Limits...
Human amniotic epithelial cell transplantation induces markers of alternative macrophage activation and reduces established hepatic fibrosis.

Manuelpillai U, Lourensz D, Vaghjiani V, Tchongue J, Lacey D, Tee JY, Murthi P, Chan J, Hodge A, Sievert W - PLoS ONE (2012)

Bottom Line: Transplantation of human amniotic epithelial cells (hAEC) from term delivered placenta has been shown to decrease mild to moderate hepatic fibrosis in a murine model.CCl(4) administration caused hepatic T cell infiltration that decreased significantly following hAEC transplantation.Alternatively activated M2 macrophages are associated with fibrosis resolution.

View Article: PubMed Central - PubMed

Affiliation: Center for Reproduction and Development, Monash Institute of Medical Research, Monash University, Melbourne, Australia.

ABSTRACT
Chronic hepatic inflammation from multiple etiologies leads to a fibrogenic response that can progress to cirrhosis and liver failure. Transplantation of human amniotic epithelial cells (hAEC) from term delivered placenta has been shown to decrease mild to moderate hepatic fibrosis in a murine model. To model advanced human liver disease and assess the efficacy of hAEC therapy, we transplanted hAEC in mice with advanced hepatic fibrosis. Immunocompetent C57BL/6 mice were administered carbon tetrachloride (CCl(4)) twice weekly resulting in bridging fibrosis by 12 weeks. hAEC (2 × 10(6)) were infused via the tail vein at week 8 or weeks 8 and 10 (single and double dose, respectively). Human cells were detected in mouse liver four weeks after transplantation showing hAEC engraftment. CCl(4) treated mice receiving single or double hAEC doses showed a significant but similar decrease in liver fibrosis area associated with decreased activation of collagen-producing hepatic stellate cells and decreased hepatic protein levels of the pro-fibrogenic cytokine, transforming growth factor-beta1. CCl(4) administration caused hepatic T cell infiltration that decreased significantly following hAEC transplantation. Hepatic macrophages play a crucial role in both fibrogenesis and fibrosis resolution. Mice exposed to CCl(4) demonstrated increased numbers of hepatic macrophages compared to normal mice; the number of macrophages decreased significantly in CCl(4) treated mice given hAEC. These mice had significantly lower hepatic protein levels of the chemokine monocyte chemoattractant protein-1 than mice given CCl(4) alone. Alternatively activated M2 macrophages are associated with fibrosis resolution. CCl(4) treated mice given hAEC showed increased expression of genes associated with M2 macrophages including YM-1, IL-10 and CD206. We provide novel data showing that hAEC transplantation induces a wound healing M2 macrophage phenotype associated with reduction of established hepatic fibrosis that justifies further investigation of this potential cell-based therapy for advanced hepatic fibrosis.

Show MeSH

Related in: MedlinePlus

Human amniotic epithelial cell transplantation induces expression of genes associated with alternately activated (M2) macrophages.mRNA expression of classically (M1) and alternately activated M2 macrophage genes were analyzed by real time quantitative PCR and the fold change calculated by the ΔΔCT method. Expression of M2 associated YM-1, CD206 and IL-10 was significantly elevated in livers of mice treated with hAEC compared to animals given carbon tetrachloride (CCl4) alone (A). M1 associated genes CCL17 and CCL5 were expressed but levels did not alter with hAEC treatment (B). IL-12b:IL-10 ratio was skewed towards an M2 phenotype. MMP-9 increased and MMP-12 expression decreased with hAEC treatment (C). *, ** and *** P<0.05, 0.01 and 0.001, respectively.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3375296&req=5

pone-0038631-g004: Human amniotic epithelial cell transplantation induces expression of genes associated with alternately activated (M2) macrophages.mRNA expression of classically (M1) and alternately activated M2 macrophage genes were analyzed by real time quantitative PCR and the fold change calculated by the ΔΔCT method. Expression of M2 associated YM-1, CD206 and IL-10 was significantly elevated in livers of mice treated with hAEC compared to animals given carbon tetrachloride (CCl4) alone (A). M1 associated genes CCL17 and CCL5 were expressed but levels did not alter with hAEC treatment (B). IL-12b:IL-10 ratio was skewed towards an M2 phenotype. MMP-9 increased and MMP-12 expression decreased with hAEC treatment (C). *, ** and *** P<0.05, 0.01 and 0.001, respectively.

