monoclonal antibodies against il 17  (Novartis)

Journal: PLoS ONE

Article Title: Healthy Human T-Cell Responses to Aspergillus fumigatus Antigens

doi: 10.1371/journal.pone.0009036

Figure Lengend Snippet: (a) IFN-γ (b) IL-4 and (c) IL-17 ELISPOT assay. Blood was collected from 20 healthy donors and triplicate samples of 1×10 5 (IFN-γ) or 2×10 5 PBMCs (IL-4 or IL-17) were stimulated with medium, 5 µg/ml phytohemoagglutinin (PHA) or 1 µg/ml CHE or recombinant A. fumigatus antigens for 24–48 hrs at 37°C/5%CO 2 . The mean antigen-specific spot forming cells, SFC (after background subtraction of control wells with no antigen) is plotted. The median for each set of healthy individuals is indicated by a bar. The figures show one representative data from three independent experiments. *** P <0.001, ** P <0.01 versus CHE stimulated PBMC.

Article Snippet: Briefly, ELISPOT plates (Multiscreen-MAIPS4510, Millipore) were coated using primary monoclonal antibodies against IFN-γ (Clone 1-D1K, MabTech, Sweden), IL-4 (82.4, MabTech, Sweden) and IL-17 (clone eBio64CAP17, eBioscience, San Diego, CA) at the concentration of 10 µg/ml in bicarbonate coating buffer, pH-9.6.

Techniques: Enzyme-linked Immunospot, Recombinant

Journal: PLoS ONE

Article Title: Healthy Human T-Cell Responses to Aspergillus fumigatus Antigens

doi: 10.1371/journal.pone.0009036

Figure Lengend Snippet: (a) IFN-γ (b) IL-4 and (c) IL-17 responses. Blood was collected from 20 healthy donors and triplicate samples of 1×10 5 PBMCs were stimulated with medium, 5 µg/ml PHA or 1 µg/ml CHE or recombinant A. fumigatus antigens for 24–48 hr at 37°C/5%CO 2 . The mean antigen-specific spot forming cells, SFC (after background subtraction of control wells with no antigen) is plotted.

Article Snippet: Briefly, ELISPOT plates (Multiscreen-MAIPS4510, Millipore) were coated using primary monoclonal antibodies against IFN-γ (Clone 1-D1K, MabTech, Sweden), IL-4 (82.4, MabTech, Sweden) and IL-17 (clone eBio64CAP17, eBioscience, San Diego, CA) at the concentration of 10 µg/ml in bicarbonate coating buffer, pH-9.6.

Techniques: Recombinant

Journal: iScience

Article Title: SPARC regulation of PMN clearance protects from pristane-induced lupus and rheumatoid arthritis

doi: 10.1016/j.isci.2021.102510

Figure Lengend Snippet: Increased IL-17 production in arthritic paws from Sparc −/− mice (A−L) (A) Intracellular staining for IL-17, IFNγ, and TNF in CD4 + T cells from draining LN of naive and pristane-treated mice (130 days after treatment). Results are representative of three independent experiments (∗p < 0.05, Student's t test; error bars are mean ± SD). (B) Representative dot plots for the intracellular staining. (C) qPCR for IL-17A performed by digesting the whole paws of the indicated mice; results are normalized to GAPDH; representative of two other independent experiments; (∗p < 0.05, Student's t test; error bars are mean ± SD). (D) Intracellular staining for IL-17 in Th cells (CD45 + CD4 + ), PMN (CD45 + CD11b + Gr1 hi ), and other infiltrating cells (CD45 + CD11b − CD4 - ) of paws isolated and digested from mice 130 days after pristane injection. (∗p < 0.05, Student's t test; error bars are mean ± SD). (E) Production of IL-17 by γδTCR and CD4 cells. Representative plots. (F) IHC analysis for Gr-1 performed on paws obtained from WT and Sparc −/− mice. Scale bar, 500 μm. (G) Frequency of proliferating γδT cells when co-cultured with DC and PMN either from WT or Sparc −/− mice. This analysis highlights a direct capacity of Sparc −/− PMN (either naive or undergoing NETosis) of stimulating γδT cells, independent of the presence of DCs (∗p < 0.05; ∗∗p < 0.01, Student's t test, mean ± SEM). (H) Arthritis score obtained evaluating arthritis-inducing antibody cocktail (ArthritoMab)-treated mice every 2 weeks (n = 17 mice/group pooled from two independent experiments, ∗p< 0.05) (I) Representative pictures of paws from WT and Sparc −/− at day 200 treated with ArthritoMab. (J) Frequency of PMN (mean ± SEM), (K) γδT cells (mean ± SEM), and (L) IL-17+ γδT cells (mean ± SEM) in mice treated with ArthritoMab (∗p < 0.05; ∗∗∗p < 0.001; Student's t test).

Article Snippet: Then cells were surface stained as above and fixed 20 min using the IC fixation buffer, followed by incubation with monoclonal antibodies against IL-17, IL-4, TNF, IFNγ, and resuspendend in Permeabilization Buffer (all from eBioscience).

Techniques: Staining, Isolation, Injection, Cell Culture