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BAFF Antagonist Attenuates the Development of Skin Fibrosis in Tight-Skin Mice  Takashi Matsushita, Manabu Fujimoto, Minoru Hasegawa, Yukiyo Matsushita,

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Presentation on theme: "BAFF Antagonist Attenuates the Development of Skin Fibrosis in Tight-Skin Mice  Takashi Matsushita, Manabu Fujimoto, Minoru Hasegawa, Yukiyo Matsushita,"— Presentation transcript:

1 BAFF Antagonist Attenuates the Development of Skin Fibrosis in Tight-Skin Mice 
Takashi Matsushita, Manabu Fujimoto, Minoru Hasegawa, Yukiyo Matsushita, Kazuhiro Komura, Fumihide Ogawa, Rei Watanabe, Kazuhiko Takehara, Shinichi Sato  Journal of Investigative Dermatology  Volume 127, Issue 12, Pages (December 2007) DOI: /sj.jid Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 BAFF is increased in sera from TSK/+ and NZB/NZW F1 mice. (a) Serum BAFF levels were determined by ELISA. Horizontal bars represent mean serum BAFF level in each group. *P<0.01, **P<0.05 versus wild type. (b) Representative expression of BAFF-R, including BAFF-receptor, TACI, and BCMA, on B220+ B cells from 8-week-old TSK/+ and wild-type littermates. Three BAFF-receptor expressions were assessed by two-color immunofluorescence with flow cytometric analysis. Shaded regions in the histograms represent TSK/+ B220+ B cells, boldface lines represent wild-type B220+ B cells, and dotted lines represent isotype control staining. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 Effect of BAFF antagonist on skin fibrosis in TSK/+ mice. Skin fibrosis in dorsal skin from 8-week-old TSK/+ and wild-type littermates treated with BAFF-R-Ig or Fc control protein (control). (a) Skin fibrosis was assessed by quantitatively measuring hypodermal thickness. Representative hematoxylin and eosin-stained histological sections. Dermis is indicated by (d), hypodermis beneath the panniculus carnosus (arrow) indicated by (h). Bar=0.1mm. These results represent those obtained with at least five mice of each group. Skin fibrosis was assessed further by quantitatively measuring (b) hypodermal thickness and (c) skin collagen content. Results from each mouse are represented as single dots. Horizontal bars represent mean hypodermal thickness or collagen content in each group. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 BAFF antagonist modifies B-cell phenotypes in TSK/+ mice. B-cell phenotypes in 8-week-old TSK/+ and wild-type littermates treated with BAFF-R-Ig or Fc control protein (control). Bone marrow and spleen were collected, stained with fluorescently labeled mAbs to a variety of cell surface markers, and analyzed by flow cytometry. Representative profiles are shown for bone marrow and B220+ spleen. (a) Bone marrow cells were stained with mAbs to IgM and B220, to identify pro/pre (IgM−B220low), immature (IgM+B220low), and mature B cells (IgM+B220high). (b) Splenic B cells (B220+-gated) were stained with mAbs, CD21, CD23, and CD24, to identify newly formed (NF; CD21−CD23−B220+), T1 (CD24highCD21− B220+), T2 (CD24highCD21+ B220+), follicular (FO; CD21+ CD23+ B220+), and marginal zone (MZ) B cells (CD21highCD23− B220+). Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 BAFF antagonist inhibits autoantibody generation and hyper-γ-globulinemia in TSK/+ mice. (a) Anti-topoisomerase I (topo I) Ab and (b) antibody levels in sera from 8-week-old TSK/+ and wild-type littermates treated with BAFF-R-Ig or Fc control protein (control). Relative serum autoantibody and antibody levels were determined by Ig subclass-specific ELISA. Horizontal bars represent mean autoantibody and antibody levels for each group. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 BAFF antagonist downregulates augmented type 2 cytokines in TSK/+ skin. Messenger RNA expression of cytokines in dorsal skin from 8-week-old TSK/+ and wild-type littermates treated with BAFF-R-Ig or Fc control protein (control). The mRNA levels of TGF-β1, IL-4, IL-6, IL-10, and IFN-γ were analyzed by real-time reverse transcriptase-PCR, and normalized with the internal control glyceraldehyde-3-phosphate dehydrogenase. Each sample was done in triplicate. Data indicate the mean±SEM. These results represent those obtained with at least five mice of each group. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

7 Figure 6 BAFF accelerates IL-6 production by TSK/+ B cells. Splenic B cells were purified by positive selection with anti-B220 Ab-coated magnetic beads and were stimulated for 72hours with either medium alone or recombinant mouse BAFF and/or Staphylococcus aureus Cowan strain (SAC) and/or BAFF-R-Ig. Culture supernatants were analyzed by ELISA to determine the amount of IL-6 and IL-10. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

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