human tslp, il-6, and ccl26 elisa development abts kits  (PeproTech)

Journal: Frontiers in Immunology

Article Title: Staphylococcus aureus δ-toxin present on skin promotes the development of food allergy in a murine model

doi: 10.3389/fimmu.2023.1173069

Figure Lengend Snippet: Murine keratinocytes released IL-1α in response to stimulation with δ-toxin. Murine keratinocytes were stimulated with different concentrations of δ-toxin for 2 or 24 h, as indicated. (A, B) Concentrations of (A) IL-1α and (B) TSLP in the culture supernatants. (C) Relative expression levels of mRNA of IL-1α and IL-36α in the δ-toxin-stimulated keratinocytes. (D) The percentage of dead cells. (A–D) Data are representative of three independent experiments. Means ± SD have been plotted. * P < 0.05 or ** P < 0.01.

Article Snippet: ELISA kits for IL-4, IL-13, IL-33, IL-1α, IL-1β, IL-25, and TSLP (R&D Systems), mast cell protease-1 (MCPT-1) (eBioscience), and high mobility group box 1 (HMGB1) (Promega) were used to measure their concentrations in serum, culture supernatants, and skin tissue homogenates.

Techniques: Expressing

Journal: Nature Communications

Article Title: Context-dependent function of TSLP and IL-1β in skin allergic sensitization and atopic march

doi: 10.1038/s41467-022-32196-1

Figure Lengend Snippet: a RNAscope in situ hybridization for IL-1β mRNA in non-treated (NT), LMP_30μm and LMP_91 μm ears at 48 h after the microporation. Dashed lines indicate the dermal/epidermal junction. b ELISA measurement of IL-1β protein levels in ears at 48 h after LMP_30 μm and 91 μm ( n = 5, 12, 6 mice). c Experimental protocol. HDM with or without IL-1β was applied on LMP_30μm ears (e.c. HDM ± IL-1β), at day (D) 0, D3, D7 and D10. Mice were intranasally (i.n.) challenged with HDM every day from D9 to D12 to induce allergic asthma, and analysed at D13. d ELISA measurement of TSLP protein levels in 30 μm-LMP ears co-administrated with recombinant IL-1β or PBS ( n = 4 mice). e Hematoxylin & eosin (H&E) staining and immunohistochemistry (IHC) staining for MBP or MCPT8 on ear sections. Black arrows point to one of the positive signals. Scale bar = 50 μm for all pictures. f Comparison of CXCR5 + PD1 + Tfh cells, GL7 + Fas + GC B cells, IgE + B and IgG1 + B cells in EDLNs ( n = 4, 5, 3, 3, 3, 4 mice). g Serum levels of HDM-specific IgG1 and IgE in HDM-treated mice ( n = 5, 5, 6, 4 mice). Graphs in b , d , f show mean ± SEM. Two-sided Student’s t -test. Graphs in g show median. Two-sided Mann–Whitney rank sum test. All data are representative of two independent experiments with similar results. Source data are provided as a Source Data file.

Article Snippet: TSLP and IL-1β levels in skin extracts were determined using the DuoSet ELISA Development Kits (R&D Systems, Minneapolis, Minn, Cat No. DY555 for TSLP, Cat No. DY401 for IL-1β).

Techniques: RNAscope, In Situ Hybridization, Enzyme-linked Immunosorbent Assay, Recombinant, Staining, Immunohistochemistry, Comparison, MANN-WHITNEY

Journal: Nature Communications

Article Title: Context-dependent function of TSLP and IL-1β in skin allergic sensitization and atopic march

doi: 10.1038/s41467-022-32196-1

Figure Lengend Snippet: a Experimental protocol. Wildtype Balb/c mice were intraperitoneally (i.p.) injected with PBS, NIMP-R14 or anti-Ly6G antibody (Ab) at day (D) -1 and D2. Mice were d.c. sensitized with HDM on LMP_91μm ears at D0 and D3 or non-sensitized. All mice were intranasally (i.n.) challenged with HDM from D10 to D13 and analysed at D14. b Total cell number and differential cell counting in BAL fluid ( n = 4, 4, 4, 3 mice). c Relative mRNA levels of genes in BAL cells ( n = 4, 4, 3, 4 mice). d Lung sections were stained with hematoxylin-eosin (H&E) for histological analyses or Periodic Acid Schiff (PAS) for goblet cell hyperplasia analyses. B: bronchiole. V: blood vessel. Bar = 250 μm for all pictures. e Serum level of HDM-specific IgE measured by ELISA ( n = 3 mice). Graphs in b , c show mean ± SEM, One-way ANOVA test. Graph in e marks median, two-sided Mann–Whitney rank sum test. All data are representative of two independent experiments with similar result. Source data are provided as a Source Data file.

