Gp96 peptide antagonist gp96-II confers therapeutic effects in murine intestinal inflammation

Claudia A. Nold-Petry, Marcel F. Nold, Ofer Levy, Yossef Kliger, Anat Oren, Itamar Borukhov, Christoph Becker, Stefan Wirtz, Manjeet K. Sandhu, Markus Neurath, Charles A. Dinarello

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Background: The expression of heat shock protein gp96 is strongly correlated with the degree of tissue inflammation in ulcerative colitis and Crohn's disease, thereby leading us to the hypothesis that inhibition of expression via gp96-II peptide prevents intestinal inflammation. Methods: We employed daily injections of gp96-II peptide in two murine models of intestinal inflammation, the first resulting from five daily injections of IL-12/IL-18, the second via a single intrarectal application of TNBS (2,4,6-trinitrobenzenesulfonic acid). We also assessed the effectiveness of gp96-II peptide in murine and human primary cell culture. Results: In the IL-12/IL-18 model, all gp96-II peptide-treated animals survived until day 5, whereas 80% of placebo-injected animals died. gp96-II peptide reduced IL-12/IL-18-induced plasma IFγ by 89%, IL-1β by 63%, IL-6 by 43% and tumor necrosis factor (TNF) by 70% compared to controls. The clinical assessment Disease Activity Index of intestinal inflammation severity was found to be significantly lower in the gp96-II-treated animals when compared to vehicle-injected mice. gp96-II peptide treatment in the TNBS model limited weight loss to 5% on day 7 compared with prednisolone treatment, whereas placebo-treated animals suffered a 20% weight loss. Histological disease severity was reduced equally by prednisolone (by 40%) and gp96-II peptide (35%). Mice treated with either gp96-II peptide or prednisolone exhibited improved endoscopic scores compared with vehicle-treated control mice: vascularity, fibrin, granularity, and translucency scores were reduced by up to 49% by prednisolone and by up to 30% by gp96-II peptide. In vitro, gp96-II peptide reduced TLR2-, TLR4- and IL-12/IL-18-induced cytokine expression in murine splenocytes, with declines in constitutive IL-6 (54%), lipopolysaccharide-induced TNF (48%), IL-6 (81%) and in Staphylococcus epidermidis-induced TNF (67%) and IL-6 (81%), as well as IL-12/IL-18-induced IFNγ (75%). gp96-II peptide reduced IL-1β, IL-6, TNF and GM-CSF in human peripheral blood mononuclear cells to a similar degree without affecting cell viability, whereas RANTES, IL-25 and MIF were twofold to threefold increased. Conclusion: gp96-II peptide protects against murine intestinal inflammation by regulating inflammation in vivo and in vitro, pointing to its promise as a novel treatment for inflammatory bowel disease.

Original languageEnglish
Article number1531
Number of pages13
JournalFrontiers in Immunology
Volume8
Issue numberDEC
DOIs
Publication statusPublished - 11 Dec 2017

Keywords

  • Anti-inflammatory agent
  • Biologics
  • Cytokines and inflammation
  • Gp96
  • Immunemodulatory
  • Inflammatory bowel disease
  • Intestinal inflammation
  • Therapeutics