Mentions: The polarization of macrophages to a M2 alternatively activated phenotype is associated with hepatic wound healing [30]. We therefore examined the phenotype of the hepatic macrophage population by determining gene expression of known macrophage associated markers in whole liver extracts. We found that CCl4-treated mice which had been infused with hAEC had increased mRNA expression of M2 associated markers YM-1 (P<0.05), CD206 and IL-10 (P<0.01; Figure 4A) compared with mice given CCl4 alone. Cnrip-1 expression also increased in hAEC treated mice but did not reach significance while Arg-1 and Cadm-1 mRNA expression remained unaltered. We also examined the expression of classically activated M1 macrophages that have been linked to fibrosis progression [30]. We found no change in the expression of M1 associated markers CCL17 and CCL5 in hAEC treated mice compared with mice given CCl4 alone (Figure 4B). We also examined the ratio of IL-12b mRNA, which is expressed by M1 macrophages [31], to IL-10 mRNA to determine whether the macrophage phenotype was skewed towards M1 or M2. hAEC treated mice had increased IL-10 mRNA expression and a significant decrease in the IL-12b:IL-10 ratio consistent with skewing towards M2 macrophages (P<0.001 and <0.05 for single and double doses respectively; Figure 4C). Macrophages express MMP-9 and MMP-12, which are important regulators of fibrolysis [27], [31]. MMP-9 expression was elevated while MMP-12 decreased in hAEC treated mice (P<0.05; Figure 4C).


Human amniotic epithelial cell transplantation induces markers of alternative macrophage activation and reduces established hepatic fibrosis.

Manuelpillai U, Lourensz D, Vaghjiani V, Tchongue J, Lacey D, Tee JY, Murthi P, Chan J, Hodge A, Sievert W - PLoS ONE (2012)

Human amniotic epithelial cell transplantation induces expression of genes associated with alternately activated (M2) macrophages.mRNA expression of classically (M1) and alternately activated M2 macrophage genes were analyzed by real time quantitative PCR and the fold change calculated by the ΔΔCT method. Expression of M2 associated YM-1, CD206 and IL-10 was significantly elevated in livers of mice treated with hAEC compared to animals given carbon tetrachloride (CCl4) alone (A). M1 associated genes CCL17 and CCL5 were expressed but levels did not alter with hAEC treatment (B). IL-12b:IL-10 ratio was skewed towards an M2 phenotype. MMP-9 increased and MMP-12 expression decreased with hAEC treatment (C). *, ** and *** P<0.05, 0.01 and 0.001, respectively.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3375296&req=5

