Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-17B and IL-17C are associated with TNF-alpha production and contribute to the exacerbation of inflammatory arthritis.

doi: 10.4049/jimmunol.179.10.7128

Figure Lengend Snippet: FIGURE 1. The expression of IL-17 family members and IL-17R genes in the arthritic paws of CIA mice. A, The expressions of IL-17 family genes and IL-17R genes were examined in the arthritic paws of CIA mice (f; n 3) and in control mice (; n 3) by quantitative PCR. B, The expressions of inflammatory cy- tokines. C, The expressions of IL-17 family members in the sorted cell populations of the arthritic paws of CIA mice. The data are representative of three independent experiments.

Article Snippet: These cells were stained with anti-mouse IL-17A mAb conjugated to PE (BD Pharmingen), biotinylated anti-mouse IL-17B polyclonal Ab (R&D Systems), anti-mouse IL-17C polyclonal Ab (R&D Systems), and antimouse IL-17F mAb (R&D Systems), respectively.

Techniques: Expressing, Control, Real-time Polymerase Chain Reaction

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-17B and IL-17C are associated with TNF-alpha production and contribute to the exacerbation of inflammatory arthritis.

doi: 10.4049/jimmunol.179.10.7128

Figure Lengend Snippet: FIGURE 2. The proinflammatory effects of IL-17 family members on mouse fibroblasts and macrophages. A, Relative expression of the cytokine genes in 3T3 cell. The mouse fibroblast cell line 3T3 was cultured with each of mIL-17A, mIL-17B, mIL-17C, or mIL-17F for 24 h, and the expressions of inflammatory cytokines were measured by quantitative PCR. B, Relative expression of the cytokine genes in mouse thioglycolate-elicited PECs. PECs were cultured with each of mIL-17A, mIL-17B, mIL-17C, or mIL-17F for 24 h, and the expressions of inflammatory cytokines were measured by quantitative PCR. C, The secreted IL-6 and TNF- levels in the supernatants of 3T3 and PECs were measured by ELISA. Error bars indicate SD. The data are representative of three independent experiments. Significance of differences between control (medium) and each IL-17 family was determined; , p 0.05.

Article Snippet: These cells were stained with anti-mouse IL-17A mAb conjugated to PE (BD Pharmingen), biotinylated anti-mouse IL-17B polyclonal Ab (R&D Systems), anti-mouse IL-17C polyclonal Ab (R&D Systems), and antimouse IL-17F mAb (R&D Systems), respectively.

Techniques: Expressing, Cell Culture, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-17B and IL-17C are associated with TNF-alpha production and contribute to the exacerbation of inflammatory arthritis.

doi: 10.4049/jimmunol.179.10.7128

Figure Lengend Snippet: FIGURE 3. The effects of transfer of IL-17 family-transduced CD4 T cells on CIA. A, Intracellular IL-17 family expressions in Ba/F3 cells ret- rovirally transduced with each IL-17 family member. GFP-gated IL-17 family-transduced (mIL-17A, mIL- 17B, mIL-17C, or mIL-17F) Ba/F3 cells were analyzed for IL-17A, IL- 17B, IL-17C, or IL-17F expression compared with GFP-gated empty vector (pMIG)-transduced Ba/F3 cells. B, Representative FACS analy- sis of IL-17 family-transduced CD4

Article Snippet: These cells were stained with anti-mouse IL-17A mAb conjugated to PE (BD Pharmingen), biotinylated anti-mouse IL-17B polyclonal Ab (R&D Systems), anti-mouse IL-17C polyclonal Ab (R&D Systems), and antimouse IL-17F mAb (R&D Systems), respectively.

Techniques: Transduction, Expressing, Plasmid Preparation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-17B and IL-17C are associated with TNF-alpha production and contribute to the exacerbation of inflammatory arthritis.

doi: 10.4049/jimmunol.179.10.7128

Figure Lengend Snippet: FIGURE 4. Generation of IL-17 family chimeric mice by BM trans- plantation of gene-transduced BM cells. Each of IL-17 family genes was transduced to BM cells with retrovirus vector and transferred to lethally irradiated mice. A, The intracellular expression of IL-17A protein in the spleen of IL-17A BM chimeric mice 8 wk after BM transplantation. The percentage of GFP cells expressing IL-17A is indicated. The data are representative of three independent experiments. B, The concentration of IL-17A protein in the serum of IL-17A BM chimeric mice (n 6) and control mice (pMIG BM chimeric mice) (n 6). The levels of IL-17A were measured by ELISA.

Article Snippet: These cells were stained with anti-mouse IL-17A mAb conjugated to PE (BD Pharmingen), biotinylated anti-mouse IL-17B polyclonal Ab (R&D Systems), anti-mouse IL-17C polyclonal Ab (R&D Systems), and antimouse IL-17F mAb (R&D Systems), respectively.

