Huizhi Wang1, Mark W Graves2, Huaxin Zhou3, Zhen Gu4, Richard J Lamont5, David A Scott6. 1. Department of Oral Immunology and Infectious Diseases, University of Louisville, 501 South Preston Street, Louisville, KY, 40292, USA. huizhi.wang@louisville.edu. 2. Department of Oral Immunology and Infectious Diseases, University of Louisville, 501 South Preston Street, Louisville, KY, 40292, USA. mark.graves@louisville.edu. 3. Department of Oral Immunology and Infectious Diseases, University of Louisville, 501 South Preston Street, Louisville, KY, 40292, USA. h0zhou03@louisville.edu. 4. Department of Oral Immunology and Infectious Diseases, University of Louisville, 501 South Preston Street, Louisville, KY, 40292, USA. zhen.gu@louisville.edu. 5. Department of Oral Immunology and Infectious Diseases, University of Louisville, 501 South Preston Street, Louisville, KY, 40292, USA. rich.lamont@louisville.edu. 6. Department of Oral Immunology and Infectious Diseases, University of Louisville, 501 South Preston Street, Louisville, KY, 40292, USA. dascot07@louisville.edu.
Abstract
OBJECTIVE AND DESIGN: To elucidate the influence of 2-amino-4-(3,4-(methylenedioxy)benzylamino)-6-(3-methoxyphenyl)pyrimidine (AMBMP), a canonical Wnt/β-catenin pathway activator, on the inflammatory response of TLR-engaged innate cells in vitro. MATERIAL OR SUBJECT: Primary human monocytes. TREATMENT: AMPMB (0-10 μM), LPS (0-1.0 μg/ml), Pam3CSK4, FSL-1, or S. typhimurium flagellin (0-0.25 μg/ml). METHODS: TLR-induced cytokine release (TNF, IL-6, IL-12 p40) was monitored by ELISA while Wnt-related signals (GSK3β, p65, IκB, β-catenin) were assessed by Western blot, pharmaceutical inhibition and gene silencing. RESULTS: AMBMP induced the rapid phosphorylation of NFκB p65 at Ser(536) and abrogated total IκB, accompanied by a subsequent increase in pro-inflammatory cytokine production (TNF, IL-6, IL-12 p40) in otherwise naive monocytes. However, in TLR2, -4 and -5-engaged monocytes, AMBMP-suppressed cytokine production. In the context of LPS stimulation, this occurred concomitant with the phosphorylative inactivation of GSK3β at Ser(9), β-catenin accumulation and abrogation of NFκB p65 phosphorylation. AMBMP-mediated suppression of the TLR4 -induced inflammatory response was reversed by two pharmaceutical Wnt/β-catenin pathway inhibitors, IWP-2 and PNU-74654 and by Wnt3a silencing. CONCLUSIONS: Herein, we show that AMBMP induces canonical Wnt signaling events and acts as a suppressor of inflammation in surface TLR-engaged primary human monocytes.
OBJECTIVE AND DESIGN: To elucidate the influence of 2-amino-4-(3,4-(methylenedioxy)benzylamino)-6-(3-methoxyphenyl)pyrimidine (AMBMP), a canonical Wnt/β-catenin pathway activator, on the inflammatory response of TLR-engaged innate cells in vitro. MATERIAL OR SUBJECT: Primary human monocytes. TREATMENT: AMPMB (0-10 μM), LPS (0-1.0 μg/ml), Pam3CSK4, FSL-1, or S. typhimurium flagellin (0-0.25 μg/ml). METHODS: TLR-induced cytokine release (TNF, IL-6, IL-12 p40) was monitored by ELISA while Wnt-related signals (GSK3β, p65, IκB, β-catenin) were assessed by Western blot, pharmaceutical inhibition and gene silencing. RESULTS:AMBMP induced the rapid phosphorylation of NFκB p65 at Ser(536) and abrogated total IκB, accompanied by a subsequent increase in pro-inflammatory cytokine production (TNF, IL-6, IL-12 p40) in otherwise naive monocytes. However, in TLR2, -4 and -5-engaged monocytes, AMBMP-suppressed cytokine production. In the context of LPS stimulation, this occurred concomitant with the phosphorylative inactivation of GSK3β at Ser(9), β-catenin accumulation and abrogation of NFκB p65 phosphorylation. AMBMP-mediated suppression of the TLR4 -induced inflammatory response was reversed by two pharmaceutical Wnt/β-catenin pathway inhibitors, IWP-2 and PNU-74654 and by Wnt3a silencing. CONCLUSIONS: Herein, we show that AMBMP induces canonical Wnt signaling events and acts as a suppressor of inflammation in surface TLR-engaged primary human monocytes.
Authors: Michael Kuncewitch; Weng-Lang Yang; Asha Jacob; Adam Khader; Matthew Giangola; Jeffrey Nicastro; Gene F Coppa; Ping Wang Journal: J Trauma Acute Care Surg Date: 2015-04 Impact factor: 3.313
Authors: Jun Wei; Feng Fang; Anna P Lam; Jennifer L Sargent; Emily Hamburg; Monique E Hinchcliff; Cara J Gottardi; Radhika Atit; Michael L Whitfield; John Varga Journal: Arthritis Rheum Date: 2012-08
Authors: Rosa Di Liddo; Thomas Bertalot; Anne Schuster; Sandra Schrenk; Alessia Tasso; Ilenia Zanusso; Maria Teresa Conconi; Karl Herbert Schäfer Journal: J Neuroinflammation Date: 2015-02-03 Impact factor: 8.322