BACKGROUND AND PURPOSE: A series of benzothiazole derivatives were screened for immunosuppressive activity; of these compounds BD750 was found to be the most effective immunosuppressant. The purpose of the current study was to determine the immunosuppressive activity of BD750 on T cell proliferation and its potential mode of action. EXPERIMENTAL APPROACH: T cell proliferation, CD25 and CD69 expression and cell cycle distribution were measured in vitro by flow cytometry. Cell viability was determined by CCK-8 assay. Cytokine levels were measured by elisa. The activation of signal-regulated molecules was assessed by Western blot analysis. The effects of BD750 were evaluated in vivo in a mouse model of delayed-type hypersensitivity. KEY RESULTS: BD750 significantly inhibited mouse and human T cell proliferation, stimulated either by anti-CD3/anti-CD28 monoclonal antibodies or by an alloantigen, in a dose-dependent manner in vitro. No obvious cytotoxic effects of BD750 were observed in our experimental conditions. Furthermore, BD750 did not inhibit CD25 and CD69 expression or IL-2 and IL-4 secretion, but induced cell cycle arrest at the G(0) /G(1) phase in activated T cells. In IL-2-stimulated CTLL-2 cells and primary activated T cells, BD750 inhibited cell proliferation and STAT5 phosphorylation, but not Akt or p70S6K phosphorylation. BD750 also reduced the T cell-mediated delayed-type hypersensitivity response in mice in a dose-dependent manner. CONCLUSION AND IMPLICATIONS: These data indicate that BD750 inhibits IL-2-induced JAK3/STAT5-dependent T cell proliferation. BD750 has the potential to be used as a lead compound for the design and development of new immunosuppressants for preventing graft rejection and treating autoimmune diseases.
BACKGROUND AND PURPOSE: A series of benzothiazole derivatives were screened for immunosuppressive activity; of these compounds BD750 was found to be the most effective immunosuppressant. The purpose of the current study was to determine the immunosuppressive activity of BD750 on T cell proliferation and its potential mode of action. EXPERIMENTAL APPROACH: T cell proliferation, CD25 and CD69 expression and cell cycle distribution were measured in vitro by flow cytometry. Cell viability was determined by CCK-8 assay. Cytokine levels were measured by elisa. The activation of signal-regulated molecules was assessed by Western blot analysis. The effects of BD750 were evaluated in vivo in a mouse model of delayed-type hypersensitivity. KEY RESULTS:BD750 significantly inhibited mouse and human T cell proliferation, stimulated either by anti-CD3/anti-CD28 monoclonal antibodies or by an alloantigen, in a dose-dependent manner in vitro. No obvious cytotoxic effects of BD750 were observed in our experimental conditions. Furthermore, BD750 did not inhibit CD25 and CD69 expression or IL-2 and IL-4 secretion, but induced cell cycle arrest at the G(0) /G(1) phase in activated T cells. In IL-2-stimulated CTLL-2 cells and primary activated T cells, BD750 inhibited cell proliferation and STAT5 phosphorylation, but not Akt or p70S6K phosphorylation. BD750 also reduced the T cell-mediated delayed-type hypersensitivity response in mice in a dose-dependent manner. CONCLUSION AND IMPLICATIONS: These data indicate that BD750 inhibits IL-2-induced JAK3/STAT5-dependent T cell proliferation. BD750 has the potential to be used as a lead compound for the design and development of new immunosuppressants for preventing graft rejection and treating autoimmune diseases.
Authors: Mark E Flanagan; Todd A Blumenkopf; William H Brissette; Matthew F Brown; Jeffrey M Casavant; Chang Shang-Poa; Jonathan L Doty; Eileen A Elliott; Michael B Fisher; Michael Hines; Craig Kent; Elizabeth M Kudlacz; Brett M Lillie; Kelly S Magnuson; Sandra P McCurdy; Michael J Munchhof; Bret D Perry; Perry S Sawyer; Timothy J Strelevitz; Chakrapani Subramanyam; Jianmin Sun; David A Whipple; Paul S Changelian Journal: J Med Chem Date: 2010-11-24 Impact factor: 7.446
Authors: H Takenaka; S Maruo; N Yamamoto; M Wysocka; S Ono; M Kobayashi; H Yagita; K Okumura; T Hamaoka; G Trinchieri; H Fujiwara Journal: J Leukoc Biol Date: 1997-01 Impact factor: 4.962
Authors: Paul S Changelian; Mark E Flanagan; Douglas J Ball; Craig R Kent; Kelly S Magnuson; William H Martin; Bonnie J Rizzuti; Perry S Sawyer; Bret D Perry; William H Brissette; Sandra P McCurdy; Elizabeth M Kudlacz; Maryrose J Conklyn; Eileen A Elliott; Erika R Koslov; Michael B Fisher; Timothy J Strelevitz; Kwansik Yoon; David A Whipple; Jianmin Sun; Michael J Munchhof; John L Doty; Jeffrey M Casavant; Todd A Blumenkopf; Michael Hines; Matthew F Brown; Brett M Lillie; Chakrapani Subramanyam; Chang Shang-Poa; Anthony J Milici; Gretchen E Beckius; James D Moyer; Chunyan Su; Thasia G Woodworth; Anderson S Gaweco; Chan R Beals; Bruce H Littman; Douglas A Fisher; James F Smith; Panayiotis Zagouras; Holly A Magna; Mary J Saltarelli; Kimberly S Johnson; Linda F Nelms; Shelley G Des Etages; Lisa S Hayes; Thomas T Kawabata; Deborah Finco-Kent; Deanna L Baker; Michael Larson; Ming-Sing Si; Ricardo Paniagua; John Higgins; Bari Holm; Bruce Reitz; Yong-Jie Zhou; Randall E Morris; John J O'Shea; Dominic C Borie Journal: Science Date: 2003-10-31 Impact factor: 47.728