scatter and analysed by FACS. The percentages of IL‐10, IFN‐γ, IL‐17A‐expressing CD4 + T cells are shown. Results represent at least three independent experiments. *P < 0·05 (two‐tailed Student's t‐test). Error bars, standard error of the mean.

Article Snippet: The following antibodies were used: fluorescence‐conjugated anti‐mouse p19 (eBioscience Corp., cat. no.50‐7023‐82), Ebi3 (R&D systems, cat. no. IC18341C), IL‐12Rβ1 (BD Pharmingen, San Diego, CA, USA; 551974), IL‐12Rβ2 (Miltenyi Biotech, San Diego, CA, USA; 130‐105‐018), IL‐23R (BD Pharmingen; 551974), IL‐27Ra (R&D Systems, Minneapolis, MA, USA; 263503), gp130 (eBioscience;17‐1302), B220 (eBioscience; RA3‐6B2), CD19 (eBioscience; MB19‐1), GL7 (eBioscience; GL‐7), CD138 (eBioscience; DL‐101), IL‐10 (eBioscience; JES5‐16E3), CD3 (eBioscience; 145‐2C11), CD4 (eBioscience; GK1.5), CD11b (eBioscience; M1/70), CD11c (eBioscience; N418), IL‐4 (eBioscience; 11B11), IL‐17A (eBioscience; 17F3), forkhead box protein 3 (FoxP3) (eBioscience; NRRF‐30), interferon (IFN)‐γ (eBioscience; XMG1.2), Gr‐1 (eBioscience; RB6‐8C5), BAFF (Pierce, MA, USA; 125955), phosphor signal transducer and activator of transcription‐1 (pSTAT‐1) (Santa Cruz Biotech; sc‐8394) and pSTAT‐3 (Santa Cruz Biotech; sc‐8059) antibodies.

Techniques: Cell Culture, In Vitro, Fluorescence, Injection, Expressing, Two Tailed Test