Journal: Scientific Reports

Article Title: Characterization of local and circulating bovine γδ T cell responses to respiratory BCG vaccination

doi: 10.1038/s41598-019-52565-z

Figure Lengend Snippet: Cellular immune responses in the airways and peripheral blood of BCG-vaccinated calves. ( A ) Peripheral blood was collected 4-, 8-, 12- and 16-weeks after vaccination from control (n = 7) or BCG–vaccinated animals (n = 7). PBMC were isolated and stimulated for 6 days with PPD-b. Control wells remained unstimulated. ( B ) BAL were collected at 4-, 12- and 16-weeks after vaccination. BAL cells were stimulated for 72 hours in vitro with PPD-b. Control wells remained unstimulated. Cell culture supernatants were collected from BAL and PBMC cultures and analyzed by commercial ELISA kit for IFNγ (upper panel) and IL-17 (lower panel). Data represent means ± SEM. *p < 0.05, **p < 0.01 ***p < 0.001 as determined by Student’s t test. This figure was previously published and is duplicated with permission.

Article Snippet: Briefly, 96-well ELISPOT plates (Millipore) were coated at 4 °C overnight with an anti-bovine IFNγ capture mAb or IL-17A capture mAb (both from Kingfisher Biotech, Inc., St. Paul, MN), followed by a blocking step in cRPMI, for 2 h at 37 °C, 5% CO 2 .

Techniques: Control, Isolation, In Vitro, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Characterization of local and circulating bovine γδ T cell responses to respiratory BCG vaccination

doi: 10.1038/s41598-019-52565-z

Figure Lengend Snippet: Cellular immune responses in lung-draining lymph nodes of BCG-vaccinated calves. Lung-draining tracheobronchial ( A ) and mediastinal lymph nodes ( B ), were collected from control (n = 7) or BCG–vaccinated animals (n = 7) during necropsy at 16 weeks after vaccination. Cells were isolated as described in Materials & Methods and restimulated for 6 days with PPD-b. Control wells remained unstimulated. Cell culture supernatants were collected from cultures and analyzed by commercial ELISA kit for IFNγ (upper panel) and IL-17 (lower panel). Data represent means ± SEM. **p < 0.01 ***p < 0.001 ****p < 0.001 as determined by Student’s t test. This figure was previously published and is duplicated with permission.

Article Snippet: Briefly, 96-well ELISPOT plates (Millipore) were coated at 4 °C overnight with an anti-bovine IFNγ capture mAb or IL-17A capture mAb (both from Kingfisher Biotech, Inc., St. Paul, MN), followed by a blocking step in cRPMI, for 2 h at 37 °C, 5% CO 2 .

Techniques: Control, Isolation, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Characterization of local and circulating bovine γδ T cell responses to respiratory BCG vaccination

doi: 10.1038/s41598-019-52565-z

Figure Lengend Snippet: Long-term cultured and ex vivo IFNγ responses by cattle after M. bovis aerosol vaccination. Cultured ELISPOT analysis was performed ~8 weeks after BCG aerosol vaccination. ( A ) For the ex vivo response, freshly isolated PBMCs were stimulated with rTb10.4/Ag85a or PPD-b or medium alone for 20 h (IFNγ) or for 48 hours (IL-17A). (B) Long-term cultured cells were generated by stimulating PBMC with a cocktail of rTB10.4/rAg85A (1 μg/ml each) and PPD-b (200 IU/ml) for 13 days followed by transfer to ELISPOT plates with APCs and the addition of either rTb10.4/Ag85a, PPD-b or medium alone. Medium control responses were subtracted from antigen-stimulated responses and results are presented as mean spot forming cells (SFC)/10 6 cells ( ± SEM) for ( A ) ex vivo conditions or ( B) long-term culture *p < 0.05 **p < 0.01 ***p < 0.001 as determined by Student’s t test. This figure was previously published and is duplicated with permission.

Article Snippet: Briefly, 96-well ELISPOT plates (Millipore) were coated at 4 °C overnight with an anti-bovine IFNγ capture mAb or IL-17A capture mAb (both from Kingfisher Biotech, Inc., St. Paul, MN), followed by a blocking step in cRPMI, for 2 h at 37 °C, 5% CO 2 .

