il 17 a  (R&D Systems)

Journal: Journal of Nanobiotechnology

Article Title: Mitochondria-targeted nanozyme system for psoriasis treatment

doi: 10.1186/s12951-026-04068-z

Figure Lengend Snippet: Therapeutic effects of CMH@lip@Res-TCeO2 on IMQ-induced psoriasiform skin inflammation. ( A - B ) Macroscopic observation of dorsal skin lesions in mice, evaluating erythema, scaling, and inflammatory infiltration; ( C ) H&E staining to assess histopathological features and changes in epidermal thickness (scale bar: 50 μm); ( D ) Ly6G IF staining to evaluate neutrophil infiltration in skin tissues (scale bar: 50 μm); ( E ) Colorimetric assay to measure MPO activity in skin tissues; ( F - G ) Detection of ROS and MDA levels using fluorescent probes and evaluation of CAT, SOD, and GSH activities using colorimetric assays; ( H ) IHC analysis of Ki67 expression as a proliferation marker in skin tissues (scale bar: 50 μm); ( I ) ELISA quantification of IL-23, IL-1β, IL-17 A, and TNF-α levels in skin homogenates. Each group included six animals. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: Cytokine levels in cell culture supernatants and tissue homogenates were measured using ELISA kits for IL-1β (DY401), IL-6 (M6000B), TNF-α (MTA00B), IL-17 A (DY5390), and IL-23 (M2300) (R&D Systems, USA).

Techniques: Staining, Colorimetric Assay, Activity Assay, Expressing, Marker, Enzyme-linked Immunosorbent Assay

Journal: PLOS One

Article Title: Clinical proof of concept for small molecule mediated inhibition of IL-17 in psoriasis

doi: 10.1371/journal.pone.0341049

Figure Lengend Snippet: (A) Inhibition of IL-17 bioactivity as secreted by human TH17 cells and assayed by a reporter cell assay. Dotted lines represent positive and negative assay controls. 1 representative of 7 experiments with different donors is shown. The error bars represent the SEM. (B, C) Evaluation of DC-806 in rat CIA. Rats were randomized on Day 11 and dosed as indicated. Daily measurements of ankle thickness (B) and terminal measurement of footpad weights (C) were used as efficacy readouts. (D) Preclinical evaluation of DC-806 serum levels. PK sampling for exposure determination was performed on Days 11, 16, at 1 and 12 hours post dose. The dotted lines represent the uncorrected IC 50 for rat IL-17AA and rat IL-17AF as listed in . The dashed line represents the LLQ of the quantification assay. All error bars represent the SEM. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Abbreviations : anti-F, anti-IL-17F; BID, twice daily; CIA, collagen-induced arthritis; Dex, dexamethasone; IC 50 , half-maximal inhibitory concentration; IL-17, interleukin-17; LLQ, lower limit of quantification; PK, pharmacokinetic; QD, once daily; SEM, standard error of mean; TH17, T-helper 17.

Article Snippet: To determine cellular potency, recombinant human (h)IL-17AA (Genscript cat# Z03228), hIL-17AF (R&D cat# 5837-IL-010), hIL-17FF (R&D Systems 1335-IL), rat IL-17AA (R&D systems 8410-IL-025), and rat IL-17AF (R&D systems 9340-IL-025) were used for in vitro stimulation at the concentrations listed in .

Techniques: Inhibition, Sampling, Concentration Assay

Journal: PLOS Pathogens

Article Title: Mucosal immunization with the lung Lactobacillus -derived amphiphilic exopolysaccharide adjuvanted recombinant vaccine improved protection against P . aeruginosa infection