Techniques: Plasmid Preparation, Irradiation, Expressing, Transplantation Assay, Concentration Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-17B and IL-17C are associated with TNF-alpha production and contribute to the exacerbation of inflammatory arthritis.

doi: 10.4049/jimmunol.179.10.7128

Figure Lengend Snippet: FIGURE 5. Incidence of CIA and arthritis scores in IL-17 family BM chimeric mice. Incidence of CIA and arthritis scores in IL-17A and IL-17F BM chimeric mice (A and B), and in IL-17B and IL-17C BM chimeric mice (C and D). Mice were immu- nized with BCII 8 wk after the BM transplantation. Values are the mean of experiments for IL-17A and IL-17F BM chimeric mice (n 20 per group) and experiments for IL-17B and IL-17C BM chimeric mice (n 30 per group). Significance of differences between control (pMIG) and each IL-17 family BM chimeric mice was determined; , p 0.005; , p 0.05. E, The mRNA expression of inflammatory cytokines in the spleen of BM chi- meric mice of IL-17B and IL-17C, which were immunized with BCII (f; n 15 per group) or nonimmunized controls (; n 6 per group). Sig- nificance of differences between con- trol (pMIG) and each IL-17 family BM chimeric mice was determined; , p 0.005; , p 0.05. F, The secreted TNF- and IL-6 levels in the serum of IL-17 family BM chimeric mice that were immunized with BCII (f; n 15) or nonimmunized con- trols (; n 6). Significance of dif- ferences between control (pMIG) and each IL-17 family BM chimeric mice was determined; , p 0.005; , p 0.05.

Article Snippet: These cells were stained with anti-mouse IL-17A mAb conjugated to PE (BD Pharmingen), biotinylated anti-mouse IL-17B polyclonal Ab (R&D Systems), anti-mouse IL-17C polyclonal Ab (R&D Systems), and antimouse IL-17F mAb (R&D Systems), respectively.

Techniques: Transplantation Assay, Control, Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-17B and IL-17C are associated with TNF-alpha production and contribute to the exacerbation of inflammatory arthritis.

doi: 10.4049/jimmunol.179.10.7128

Figure Lengend Snippet: FIGURE 6. Effect of anti-IL-17B Ab treatment in CIA mice. A, CIA mice received i.p. injection of anti-mouse IL-17B Abs after the first clinical signs of arthritis (arthritis score between 1 and 2). As a control, PBS was injected. The arthritis score was shown. B, Histological score of the inflammatory joints of CIA mice treated with anti-IL-17B Abs was evaluated at 10 days after the onset of arthritis. Cellular infiltration and pannus invasion were graded in all four paws of the mice. Values are the mean of arthritis scores for anti-IL-17B Ab-treated mice and control mice (n 5 per group). Significance of differences between control and anti-IL-17B Ab-treated mice was shown.

Article Snippet: These cells were stained with anti-mouse IL-17A mAb conjugated to PE (BD Pharmingen), biotinylated anti-mouse IL-17B polyclonal Ab (R&D Systems), anti-mouse IL-17C polyclonal Ab (R&D Systems), and antimouse IL-17F mAb (R&D Systems), respectively.

Techniques: Injection, Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-17B and IL-17C are associated with TNF-alpha production and contribute to the exacerbation of inflammatory arthritis.

doi: 10.4049/jimmunol.179.10.7128

Figure Lengend Snippet: FIGURE 1. The expression of IL-17 family members and IL-17R genes in the arthritic paws of CIA mice. A, The expressions of IL-17 family genes and IL-17R genes were examined in the arthritic paws of CIA mice (f; n 3) and in control mice (; n 3) by quantitative PCR. B, The expressions of inflammatory cy- tokines. C, The expressions of IL-17 family members in the sorted cell populations of the arthritic paws of CIA mice. The data are representative of three independent experiments.

Article Snippet: Mice exhibited the first clinical signs of arthritis (arthritis score between 1 and 2) and were injected i.p. with 100 g of polyclonal anti-mouse IL-17B Abs (R&D Systems).

Techniques: Expressing, Control, Real-time Polymerase Chain Reaction

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-17B and IL-17C are associated with TNF-alpha production and contribute to the exacerbation of inflammatory arthritis.

doi: 10.4049/jimmunol.179.10.7128

Figure Lengend Snippet: FIGURE 2. The proinflammatory effects of IL-17 family members on mouse fibroblasts and macrophages. A, Relative expression of the cytokine genes in 3T3 cell. The mouse fibroblast cell line 3T3 was cultured with each of mIL-17A, mIL-17B, mIL-17C, or mIL-17F for 24 h, and the expressions of inflammatory cytokines were measured by quantitative PCR. B, Relative expression of the cytokine genes in mouse thioglycolate-elicited PECs. PECs were cultured with each of mIL-17A, mIL-17B, mIL-17C, or mIL-17F for 24 h, and the expressions of inflammatory cytokines were measured by quantitative PCR. C, The secreted IL-6 and TNF- levels in the supernatants of 3T3 and PECs were measured by ELISA. Error bars indicate SD. The data are representative of three independent experiments. Significance of differences between control (medium) and each IL-17 family was determined; , p 0.05.

Article Snippet: Mice exhibited the first clinical signs of arthritis (arthritis score between 1 and 2) and were injected i.p. with 100 g of polyclonal anti-mouse IL-17B Abs (R&D Systems).

