Antagonism of the chemokine Ccl5 ameliorates experimental liver fibrosis in mice
Marie-Luise Berres, … , Christian Trautwein, Hermann E. Wasmuth
Marie-Luise Berres, … , Christian Trautwein, Hermann E. Wasmuth
Published October 18, 2010
Citation Information: J Clin Invest. 2010;120(11):4129-4140. https://doi.org/10.1172/JCI41732.
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Research Article Hepatology

Antagonism of the chemokine Ccl5 ameliorates experimental liver fibrosis in mice

Abstract

Activation of hepatic stellate cells in response to chronic inflammation represents a crucial step in the development of liver fibrosis. However, the molecules involved in the interaction between immune cells and stellate cells remain obscure. Herein, we identify the chemokine CCL5 (also known as RANTES), which is induced in murine and human liver after injury, as a central mediator of this interaction. First, we showed in patients with liver fibrosis that CCL5 haplotypes and intrahepatic CCL5 mRNA expression were associated with severe liver fibrosis. Consistent with this, we detected Ccl5 mRNA and CCL5 protein in 2 mouse models of liver fibrosis, induced by either injection of carbon tetrachloride (CCl4) or feeding on a methionine and choline–deficient (MCD) diet. In these models, Ccl5–/– mice exhibited decreased hepatic fibrosis, with reduced stellate cell activation and immune cell infiltration. Transplantation of Ccl5-deficient bone marrow into WT recipients attenuated liver fibrosis, identifying infiltrating hematopoietic cells as the main source of Ccl5. We then showed that treatment with the CCL5 receptor antagonist Met-CCL5 inhibited cultured stellate cell migration, proliferation, and chemokine and collagen secretion. Importantly, in vivo administration of Met-CCL5 greatly ameliorated liver fibrosis in mice and was able to accelerate fibrosis regression. Our results define a successful therapeutic approach to reduce experimental liver fibrosis by antagonizing Ccl5 receptors.

Authors

Marie-Luise Berres, Rory R. Koenen, Anna Rueland, Mirko Moreno Zaldivar, Daniel Heinrichs, Hacer Sahin, Petra Schmitz, Konrad L. Streetz, Thomas Berg, Nikolaus Gassler, Ralf Weiskirchen, Amanda Proudfoot, Christian Weber, Christian Trautwein, Hermann E. Wasmuth

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Figure 3

Experimental liver fibrosis in Ccl5-knockout mice.

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Experimental liver fibrosis in Ccl5-knockout mice. (A) Representativ...
(A) Representative Sirius red stainings of WT and Ccl5–/– mice after challenge with CCl4 (original magnification, ×40). Reduced fibrosis in Ccl5–/– mice (n = 12/group) is validated by the significantly lower Sirius red–positive area (***P < 0.001) and decreased hydroxyproline concentrations (*P < 0.05). (B) Ccl5–/– mice also have lower ALT values compared with WT littermates (**P < 0.01). (C) Treatment of Ccl5–/– mice with CCl4 leads to significantly reduced mRNA levels of Col1a1, Tgfb1, Timp1, and Il6 (all with P values of at least < 0.05; *P < 0.05, **P < 0.01, ***P < 0.001), compared with WT mice. (D) Immunohistochemistry demonstrates reduced α-SMA–positive cells within the livers of Ccl5–/– mice compared with WT mice after induction of liver fibrosis (original magnification, ×100). Decreased α-SMA protein expression is also evident in liver lysates of Ccl5–/– mice (representative samples). (E) Ameliorated fibrogenesis in Ccl5–/– mice (n = 12/group) is confirmed in the MCD diet model of liver fibrosis, as shown by representative Sirius red stainings (original magnification, ×40). Decreased deposition of collagen in Ccl5–/– animals is validated by the reduced Sirius red–positive area (***P < 0.001) and lower hydroxyproline concentrations (*P < 0.05). (F) As in the CCl4 model, ALT values are also reduced in Ccl5–/– mice after feeding with the MCD diet (**P < 0.01). (G) Likewise, Col1a1 and Timp1 mRNA is reduced in Ccl5–/– mice compared with their littermates after feedings with MCD diet for 8 weeks (*P < 0.05). (H) Furthermore, Ccl5–/– mice show a trend toward lower hepatic triglyceride levels (P = 0.1).