doi: 10.1371/journal.ppat.1012696

Figure Lengend Snippet: (A) Experimental design and timeline of prime and booster regimen of PBS, rPcrV, CTB+rPcrV and EPS301@rPcrV vaccination (i.n.) and challenge. rPcrV-specific IgA titers in lung homogenate and IgG titers in serum were determined by ELISA before each round of immunization and every 14 days after the primary immunization. (B) The graph line shown the evolution of IgA titers in lung homogenate post immunization for each immunized group of mice. *p < 0.05; **p < 0.01; ***p < 0.001 for comparison with CTB+rPcrV immunized mice (n = 8). (C) The graph line shown the evolution of IgG titers in serum post immunization for each immunized group of mice. *p < 0.05 for comparison with rPcrV alone immunized mice (n = 8). (D) Mice were inoculated with 40 μl bacterial slurry (1×10 9 CFUs of P . aeruginosa PAO1) via left nostril on day 112 post booster vaccination and held upright for 1 min. Representative survival rates from two independent experiments are shown (n = 8–10). The data for survival test were analyzed by Wilcoxon log-rank survival test (*p < 0.05; **p < 0.01). (E) Mice were inoculated with 40 μl bacterial slurry (lower dose, 1×10 7 CFUs of P . aeruginosa PAO1) as above. At 12 h post challenge, the numbers of bacteria in lungs, BALF and spleens were counted (n = 5–8). (F) Mice were inoculated with the lower dose of PAO1 as above. At 12 h post challenge, histological evaluation of lung sections by light microscopy. Lung specimens were fixed, sectioned, and stained with H&E (n = 3–5). Data are presented as means ± SEM. Significant differences were calculated with One- or Two-way ANOVA followed by Tukey’s multiple comparisons test. ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: To define the IL-17A and IFN-γ levels in lung and spleen tissue homogenate, the murine bioplex ELISA kits (R&D SYSTEMS Mouse IL-17A/F Heterodimer DuoSet ELISA DY5390 and R&D SYSTEMS Mouse IFN-gamma DuoSet ELISA DY485) were used according to the manufacturer’s recommendations.

Techniques: Enzyme-linked Immunosorbent Assay, Comparison, Bacteria, Light Microscopy, Staining

Journal: PLOS Pathogens

Article Title: Mucosal immunization with the lung Lactobacillus -derived amphiphilic exopolysaccharide adjuvanted recombinant vaccine improved protection against P . aeruginosa infection

doi: 10.1371/journal.ppat.1012696

Figure Lengend Snippet: Mice (n = 3-5/group) were immunized (i.n.) twice 14 days apart with rPcrV, CTB+ rPcrV or EPS301@rPcrV, with animals receiving PBS served as controls. Mice were inoculated with 40 μl bacterial slurry (lower dose, 1×10 7 CFUs of P . aeruginosa PAO1) as above. Vaccinated mice were sacrificed at 12 hours post-challenge on day 7 or day 112 after the second vaccination lung tissue were prepared. Number of IFN-γ + CD4 + T cells, IL-17A + CD4 + T cells (A), IFN-γ + γδ + T cells, IL-17A + γδ + T cells (B) in lung at 12 hours post-challenge on day 7 after the second vaccination were estimated by intracellular cytokine. Number of IFN-γ + CD4 + T cells, IL-17A + CD4 + T cells (C), IFN-γ + γδ + T cells, IL-17A + γδ + T cells (D) in lung at 12 hours post-challenge on day 112 after the second vaccination were estimated by intracellular cytokine. IFN-γ and IL-17 levels, determined by ELISA in a supernatant of lung tissue homogenate were analyzed. The IFN-γ levels and IL-17A levels (E) in lung at 12 hours post-challenge on day 7 and day 112 after the second vaccination were determined by ELISA. Data are presented as means ± SEM. Significant differences were calculated with One-way ANOVA followed by Tukey’s multiple comparisons test. *p < 0.05; **p < 0.01; ***p < 0.001 for comparison with EPS301@rPcrV immunized mice.

Article Snippet: To define the IL-17A and IFN-γ levels in lung and spleen tissue homogenate, the murine bioplex ELISA kits (R&D SYSTEMS Mouse IL-17A/F Heterodimer DuoSet ELISA DY5390 and R&D SYSTEMS Mouse IFN-gamma DuoSet ELISA DY485) were used according to the manufacturer’s recommendations.