Article Snippet: TSLP and IL-1β levels in skin extracts were determined using the DuoSet ELISA Development Kits (R&D Systems, Minneapolis, Minn, Cat No. DY555 for TSLP, Cat No. DY401 for IL-1β).

Techniques: Injection, Cell Counting, Staining, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Nature Communications

Article Title: Context-dependent function of TSLP and IL-1β in skin allergic sensitization and atopic march

doi: 10.1038/s41467-022-32196-1

Figure Lengend Snippet: a Experimental protocol. Wildtype Balb/c mice were intraperitoneally (i.p.) injected with PBS or NIMP-R14 antibody at day (D) -1 and D2. Mice were d.c. sensitized with HDM ± IL-1β on LMP_91μm ears at D0 and D3. All mice were intranasally (i.n.) challenged with HDM from D9 to D12 and analysed at D13. b Total cell number and differential cell counting in BAL fluid ( n = 5, 4, 5 mice). One-way ANOVA test. c Relative RNA levels of genes in BAL cells ( n = 5, 4, 4 mice). One-way ANOVA test. d Lung sections were stained with hematoxylin-eosin (H&E) or Periodic Acid Schiff (PAS). B: bronchiole. V: blood vessel. Bar = 250 μm for all pictures. e Lung resistance (R L ) at the baseline (aerosol of PBS) and in response to aerosolized methacholine (Mch; 50 mg/ml), measured by FlexiVent system ( n = 4, 5, 4, 5 mice). Two-sided Student’s t test. f Serum level of HDM-specific IgE and IgG1 measured by ELISA ( n = 5, 4, 5 mice). Two-sided Mann–Whitney rank sum test. NS, non-sensitized. Data are representative of 2 ( b – d , f ) or 1 ( e ) independent experiments with similar results. Source data are provided as a Source Data file.

Article Snippet: TSLP and IL-1β levels in skin extracts were determined using the DuoSet ELISA Development Kits (R&D Systems, Minneapolis, Minn, Cat No. DY555 for TSLP, Cat No. DY401 for IL-1β).

Techniques: Injection, Cell Counting, Staining, Aerosol, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Biomedicines

Article Title: CCL4 Functions as a Biomarker of Type 2 Airway Inflammation

doi: 10.3390/biomedicines10081779

Figure Lengend Snippet: Epithelial cell-derived cytokines and chemokines levels during eosinophilic inflammation. ( A , B ) Innate immune response-associated cytokine mRNA in nasal polyps from patients with ECRS ( A ) and BEAS-2B cells co-incubated overnight with purified peripheral blood eosinophils or recombinant eosinophil peroxidase (EPX, 10 µg/mL) for 72 h ( B ). ( C ) TSLP release from BEAS-2B cells incubated with EPX for 48 h. ( D , E ) Eosinophil-recruiting chemokines in BEAS-2B cells incubated with TSLP (10 ng/mL) for 48 h, followed by overnight stimulation with EPX (10 µg/mL). CCL4, CCL5, CCL11, and CCL26 mRNA levels ( D ) and CCL4 release ( E ) were evaluated. ( F ) TSLP receptor (TSLPR) mRNA levels in BEAS-2B cells co-incubated with EPX for 48 h. Values in (( A , B , D , F ) represent ratios to control (uncinate process tissue; UT from the same patients or non-treatment; vehicle) and values in all panels represent the mean ± standard error of the mean from four experiments; # p < 0.05, ## p < 0.01 (vs. UT or vehicle), * p < 0.05 (as shown between the two groups).

Article Snippet: CCL4 and TSLP levels in cell culture supernatants were measured using MIP-1β/CCL4 and TSLP ELISA kits (R&D Systems, Minneapolis, MN, USA).

Techniques: Derivative Assay, Incubation, Purification, Recombinant, Control

Journal: Biomedicines

Article Title: CCL4 Functions as a Biomarker of Type 2 Airway Inflammation

doi: 10.3390/biomedicines10081779

Figure Lengend Snippet: TLR3 ligand-mediated CCL4 release. ( A ) CCL4 release from BEAS-2B cells stimulated overnight with TLR ligands (zymosan, poly I:C, LPS, imiquimod), and the PAR2 agonist (2-Furoyl-LIGRLO-amide). ( B ) CCL4 release from BEAS-2B cells co-incubated overnight with TSLP and poly I:C. ( C ) TLR3 mRNA levels in BEAS-2B cells incubated with TSLP for 48 h, followed by overnight stimulation with EPX (10 µg/mL). Values represent the mean ± standard error of the mean from four ( A , B ) or three ( C ) experiments; # p < 0.05, ## p < 0.01 (vs. vehicle in each group), * p < 0.05, ** p < 0.01 (as shown between the two groups).

Article Snippet: CCL4 and TSLP levels in cell culture supernatants were measured using MIP-1β/CCL4 and TSLP ELISA kits (R&D Systems, Minneapolis, MN, USA).

Techniques: Incubation