human il 17a f  (R&D Systems)

Journal: bioRxiv

Article Title: LL-37 modulates IL-17A/F-mediated airway inflammation by selectively suppressing Lipocalin-2

doi: 10.1101/2024.09.03.610924

Figure Lengend Snippet: HBEC-3KT cells were stimulated with IL-17A/F (50 ng/mL) for 24 h. (A) Cell lysates from cells stimulated with IL- 17A/F and unstimulated cells (14 µg total protein per sample), obtained from five independent experiments (n=5), were independently probed using the high-content aptamer-based proteomic array. Pairwise differential analysis was conducted on normalized log2 protein expression values, and Welch’s t-test with a cutoff of p<0.05 was used to select protein abundance changes that were significantly altered in response to IL-17A/F. (B) TC supernatant collected from cells 24 h post- stimulation was examined for the abundance of LCN-2, Elafin, and GROα, by ELISA. Each dot represents an independent experiment (n=5), and bars show the median and min-max range. Repeated measures one-way ANOVA with Fisher’s least significant difference test was used for statistical analysis ( *p≤0.05, ***p≤0.005 ).

Article Snippet: Recombinant human cytokines IL-17A/F (carrier free) was obtained from R&D Systems (Oakville, ON, CA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: LL-37 modulates IL-17A/F-mediated airway inflammation by selectively suppressing Lipocalin-2

doi: 10.1101/2024.09.03.610924

Figure Lengend Snippet: HBEC-3KT cells were stimulated with either LL-37, citLL-37 or sLL- 37 (0.25 μM), in the presence and absence of IL-17A/F (50 ng/mL), for 24 h. TC supernatants collected from seven independent experiments (n=7) were examined by ELISA for the abundance LCN-2, Elafin and GROα. Y-axis represents % change compared to paired unstimulated controls. Each dot represents an independent experiment, and bars show the median and min-max range. Each dot is reported as percent (%) change compared to unstimulated cells where % change = ((treatment – control) / control) x 100%. Repeated measures one-way ANOVA with Fisher’s least significant difference test was used for statistical analysis ( *p≤0.05, ****p≤0.0001 ).

Article Snippet: Recombinant human cytokines IL-17A/F (carrier free) was obtained from R&D Systems (Oakville, ON, CA).

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: bioRxiv

Article Title: LL-37 modulates IL-17A/F-mediated airway inflammation by selectively suppressing Lipocalin-2

doi: 10.1101/2024.09.03.610924

Figure Lengend Snippet: HBEC- 3KT cells were stimulated with IL-17A/F (50 ng/mL) + TNFα (20 ng/mL) for 24 h. TC supernatants were collected and used in the bottom chamber of transwell plates in a neutrophil migration assays, wherein neutrophils isolated from human blood were used in the upper chamber of the transwell plates. Cell culture medium spiked with human recombinant IL-8 (30 ng/mL) was used as a positive control for neutrophil migration. Results are shown as boxplots with the median line and IQR, and whiskers show minimum and maximum values. Each data point represents an independent experimental replicate with TC supernatant (n=3), using neutrophil isolated from one donor. Each dot represents the average number of neutrophils that traversed the membrane within two hours in each experiment, and bars show the mean and SEM. Repeated measures one-way ANOVA with Fisher’s least significant difference test was used for statistical analysis ( *p≤0.05, **p≤0.01 ).

Article Snippet: Recombinant human cytokines IL-17A/F (carrier free) was obtained from R&D Systems (Oakville, ON, CA).

Techniques: Migration, Isolation, Cell Culture, Recombinant, Positive Control, Membrane

Journal: bioRxiv

Article Title: LL-37 modulates IL-17A/F-mediated airway inflammation by selectively suppressing Lipocalin-2

doi: 10.1101/2024.09.03.610924

Figure Lengend Snippet: HBEC-3KT cells (n=5) were stimulated with either LL-37, citLL-37 or sLL-37 (0.25 μM), in the presence and absence of IL-17A/F (50 ng/mL). mRNA was isolated after (A) 3 h and (B) 6 h, and the mRNA abundance of NGAL2 , NFKBIZ and CEBPB was examined by qRT-PCR. Fold changes (Y-axis) for each gene candidate was normalized to 18S RNA, and compared to unstimulated cells normalized to 1, using the comparative ΔΔCt method. Each data point represents an independent experimental replicate and bars show the mean and SEM. Fisher’s LSD test for one-way ANOVA was used to determine statistical significance ( *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 ).

Article Snippet: Recombinant human cytokines IL-17A/F (carrier free) was obtained from R&D Systems (Oakville, ON, CA).