Techniques: Enzyme-linked Immunosorbent Assay, Comparison

Journal: PLOS Pathogens

Article Title: Mucosal immunization with the lung Lactobacillus -derived amphiphilic exopolysaccharide adjuvanted recombinant vaccine improved protection against P . aeruginosa infection

doi: 10.1371/journal.ppat.1012696

Figure Lengend Snippet: (A) Comparative analysis of opsonophagocytosis against PAO1 using antisera from differently immunized mice. (B) Timeline of passive transfer and P . aeruginosa -induced pneumonia model. (C) Passive transfer of immunized and non-immunized mice was evaluated by i.v. injection of pooled serum (100 μl) to naïve C57BL/6 mice (n = 10/group). 24 hours after serum transfer, mice were inoculated with 40 μl bacterial slurry (1×10 9 CFUs of P . aeruginosa PAO1) via intranasal route (i.n.). Survival of mice from PBS, rPcrV, rPcrV+CTB and EPS301@rPcrV groups were observed for 48 h. The data for survival test were analyzed by Wilcoxon log-rank survival test (ns, not significant). (D) A lower dose of bacterial slurry (1×10 7 CFUs of P . aeruginosa PAO1) was inoculated to recipient mice via intranasal route (i.n.). Bacterial loads in lung and BALF were detected at 12 h post infection. (E) 12 hours post-challenge, lung tissue was prepared. IL-17A and IFN-γ levels determined by ELISA in a supernatant of lung tissue homogenate were analyzed. (F) Timeline of adoptive transfer and P . aeruginosa -induced pneumonia model. Lung and splenic CD3 + CD4 + T cells from EPS301@rPcrV–vaccinated congenic mice were purified on day 7 after second vaccination. CD3 + CD4 + T cells (5×10 4 ) were intravenously transferred into naïve C57BL/6 mice. (G) 24 hours after adoptive transfer, mice were inoculated with 40 μl bacterial slurry (1×10 9 CFUs of P . aeruginosa PAO1) via intranasal route (i.n.). Survival of mice were observed for 48 h. Survival after P . aeruginosa challenge (data were pooled from two independent experiments (n = 8–10). The data for survival test were analyzed by Wilcoxon log-rank survival test (***p < 0.001). (H) A lower dose of bacterial slurry (1×10 7 CFUs of P . aeruginosa PAO1) was inoculated to recipient mice via intranasal route (i.n.). Bacterial loads in lung and BALF were detected at 12 h post infection. (I) 12 hours post-challenge, lung tissue was prepared. IL-17A and IFN-γ levels determined by ELISA in a supernatant of lung tissue homogenate were analyzed. (J) Coexpression of CD44 and CD69 on non-stimulated lung CD4 + or γδ + T cells (gated in CD45 + CD4 + T cells). (K) FTY720 was initially dissolved in DMSO to create a 50 mg/mL stock solution. This stock solution was subsequently diluted with saline for intraperitoneal administration at a dosage of 30 μg per mice, and with distilled water for administration via stomach intubation at a dosage of 50 μg per mice. A lower dose of bacterial slurry (1×10 7 CFUs of P . aeruginosa PAO1) was inoculated to FTY720 treated and FTY720 non-treated mice via intranasal route (i.n.). Bacterial loads in lung were detected at 12 h post P . aeruginosa infection. Data are presented as means ± SEM. Significant differences were calculated with One-way ANOVA followed by Tukey’s multiple comparisons test (D, E, H and L), Mann-Whitney U test (J) or unpaired t test (K). *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: To define the IL-17A and IFN-γ levels in lung and spleen tissue homogenate, the murine bioplex ELISA kits (R&D SYSTEMS Mouse IL-17A/F Heterodimer DuoSet ELISA DY5390 and R&D SYSTEMS Mouse IFN-gamma DuoSet ELISA DY485) were used according to the manufacturer’s recommendations.

Techniques: Injection, Infection, Enzyme-linked Immunosorbent Assay, Adoptive Transfer Assay, Purification, Saline, MANN-WHITNEY

Journal: PLOS Pathogens

Article Title: Mucosal immunization with the lung Lactobacillus -derived amphiphilic exopolysaccharide adjuvanted recombinant vaccine improved protection against P . aeruginosa infection