Techniques: Cell Culture, Ex Vivo, Aerosol, Enzyme-linked Immunospot, Isolation, Generated, Control

Journal: Acta pharmacologica Sinica

Article Title: Pirfenidone ameliorates silica-induced lung inflammation and fibrosis in mice by inhibiting the secretion of interleukin-17A.

doi: 10.1038/s41401-021-00706-4

Figure Lengend Snippet: Fig. 3 Pirfenidone inhibited the secretion of IL-17A in early-stage silicosis models. ELISA was used to detect the protein levels of IL-17A in serum (a) and BALF (b). The mRNA and protein levels of IL-17A in lung tissues from different groups were measured by qPCR (c), immunohistochemistry (d), and Western blot analysis (e). All data were presented as mean ± SD; n = 10 for each group; ***P < 0.001, ****P < 0.0001, ns (no statistical significance).

Article Snippet: IL-17A neutralization By intraperitoneal injection, 100 μg of anti-mouse IL-17A antibody (M00421-2, BOSTER, Wuhan, China) or isotype control IgG (ab18443, abcam, Cambridge, UK) was given to silica-exposed mice weekly for 4 weeks, starting 1 day after silica exposure.

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

Journal: Acta pharmacologica Sinica

Article Title: Pirfenidone ameliorates silica-induced lung inflammation and fibrosis in mice by inhibiting the secretion of interleukin-17A.

doi: 10.1038/s41401-021-00706-4

Figure Lengend Snippet: Fig. 6 Pirfenidone diminished the production of IL-17A in advanced silicosis models. ELISA was performed to measure the protein expression of IL-17A in serum (a) and BALF (b) from different groups. The mRNA and protein levels of IL-17A in lung tissues were measured by qPCR (c), immunohistochemistry (d) and Western blot analysis (e), respectively. All data were presented as mean ± SD; n = 10 for each group; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: IL-17A neutralization By intraperitoneal injection, 100 μg of anti-mouse IL-17A antibody (M00421-2, BOSTER, Wuhan, China) or isotype control IgG (ab18443, abcam, Cambridge, UK) was given to silica-exposed mice weekly for 4 weeks, starting 1 day after silica exposure.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Immunohistochemistry, Western Blot

Journal: Annals of Dermatology

Article Title: Hyaluronan Oligosaccharides Improve Rosacea-Like Phenotype through Anti-Inflammatory and Epidermal Barrier-Improving Effects

doi: 10.5021/ad.2020.32.3.189

Figure Lengend Snippet: Treatment with hyaluronan oligosaccharides (oligo-HAs) reduced inflammatory cell infiltration, interleukin-17 (IL-17), and kallikrein 5 (KLK5) expressions. (A) Hematoxylin and eosin-stained tissue sections of LL-37 induced rosacea-like mice showed marked inflammatory cellular infiltration. Further treatments with oligo-HAs reduced cellular infiltration (H&E, ×100). (B) Majority of infiltrated cells were CD4+ T cells and infiltration of CD4+ T cells decreased in oligo-HAs treated group (CD4, ×100). (C) IL-17A expression was markedly increased in the epidermis of LL-37 induced rosacea-like mice. Oligo-HAs decreased the IL-17A expression regardless of the injection method (IL-17A, ×200). (D) Expression of KLK5 was increased in LL-37 induced rosacea-like mice (arrow head). Further treatment with oligo-HAs reduced KLK5 expression (KLK5, ×400). i.d.: intradermally, i.p.: intraperitoneally.

Article Snippet: To detect CD4, IL-17A, KLK5, filaggrin, and CD44 in histological specimens from mice, primary antibodies including a monoclonal mouse anti-CD4 antibody (BD Biosciences, Franklin Lakes, NJ, USA), monoclonal rat anti-mouse IL-17A antibody (Covalab, Villeurbanne, France), polyclonal rabbit anti-KLK5 antibody (Abcam, Cambridge, UK), polyclonal mouse anti-profilaggrin antibody (Covance, Princeton, NJ, USA), and monoclonal rat anti-human/mice CD44 antibody (eBioscience, San Diego, CA, USA) were used.

Techniques: Staining, Expressing, Injection