Techniques: Expressing, Cell Culture, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-17B and IL-17C are associated with TNF-alpha production and contribute to the exacerbation of inflammatory arthritis.

doi: 10.4049/jimmunol.179.10.7128

Figure Lengend Snippet: FIGURE 3. The effects of transfer of IL-17 family-transduced CD4 T cells on CIA. A, Intracellular IL-17 family expressions in Ba/F3 cells ret- rovirally transduced with each IL-17 family member. GFP-gated IL-17 family-transduced (mIL-17A, mIL- 17B, mIL-17C, or mIL-17F) Ba/F3 cells were analyzed for IL-17A, IL- 17B, IL-17C, or IL-17F expression compared with GFP-gated empty vector (pMIG)-transduced Ba/F3 cells. B, Representative FACS analy- sis of IL-17 family-transduced CD4

Article Snippet: Mice exhibited the first clinical signs of arthritis (arthritis score between 1 and 2) and were injected i.p. with 100 g of polyclonal anti-mouse IL-17B Abs (R&D Systems).

Techniques: Transduction, Expressing, Plasmid Preparation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-17B and IL-17C are associated with TNF-alpha production and contribute to the exacerbation of inflammatory arthritis.

doi: 10.4049/jimmunol.179.10.7128

Figure Lengend Snippet: FIGURE 4. Generation of IL-17 family chimeric mice by BM trans- plantation of gene-transduced BM cells. Each of IL-17 family genes was transduced to BM cells with retrovirus vector and transferred to lethally irradiated mice. A, The intracellular expression of IL-17A protein in the spleen of IL-17A BM chimeric mice 8 wk after BM transplantation. The percentage of GFP cells expressing IL-17A is indicated. The data are representative of three independent experiments. B, The concentration of IL-17A protein in the serum of IL-17A BM chimeric mice (n 6) and control mice (pMIG BM chimeric mice) (n 6). The levels of IL-17A were measured by ELISA.

Article Snippet: Mice exhibited the first clinical signs of arthritis (arthritis score between 1 and 2) and were injected i.p. with 100 g of polyclonal anti-mouse IL-17B Abs (R&D Systems).

Techniques: Plasmid Preparation, Irradiation, Expressing, Transplantation Assay, Concentration Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-17B and IL-17C are associated with TNF-alpha production and contribute to the exacerbation of inflammatory arthritis.

doi: 10.4049/jimmunol.179.10.7128

Figure Lengend Snippet: FIGURE 5. Incidence of CIA and arthritis scores in IL-17 family BM chimeric mice. Incidence of CIA and arthritis scores in IL-17A and IL-17F BM chimeric mice (A and B), and in IL-17B and IL-17C BM chimeric mice (C and D). Mice were immu- nized with BCII 8 wk after the BM transplantation. Values are the mean of experiments for IL-17A and IL-17F BM chimeric mice (n 20 per group) and experiments for IL-17B and IL-17C BM chimeric mice (n 30 per group). Significance of differences between control (pMIG) and each IL-17 family BM chimeric mice was determined; , p 0.005; , p 0.05. E, The mRNA expression of inflammatory cytokines in the spleen of BM chi- meric mice of IL-17B and IL-17C, which were immunized with BCII (f; n 15 per group) or nonimmunized controls (; n 6 per group). Sig- nificance of differences between con- trol (pMIG) and each IL-17 family BM chimeric mice was determined; , p 0.005; , p 0.05. F, The secreted TNF- and IL-6 levels in the serum of IL-17 family BM chimeric mice that were immunized with BCII (f; n 15) or nonimmunized con- trols (; n 6). Significance of dif- ferences between control (pMIG) and each IL-17 family BM chimeric mice was determined; , p 0.005; , p 0.05.

Article Snippet: Mice exhibited the first clinical signs of arthritis (arthritis score between 1 and 2) and were injected i.p. with 100 g of polyclonal anti-mouse IL-17B Abs (R&D Systems).

Techniques: Transplantation Assay, Control, Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-17B and IL-17C are associated with TNF-alpha production and contribute to the exacerbation of inflammatory arthritis.

doi: 10.4049/jimmunol.179.10.7128

Figure Lengend Snippet: FIGURE 6. Effect of anti-IL-17B Ab treatment in CIA mice. A, CIA mice received i.p. injection of anti-mouse IL-17B Abs after the first clinical signs of arthritis (arthritis score between 1 and 2). As a control, PBS was injected. The arthritis score was shown. B, Histological score of the inflammatory joints of CIA mice treated with anti-IL-17B Abs was evaluated at 10 days after the onset of arthritis. Cellular infiltration and pannus invasion were graded in all four paws of the mice. Values are the mean of arthritis scores for anti-IL-17B Ab-treated mice and control mice (n 5 per group). Significance of differences between control and anti-IL-17B Ab-treated mice was shown.

Article Snippet: Mice exhibited the first clinical signs of arthritis (arthritis score between 1 and 2) and were injected i.p. with 100 g of polyclonal anti-mouse IL-17B Abs (R&D Systems).

Techniques: Injection, Control