Techniques: Isolation, Quantitative RT-PCR

Journal: bioRxiv

Article Title: LL-37 modulates IL-17A/F-mediated airway inflammation by selectively suppressing Lipocalin-2

doi: 10.1101/2024.09.03.610924

Figure Lengend Snippet: Human PBEC obtained from three independent donors (N=3) were stimulated with LL-37, citLL-37, or sLL-37 (0.25 μM), in the presence and absence of IL-17A/F (50 ng/mL). Cell lysates (25 μg total protein per sample) was used to examine the abundance of Regnase-1 (A) 30 minutes and (B) 24 h post-stimulation, by Western blots. (C) The abundance of p-IKKα/β (S176/180) and NF-κB p65, 30 minutes post-stimulation, by Western blots. (D) Human PBEC obtained from three independent donors (N=3) were stimulated with LL- 37, citLL-37, or sLL-37 (0.25 μM), in the presence and absence of IL-17A/F (50 ng/mL) and TNF-α (20 ng/mL). The abundance of LCN-2, 24 h post-stimulation, by ELISA. Y-axis represents % change compared to paired unstimulated cells from each donor. Each dot represents an independent experiment, and bars show the mean and SEM. Each dot is reported as percent (%) change compared to unstimulated PBEC where % change = ((treatment – control) / control) x 100%. Repeated measures one-way ANOVA with Fisher’s least significant difference test was used for statistical analysis ( *p≤0.05 ).

Article Snippet: Recombinant human cytokines IL-17A/F (carrier free) was obtained from R&D Systems (Oakville, ON, CA).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Control

Journal: bioRxiv

Article Title: LL-37 modulates IL-17A/F-mediated airway inflammation by selectively suppressing Lipocalin-2

doi: 10.1101/2024.09.03.610924

Figure Lengend Snippet: (A) HBEC-3KT cells (n=6) and (B) human PBEC (N=3 independent donors, n=2 technical replicate each) were stimulated with LL-37, citLL-37 or sLL-37 (0.25 μM), in the presence/absence of IL-17A/F (50 ng/mL), for 24 h. TC supernatants were examined for the abundance of CCL20 by ELISA. Y-axis represents % change compared to paired unstimulated cells for each replicate. Each dot represents an independent experiment, and bars show the median and min-max range. Each dot is reported as percent (%) change compared to unstimulated control bronchial epithelial cells where % change = ((treatment – control) / control) x 100%. Repeated measures one-way analysis of variance with Fisher’s least significant difference test was used for statistical analysis ( **p≤0.001, ***p≤0.005, ****p≤0.0001 ).

Article Snippet: Recombinant human cytokines IL-17A/F (carrier free) was obtained from R&D Systems (Oakville, ON, CA).

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: bioRxiv

Article Title: LL-37 modulates IL-17A/F-mediated airway inflammation by selectively suppressing Lipocalin-2

doi: 10.1101/2024.09.03.610924

Figure Lengend Snippet: Female and male BALB/c mice (8-10 weeks; N≥4 per group) were challenged (i.n.) with either saline, 25 μg of HDM protein extract (35 μL of 7 µg/mL saline per mouse) with or without 1 μg LPS (35 μL of 0.03 µg/mL saline per mouse), or LPS alone, once daily for 3 days (days 0 to 2), rested for 4 days followed by HDM allergen recall challenge once daily for 8 consecutive days (as detailed in ). Abundance of (A) CRAMP and (B) IL-17A/F in bronchoalveolar lavage (BAL) was assessed by ELISA, 24 h after the last HDM challenge. (C) Abundance of LCN-2 was assessed in the BALF and lung tissue lysates by ELISA. Bars show median and IQR, whiskers show minimum and maximum points, + denotes average. Statistical analysis was determined by one-way ANOVA with Fisher’s LSD test ( *p≤0.05, **p≤0.001, ***p≤0.005, ****p≤0.0001 ). HDM, house dust mite; LPS, lipopolysaccharide.

Article Snippet: Recombinant human cytokines IL-17A/F (carrier free) was obtained from R&D Systems (Oakville, ON, CA).

Techniques: Saline, Enzyme-linked Immunosorbent Assay

Journal: Biomedicines

Article Title: Novel Biotherapeutics Targeting Biomolecular and Cellular Approaches in Diabetic Wound Healing

doi: 10.3390/biomedicines11020613

Figure Lengend Snippet: Currently available biological therapeutic agents.

Article Snippet: Ixekizumab , Humanized anti-IL-17A and17A/F ImmunoglobulinG4 , Taltz ® (LY2439821 Eli Lily & Co.) , America.

Techniques: Recombinant, Derivative Assay