pone-0038631-g004: Human amniotic epithelial cell transplantation induces expression of genes associated with alternately activated (M2) macrophages.mRNA expression of classically (M1) and alternately activated M2 macrophage genes were analyzed by real time quantitative PCR and the fold change calculated by the ΔΔCT method. Expression of M2 associated YM-1, CD206 and IL-10 was significantly elevated in livers of mice treated with hAEC compared to animals given carbon tetrachloride (CCl4) alone (A). M1 associated genes CCL17 and CCL5 were expressed but levels did not alter with hAEC treatment (B). IL-12b:IL-10 ratio was skewed towards an M2 phenotype. MMP-9 increased and MMP-12 expression decreased with hAEC treatment (C). *, ** and *** P<0.05, 0.01 and 0.001, respectively.
Mentions: The polarization of macrophages to a M2 alternatively activated phenotype is associated with hepatic wound healing [30]. We therefore examined the phenotype of the hepatic macrophage population by determining gene expression of known macrophage associated markers in whole liver extracts. We found that CCl4-treated mice which had been infused with hAEC had increased mRNA expression of M2 associated markers YM-1 (P<0.05), CD206 and IL-10 (P<0.01; Figure 4A) compared with mice given CCl4 alone. Cnrip-1 expression also increased in hAEC treated mice but did not reach significance while Arg-1 and Cadm-1 mRNA expression remained unaltered. We also examined the expression of classically activated M1 macrophages that have been linked to fibrosis progression [30]. We found no change in the expression of M1 associated markers CCL17 and CCL5 in hAEC treated mice compared with mice given CCl4 alone (Figure 4B). We also examined the ratio of IL-12b mRNA, which is expressed by M1 macrophages [31], to IL-10 mRNA to determine whether the macrophage phenotype was skewed towards M1 or M2. hAEC treated mice had increased IL-10 mRNA expression and a significant decrease in the IL-12b:IL-10 ratio consistent with skewing towards M2 macrophages (P<0.001 and <0.05 for single and double doses respectively; Figure 4C). Macrophages express MMP-9 and MMP-12, which are important regulators of fibrolysis [27], [31]. MMP-9 expression was elevated while MMP-12 decreased in hAEC treated mice (P<0.05; Figure 4C).

Bottom Line: Transplantation of human amniotic epithelial cells (hAEC) from term delivered placenta has been shown to decrease mild to moderate hepatic fibrosis in a murine model.CCl(4) administration caused hepatic T cell infiltration that decreased significantly following hAEC transplantation.Alternatively activated M2 macrophages are associated with fibrosis resolution.

View Article: PubMed Central - PubMed

Affiliation: Center for Reproduction and Development, Monash Institute of Medical Research, Monash University, Melbourne, Australia.

ABSTRACT
Chronic hepatic inflammation from multiple etiologies leads to a fibrogenic response that can progress to cirrhosis and liver failure. Transplantation of human amniotic epithelial cells (hAEC) from term delivered placenta has been shown to decrease mild to moderate hepatic fibrosis in a murine model. To model advanced human liver disease and assess the efficacy of hAEC therapy, we transplanted hAEC in mice with advanced hepatic fibrosis. Immunocompetent C57BL/6 mice were administered carbon tetrachloride (CCl(4)) twice weekly resulting in bridging fibrosis by 12 weeks. hAEC (2 × 10(6)) were infused via the tail vein at week 8 or weeks 8 and 10 (single and double dose, respectively). Human cells were detected in mouse liver four weeks after transplantation showing hAEC engraftment. CCl(4) treated mice receiving single or double hAEC doses showed a significant but similar decrease in liver fibrosis area associated with decreased activation of collagen-producing hepatic stellate cells and decreased hepatic protein levels of the pro-fibrogenic cytokine, transforming growth factor-beta1. CCl(4) administration caused hepatic T cell infiltration that decreased significantly following hAEC transplantation. Hepatic macrophages play a crucial role in both fibrogenesis and fibrosis resolution. Mice exposed to CCl(4) demonstrated increased numbers of hepatic macrophages compared to normal mice; the number of macrophages decreased significantly in CCl(4) treated mice given hAEC. These mice had significantly lower hepatic protein levels of the chemokine monocyte chemoattractant protein-1 than mice given CCl(4) alone. Alternatively activated M2 macrophages are associated with fibrosis resolution. CCl(4) treated mice given hAEC showed increased expression of genes associated with M2 macrophages including YM-1, IL-10 and CD206. We provide novel data showing that hAEC transplantation induces a wound healing M2 macrophage phenotype associated with reduction of established hepatic fibrosis that justifies further investigation of this potential cell-based therapy for advanced hepatic fibrosis.

Show MeSH
Related in: MedlinePlus