Cite this

Nold-Petry, Claudia A. ; Nold, Marcel F. ; Levy, Ofer ; Kliger, Yossef ; Oren, Anat ; Borukhov, Itamar ; Becker, Christoph ; Wirtz, Stefan ; Sandhu, Manjeet K. ; Neurath, Markus ; Dinarello, Charles A. / Gp96 peptide antagonist gp96-II confers therapeutic effects in murine intestinal inflammation. In: Frontiers in Immunology. 2017 ; Vol. 8, No. DEC.
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title = "Gp96 peptide antagonist gp96-II confers therapeutic effects in murine intestinal inflammation",
abstract = "Background: The expression of heat shock protein gp96 is strongly correlated with the degree of tissue inflammation in ulcerative colitis and Crohn's disease, thereby leading us to the hypothesis that inhibition of expression via gp96-II peptide prevents intestinal inflammation. Methods: We employed daily injections of gp96-II peptide in two murine models of intestinal inflammation, the first resulting from five daily injections of IL-12/IL-18, the second via a single intrarectal application of TNBS (2,4,6-trinitrobenzenesulfonic acid). We also assessed the effectiveness of gp96-II peptide in murine and human primary cell culture. Results: In the IL-12/IL-18 model, all gp96-II peptide-treated animals survived until day 5, whereas 80{\%} of placebo-injected animals died. gp96-II peptide reduced IL-12/IL-18-induced plasma IFγ by 89{\%}, IL-1β by 63{\%}, IL-6 by 43{\%} and tumor necrosis factor (TNF) by 70{\%} compared to controls. The clinical assessment Disease Activity Index of intestinal inflammation severity was found to be significantly lower in the gp96-II-treated animals when compared to vehicle-injected mice. gp96-II peptide treatment in the TNBS model limited weight loss to 5{\%} on day 7 compared with prednisolone treatment, whereas placebo-treated animals suffered a 20{\%} weight loss. Histological disease severity was reduced equally by prednisolone (by 40{\%}) and gp96-II peptide (35{\%}). Mice treated with either gp96-II peptide or prednisolone exhibited improved endoscopic scores compared with vehicle-treated control mice: vascularity, fibrin, granularity, and translucency scores were reduced by up to 49{\%} by prednisolone and by up to 30{\%} by gp96-II peptide. In vitro, gp96-II peptide reduced TLR2-, TLR4- and IL-12/IL-18-induced cytokine expression in murine splenocytes, with declines in constitutive IL-6 (54{\%}), lipopolysaccharide-induced TNF (48{\%}), IL-6 (81{\%}) and in Staphylococcus epidermidis-induced TNF (67{\%}) and IL-6 (81{\%}), as well as IL-12/IL-18-induced IFNγ (75{\%}). gp96-II peptide reduced IL-1β, IL-6, TNF and GM-CSF in human peripheral blood mononuclear cells to a similar degree without affecting cell viability, whereas RANTES, IL-25 and MIF were twofold to threefold increased. Conclusion: gp96-II peptide protects against murine intestinal inflammation by regulating inflammation in vivo and in vitro, pointing to its promise as a novel treatment for inflammatory bowel disease.",
keywords = "Anti-inflammatory agent, Biologics, Cytokines and inflammation, Gp96, Immunemodulatory, Inflammatory bowel disease, Intestinal inflammation, Therapeutics",
author = "Nold-Petry, {Claudia A.} and Nold, {Marcel F.} and Ofer Levy and Yossef Kliger and Anat Oren and Itamar Borukhov and Christoph Becker and Stefan Wirtz and Sandhu, {Manjeet K.} and Markus Neurath and Dinarello, {Charles A.}",
year = "2017",
month = "12",
day = "11",
doi = "10.3389/fimmu.2017.01531",
language = "English",
volume = "8",
journal = "Frontiers in Immunology",
issn = "1664-3224",
publisher = "Frontiers Media",
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}

Nold-Petry, CA, Nold, MF, Levy, O, Kliger, Y, Oren, A, Borukhov, I, Becker, C, Wirtz, S, Sandhu, MK, Neurath, M & Dinarello, CA 2017, 'Gp96 peptide antagonist gp96-II confers therapeutic effects in murine intestinal inflammation', Frontiers in Immunology, vol. 8, no. DEC, 1531. https://doi.org/10.3389/fimmu.2017.01531

Gp96 peptide antagonist gp96-II confers therapeutic effects in murine intestinal inflammation. / Nold-Petry, Claudia A.; Nold, Marcel F.; Levy, Ofer; Kliger, Yossef; Oren, Anat; Borukhov, Itamar; Becker, Christoph; Wirtz, Stefan; Sandhu, Manjeet K.; Neurath, Markus; Dinarello, Charles A.

In: Frontiers in Immunology, Vol. 8, No. DEC, 1531, 11.12.2017.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Nold-Petry, Claudia A.

AU - Nold, Marcel F.

AU - Levy, Ofer

AU - Kliger, Yossef

AU - Oren, Anat

AU - Borukhov, Itamar

AU - Becker, Christoph

AU - Wirtz, Stefan

AU - Sandhu, Manjeet K.

AU - Neurath, Markus

AU - Dinarello, Charles A.

