bovine recombinant il 17a  (Kingfisher Biotech)

Journal: Veterinary Research

Article Title: Comparison of innate immune agonists for induction of tracheal antimicrobial peptide gene expression in tracheal epithelial cells of cattle

doi: 10.1186/s13567-014-0105-8

Figure Lengend Snippet: Effects of CpG oligodinucleotide, interleukin-17A and interferon-α on tracheal antimicrobial peptide gene expression. Dose- and time-dependent effects were measured for CpG oligodinucleotide (A,B) , a TLR9 agonist; interleukin-17A (C,D) ; and interferon-α (E,F) . For the dose–response studies (A,C,E) , confluent bTEC were stimulated in triplicate with various concentrations of agonist for 16 h. For the time-course studies (B,D,F) , confluent bTEC were stimulated in triplicate for 4, 8, 16 or 24 h with the doses of agonist shown. Gene expression of TAP relative to that of GAPDH was measured using real-time RT-qPCR. Lipopolysaccharide (LPS, 0.1 μg/mL) was used in all assays as a positive control and standard. *, significantly different from unstimulated cells ( P

Article Snippet: The cytokines included bovine recombinant IL-17A (Kingfisher Biotech, St. Paul, MN, USA, item number RP0056B-005) and bovine recombinant IFN-α (Kingfisher Biotech, item RP0008B-025).

Techniques: Expressing, Quantitative RT-PCR, Positive Control

Journal: Veterinary Research

Article Title: Comparison of innate immune agonists for induction of tracheal antimicrobial peptide gene expression in tracheal epithelial cells of cattle

doi: 10.1186/s13567-014-0105-8

Figure Lengend Snippet: Comparison of the effects of IL-17A, Pam3CSK4 and LPS on tracheal antimicrobial peptide gene expression. Confluent cultures of tracheal epithelial cells from 4 different calves were non-stimulated (NS) or stimulated with 1 μg/mL Pam3CSK4, 316 ng/mL IL-17A, or 0.1 μg/mL LPS for 8 h (A) and 16 h (B) in triplicate. Gene expression was assessed using real-time RT-qPCR. Pam3CSK4 induced significantly higher tracheal antimicrobial peptide gene expression than IL-17A and LPS at both 8 and 16 h ( P

Article Snippet: The cytokines included bovine recombinant IL-17A (Kingfisher Biotech, St. Paul, MN, USA, item number RP0056B-005) and bovine recombinant IFN-α (Kingfisher Biotech, item RP0008B-025).

Techniques: Expressing, Quantitative RT-PCR

Journal: Veterinary Research

Article Title: Comparison of innate immune agonists for induction of tracheal antimicrobial peptide gene expression in tracheal epithelial cells of cattle

doi: 10.1186/s13567-014-0105-8

Figure Lengend Snippet: Effect of stimulation with single agonists compared to combined agonists. Cultured bovine tracheal epithelial cells were stimulated for 16 h in triplicate with various combinations of 1 μg/mL Pam3CSK4, 316 ng/mL IL-17A, or 0.1 μg/mL LPS. Tracheal antimicrobial peptide gene expression was measured as above. The effects of combined agonists were greater than that of interleukin-17A (IL-17A) alone, but minimally or not different than that of lipopolysaccharide (LPS) or Pam3CSK4 alone. The data shown (panels A, B and C) represent 3 studies conducted on different days using cells from different calves.

Article Snippet: The cytokines included bovine recombinant IL-17A (Kingfisher Biotech, St. Paul, MN, USA, item number RP0056B-005) and bovine recombinant IFN-α (Kingfisher Biotech, item RP0008B-025).

Techniques: Cell Culture, Expressing

Journal: Veterinary Research

Article Title: Regulation of tracheal antimicrobial peptide gene expression in airway epithelial cells of cattle

doi: 10.1186/s13567-016-0329-x

Figure Lengend Snippet: Effect of dexamethasone on LPS-, Pam3CSK4- and IL-17A-induced up-regulation of TAP mRNA. Bovine tracheal epithelial cells were grown in the presence or absence of 10 −6 M dexamethasone for 24 h. LPS (0.1 μg/mL), Pam3CSK4 (1 μg/mL) or IL-17A (316 ng/mL) were added for 16 h. Each of these agonists significantly upregulated TAP gene expression relative to the reference gene GAPDH ( P

Article Snippet: Triplicate cell cultures supplemented with DMEM/F12 without serum were unstimulated, or stimulated with 0.1 μg/mL LPS (Sigma Aldrich, MO, USA, L9143), 1 μg/mL Pam3CSK4 (Invivogen, San Diego, CA, USA, t1rl-pms), or 316 ng/mL IL-17A (Kingfisher Biotech, St. Paul, MN, USA, RP0056B-005) for 16 h. The doses and time were optimized in a previous study [ ].

Techniques: Expressing

Journal: Veterinary Research

Article Title: Regulation of tracheal antimicrobial peptide gene expression in airway epithelial cells of cattle

doi: 10.1186/s13567-016-0329-x

Figure Lengend Snippet: Effect of the NF-κB inhibitor caffeic acid phenylester (CAPE) on induction of TAP gene expression by LPS, Pam3CSK4 and IL-17A. Stimulation of bovine tracheal epithelial cells (bTEC) with 0.1 µg/mL LPS (panel A ), 1 µg/mL Pam3CSK4 (panel B ), or 316 ng/mL IL-17A (panel C ) induced significantly greater TAP gene expression than in non-stimulated cells (NS). Pre-treatment with CAPE for 2 h before stimulation inhibited each of these responses. Asterisks indicate significant effects of CAPE treatment compared to no CAPE, P

Article Snippet: Triplicate cell cultures supplemented with DMEM/F12 without serum were unstimulated, or stimulated with 0.1 μg/mL LPS (Sigma Aldrich, MO, USA, L9143), 1 μg/mL Pam3CSK4 (Invivogen, San Diego, CA, USA, t1rl-pms), or 316 ng/mL IL-17A (Kingfisher Biotech, St. Paul, MN, USA, RP0056B-005) for 16 h. The doses and time were optimized in a previous study [ ].

Techniques: Expressing

Journal: Veterinary Research

Article Title: Regulation of tracheal antimicrobial peptide gene expression in airway epithelial cells of cattle

doi: 10.1186/s13567-016-0329-x

Figure Lengend Snippet: Western analysis of NF-kB p65 in nuclear and cytoplasmic extracts of bTEC stimulated with LPS, Pam3CSK4 or IL-17A. NF-kB p65 was localized to the cytoplasm of non-stimulated cells (NS), but showed translocation to the nucleus following treatment of the cells with either LPS (panel A ), Pam3CSK4 (Pam, panel B ) or IL-17A (panel C ). However, pre-treating the cells with the NF-κB inhibitor CAPE (10 μM) prior to exposure to agonists fully abrogated nuclear translocation of NF-kB p65 in the studies involving LPS and Pam3CSK4 (panels A and B ), and partially in the studies involving IL-17A (panel C ). Pos control: positive control, NF-kB p65. The findings were similar when the experiments were repeated using cells from different animals.

Article Snippet: Triplicate cell cultures supplemented with DMEM/F12 without serum were unstimulated, or stimulated with 0.1 μg/mL LPS (Sigma Aldrich, MO, USA, L9143), 1 μg/mL Pam3CSK4 (Invivogen, San Diego, CA, USA, t1rl-pms), or 316 ng/mL IL-17A (Kingfisher Biotech, St. Paul, MN, USA, RP0056B-005) for 16 h. The doses and time were optimized in a previous study [ ].

Techniques: Western Blot, Translocation Assay, Positive Control