doi: 10.1371/journal.ppat.1012696

Figure Lengend Snippet: TCR δ-deficient mice were vaccinated with EPS301@rPcrV and survival, bacterial loads and the lung pathology were detected on day 7 post second vaccination. Mice were inoculated with 40 μl bacterial slurry (1×10 9 CFUs of P . aeruginosa PAO1) via intranasal route (i.n.). (A) The number and percentage of Th 17 cells in WT and TCR δ-deficient mice before immunization, after immunization and post infection. (B) Survival of mice were observed for 48 h. Mice were inoculated with 40 μl bacterial slurry (1×10 7 CFUs of P . aeruginosa PAO1) via intranasal route (i.n.). The data for survival test were analyzed by Wilcoxon log-rank survival test (***p < 0.001). (C) Bacterial burdens of mice were counted at 12 h post infection. (D) Histological evaluation of lung sections by light microscopy. Lung specimens were fixed, sectioned, and stained with H&E (n = 3–5). (E) Timeline of adoptive transfer and P . aeruginosa -induced pneumonia model. Lung and splenic CD3 + γδ T cells from EPS301@rPcrV–vaccinated congenic mice were purified on day 7 after second vaccination. CD3 + γδ T cells (1 × 10 4 ) were intravenously transferred into naïve C57BL/6 mice. (F) 24 hours after adoptive transfer, mice were inoculated with 40 μl bacterial slurry (1×10 9 CFUs of P . aeruginosa PAO1) via intranasal route (i.n.). Survival of mice were observed for 48 h. Survival after P . aeruginosa challenge (data were pooled from two independent experiments (n = 8–10). The data for survival test were analyzed by Wilcoxon log-rank survival test (**p < 0.01, ***p < 0.001). A lower dose of bacterial slurry (1×10 7 CFUs of P . aeruginosa PAO1) was inoculated to recipient mice via i.n.. (G) Bacterial loads in lung and BALF were detected at 12 h post infection. (H) 12 hours post-challenge, lung tissue was prepared. IL-17A and IFN-γ levels determined by ELISA in a supernatant of lung tissue homogenate were analyzed. Data are presented as means ± SEM. Significant differences were calculated with unpaired t test (A) and One-way ANOVA followed by Tukey’s multiple comparisons test (C, G and H). ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: To define the IL-17A and IFN-γ levels in lung and spleen tissue homogenate, the murine bioplex ELISA kits (R&D SYSTEMS Mouse IL-17A/F Heterodimer DuoSet ELISA DY5390 and R&D SYSTEMS Mouse IFN-gamma DuoSet ELISA DY485) were used according to the manufacturer’s recommendations.

Techniques: Infection, Light Microscopy, Staining, Adoptive Transfer Assay, Purification, Enzyme-linked Immunosorbent Assay

Journal: PLOS Pathogens

Article Title: Mucosal immunization with the lung Lactobacillus -derived amphiphilic exopolysaccharide adjuvanted recombinant vaccine improved protection against P . aeruginosa infection

doi: 10.1371/journal.ppat.1012696

Figure Lengend Snippet: (A) Experimental design and timeline of prime and booster regimen to IL-17A-deficient, IFN-γ-deficient and WT mice of EPS301-adjuvanted vaccination (i.n.) and challenge. (B) 7 days post booster vaccination, level of IgG in serum was detected by ELISA. (C) 7 days post booster vaccination, level of IgA in lung was detected by ELISA. (D) IL-17A-deficient, IFN-γ-deficient and WT mice were inoculated with 40 μl bacterial slurry (1×10 9 CFUs of P . aeruginosa PAO1) via left nostril on day 7 post booster vaccination and held upright for 1 min. Representative survival rates from two independent experiments are shown (n = 8–10). The data for survival test were analyzed by Wilcoxon log-rank survival test (**p < 0.01, ***p < 0.001). (E) Mice were inoculated with 40 μl bacterial slurry (lower dose: 1×10 7 CFUs of P . aeruginosa PAO1) as above. At 12 h, the numbers of bacteria in lung and BALF were counted (n = 5–8). (F) Mice were inoculated with the lower dose of PAO1 as above. At 12 h post challenge, histological evaluation of lung sections by light microscopy. Lung specimens were fixed, sectioned, and stained with H&E (n = 3–5). Data are presented as means ± SEM. Significant differences were calculated with One-way ANOVA followed by Tukey’s multiple comparisons test. ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001).

Article Snippet: To define the IL-17A and IFN-γ levels in lung and spleen tissue homogenate, the murine bioplex ELISA kits (R&D SYSTEMS Mouse IL-17A/F Heterodimer DuoSet ELISA DY5390 and R&D SYSTEMS Mouse IFN-gamma DuoSet ELISA DY485) were used according to the manufacturer’s recommendations.

Techniques: Enzyme-linked Immunosorbent Assay, Bacteria, Light Microscopy, Staining