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 6

In vivo inhibition of liver fibrosis by Met-CCL5.

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In vivo inhibition of liver fibrosis by Met-CCL5. (A) Representative Sir...
(A) Representative Sirius red stainings of CCl4-treated C57BL/6 WT mice, with a concomitant administration of vehicle or Met-CCL5 (n = 10/group; original magnification, ×40). (B) Reduced fibrosis in Met-CCL5–treated mice is validated by a significantly lower Sirius red–positive area (*P < 0.05) and decreased concentrations of hydroxyproline (**P < 0.01). (C) The reduced fibrotic response in Met-CCL5–treated mice is further evidenced by significantly reduced Col1a1 and Timp1 mRNA levels (*P < 0.05, **P < 0.01). (D) Representative FACS blot of NK1.1 and CD3-positive cell infiltration in vehicle- and Met-CCL5–treated animals, demonstrating a reduced T cell infiltration into the livers of Met-CCL5–treated mice, while numbers of NK and NKT cells are not significantly altered. (E) Statistical analysis of T cell infiltration (FACS) reveals a significantly reduced number of CD3- and, specifically, CD8-positive cells after treatment with Met-CCL5 (***P < 0.001). (F) In addition, numbers of CD68+-positive macrophages are also reduced in Met-CCL5–treated animals compared with those of vehicle-treated mice (*P < 0.05, n = 10/group). (G) The antifibrotic potential of Met-CCL5 is validated in the MCD fibrosis model by histology (original magnification, ×40). (H) Reduced liver damage in Met-CCL5–treated mice is evidenced by a lower Sirius red–positive area in histology (**P < 0.01) and significantly reduced hydroxyproline levels (**P < 0.01) compared with those of vehicle-treated animals.