PY - 2017/12/11

Y1 - 2017/12/11

N2 - Background: The expression of heat shock protein gp96 is strongly correlated with the degree of tissue inflammation in ulcerative colitis and Crohn's disease, thereby leading us to the hypothesis that inhibition of expression via gp96-II peptide prevents intestinal inflammation. Methods: We employed daily injections of gp96-II peptide in two murine models of intestinal inflammation, the first resulting from five daily injections of IL-12/IL-18, the second via a single intrarectal application of TNBS (2,4,6-trinitrobenzenesulfonic acid). We also assessed the effectiveness of gp96-II peptide in murine and human primary cell culture. Results: In the IL-12/IL-18 model, all gp96-II peptide-treated animals survived until day 5, whereas 80% of placebo-injected animals died. gp96-II peptide reduced IL-12/IL-18-induced plasma IFγ by 89%, IL-1β by 63%, IL-6 by 43% and tumor necrosis factor (TNF) by 70% compared to controls. The clinical assessment Disease Activity Index of intestinal inflammation severity was found to be significantly lower in the gp96-II-treated animals when compared to vehicle-injected mice. gp96-II peptide treatment in the TNBS model limited weight loss to 5% on day 7 compared with prednisolone treatment, whereas placebo-treated animals suffered a 20% weight loss. Histological disease severity was reduced equally by prednisolone (by 40%) and gp96-II peptide (35%). Mice treated with either gp96-II peptide or prednisolone exhibited improved endoscopic scores compared with vehicle-treated control mice: vascularity, fibrin, granularity, and translucency scores were reduced by up to 49% by prednisolone and by up to 30% by gp96-II peptide. In vitro, gp96-II peptide reduced TLR2-, TLR4- and IL-12/IL-18-induced cytokine expression in murine splenocytes, with declines in constitutive IL-6 (54%), lipopolysaccharide-induced TNF (48%), IL-6 (81%) and in Staphylococcus epidermidis-induced TNF (67%) and IL-6 (81%), as well as IL-12/IL-18-induced IFNγ (75%). gp96-II peptide reduced IL-1β, IL-6, TNF and GM-CSF in human peripheral blood mononuclear cells to a similar degree without affecting cell viability, whereas RANTES, IL-25 and MIF were twofold to threefold increased. Conclusion: gp96-II peptide protects against murine intestinal inflammation by regulating inflammation in vivo and in vitro, pointing to its promise as a novel treatment for inflammatory bowel disease.

AB - Background: The expression of heat shock protein gp96 is strongly correlated with the degree of tissue inflammation in ulcerative colitis and Crohn's disease, thereby leading us to the hypothesis that inhibition of expression via gp96-II peptide prevents intestinal inflammation. Methods: We employed daily injections of gp96-II peptide in two murine models of intestinal inflammation, the first resulting from five daily injections of IL-12/IL-18, the second via a single intrarectal application of TNBS (2,4,6-trinitrobenzenesulfonic acid). We also assessed the effectiveness of gp96-II peptide in murine and human primary cell culture. Results: In the IL-12/IL-18 model, all gp96-II peptide-treated animals survived until day 5, whereas 80% of placebo-injected animals died. gp96-II peptide reduced IL-12/IL-18-induced plasma IFγ by 89%, IL-1β by 63%, IL-6 by 43% and tumor necrosis factor (TNF) by 70% compared to controls. The clinical assessment Disease Activity Index of intestinal inflammation severity was found to be significantly lower in the gp96-II-treated animals when compared to vehicle-injected mice. gp96-II peptide treatment in the TNBS model limited weight loss to 5% on day 7 compared with prednisolone treatment, whereas placebo-treated animals suffered a 20% weight loss. Histological disease severity was reduced equally by prednisolone (by 40%) and gp96-II peptide (35%). Mice treated with either gp96-II peptide or prednisolone exhibited improved endoscopic scores compared with vehicle-treated control mice: vascularity, fibrin, granularity, and translucency scores were reduced by up to 49% by prednisolone and by up to 30% by gp96-II peptide. In vitro, gp96-II peptide reduced TLR2-, TLR4- and IL-12/IL-18-induced cytokine expression in murine splenocytes, with declines in constitutive IL-6 (54%), lipopolysaccharide-induced TNF (48%), IL-6 (81%) and in Staphylococcus epidermidis-induced TNF (67%) and IL-6 (81%), as well as IL-12/IL-18-induced IFNγ (75%). gp96-II peptide reduced IL-1β, IL-6, TNF and GM-CSF in human peripheral blood mononuclear cells to a similar degree without affecting cell viability, whereas RANTES, IL-25 and MIF were twofold to threefold increased. Conclusion: gp96-II peptide protects against murine intestinal inflammation by regulating inflammation in vivo and in vitro, pointing to its promise as a novel treatment for inflammatory bowel disease.

KW - Anti-inflammatory agent

KW - Biologics

KW - Cytokines and inflammation

KW - Gp96

KW - Immunemodulatory

KW - Inflammatory bowel disease

KW - Intestinal inflammation

KW - Therapeutics

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