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Natural Killer Cells-Produced IFN-γ Improves Bone Marrow-Derived Hepatocytes Regeneration in Murine Liver Failure Model.

Li L, Zeng Z, Qi Z, Wang X, Gao X, Wei H, Sun R, Tian Z - Sci Rep (2015)

Bottom Line: Hepatic NK cells became activated during BMDH generation and were the major IFN-γ producers.Indeed, both NK cells and IFN-γ were required for BMDH generation since WT, but not NK-, IFN-γ-, or IFN-γR1-deficient BM transplantation successfully generated BMDHs and rescued survival in Fah(-/-) hosts.BM-derived myelomonocytes were determined to be the IFN-γ-responding cells.

View Article: PubMed Central - PubMed

Affiliation: Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230027, China.

ABSTRACT
Bone-marrow transplantation (BMT) can repopulate the liver through BM-derived hepatocyte (BMDH) generation, although the underlying mechanism remains unclear. Using fumarylacetoacetate hydrolase-deficient (Fah(-/-)) mice as a liver-failure model, we confirmed that BMDHs were generated by fusion of BM-derived CD11b(+)F4/80(+)myelomonocytes with resident Fah(-/-) hepatocytes. Hepatic NK cells became activated during BMDH generation and were the major IFN-γ producers. Indeed, both NK cells and IFN-γ were required for BMDH generation since WT, but not NK-, IFN-γ-, or IFN-γR1-deficient BM transplantation successfully generated BMDHs and rescued survival in Fah(-/-) hosts. BM-derived myelomonocytes were determined to be the IFN-γ-responding cells. The IFN-γ-IFN-γR interaction contributed to the myelomonocyte-hepatocyte fusion process, as most of the CD11b(+) BMDHs in mixed BM chimeric Fah(-/-) hosts transplanted with a 1:1 ratio of CD45.1(+) WT and CD45.2(+) Ifngr1(-/-) BM cells were of CD45.1(+) WT origin. Confirming these findings in vitro, IFN-γ dose-dependently promoted the fusion of GFP(+) myelomonocytes with Fah(-/-) hepatocytes due to a direct effect on myelomonocytes; similar results were observed using activated NK cells. In conclusion, BMDH generation requires NK cells to facilitate myelomonocyte-hepatocyte fusion in an IFN-γ-dependent manner, providing new insights for treating severe liver failure.

No MeSH data available.


Related in: MedlinePlus

BMT rescues liver failure through the generation of hepatocytes in Fah−/− mice.Survival rate (a) and body weight (b) of Fah−/− mice transplanted with Fah−/− (n = 4) or WT (n = 5) BMCs. (c) Total bilirubin levels in sera of Fah−/− mice transplanted with WT BMCs after NTBC withdrawal. (d,e) Liver tissues of WT BM-transplanted Fah−/− mice were collected 20 weeks after NTBC withdrawal and stained for Fah by immunohistochemistry (d, scale bar, 200 μm) and immunofluorescence (e, scale bar, 50 μm). The boundary of Fah+ hepatocyte area is indicated by dashed white line. (f) FACS analysis of BMDHs and Fah−/− hepatocytes 27 weeks after NTBC withdrawal. Gray shadows represent staining by the isotype control. Data are expressed as the mean ± SEM.
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f1: BMT rescues liver failure through the generation of hepatocytes in Fah−/− mice.Survival rate (a) and body weight (b) of Fah−/− mice transplanted with Fah−/− (n = 4) or WT (n = 5) BMCs. (c) Total bilirubin levels in sera of Fah−/− mice transplanted with WT BMCs after NTBC withdrawal. (d,e) Liver tissues of WT BM-transplanted Fah−/− mice were collected 20 weeks after NTBC withdrawal and stained for Fah by immunohistochemistry (d, scale bar, 200 μm) and immunofluorescence (e, scale bar, 50 μm). The boundary of Fah+ hepatocyte area is indicated by dashed white line. (f) FACS analysis of BMDHs and Fah−/− hepatocytes 27 weeks after NTBC withdrawal. Gray shadows represent staining by the isotype control. Data are expressed as the mean ± SEM.

Mentions: In order to begin exploring the mechanism underlying how BMDHs develop and function to repopulate the liver after BMT to treat hepatic failure, Fah−/− mice were transplanted with WT BMCs and monitored for more than 30 weeks after NTBC withdrawal. WT, but not Fah−/−, BMCs promoted survival (Fig. 1a), rescued severe weight loss (Fig. 1b), and decreased the elevated levels of total bilirubin (Fig. 1c) in Fah−/− mice, indicating that hepatocytes were directly and efficiently re-established in these mice after transplantation of WT BMCs. Indeed, robust formation of BM-derived, Fah+ hepatocytes was consistently observed in the livers of transplanted Fah−/− mice after NTBC withdrawal (Fig. 1d). These BMDHs were likely generated by fusion of donor WT BM-derived myelomonocytes with resident Fah−/− hepatocytes, as they expressed both myelomonocyte markers (CD45, CD11b) and Fah (Fig. 1e and Supplementary Fig. S1). In addition, BMDHs exhibited a CD11b+Ly6C–Ly6G–F4/80+CD11c– phenotype (Fig. 1f), identifying that liver-resident macrophages might be the major component within the myelomonocyte compartment to fuse with hepatocytes and allow their liver repopulation and survival.


Natural Killer Cells-Produced IFN-γ Improves Bone Marrow-Derived Hepatocytes Regeneration in Murine Liver Failure Model.

Li L, Zeng Z, Qi Z, Wang X, Gao X, Wei H, Sun R, Tian Z - Sci Rep (2015)

BMT rescues liver failure through the generation of hepatocytes in Fah−/− mice.Survival rate (a) and body weight (b) of Fah−/− mice transplanted with Fah−/− (n = 4) or WT (n = 5) BMCs. (c) Total bilirubin levels in sera of Fah−/− mice transplanted with WT BMCs after NTBC withdrawal. (d,e) Liver tissues of WT BM-transplanted Fah−/− mice were collected 20 weeks after NTBC withdrawal and stained for Fah by immunohistochemistry (d, scale bar, 200 μm) and immunofluorescence (e, scale bar, 50 μm). The boundary of Fah+ hepatocyte area is indicated by dashed white line. (f) FACS analysis of BMDHs and Fah−/− hepatocytes 27 weeks after NTBC withdrawal. Gray shadows represent staining by the isotype control. Data are expressed as the mean ± SEM.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4561890&req=5

f1: BMT rescues liver failure through the generation of hepatocytes in Fah−/− mice.Survival rate (a) and body weight (b) of Fah−/− mice transplanted with Fah−/− (n = 4) or WT (n = 5) BMCs. (c) Total bilirubin levels in sera of Fah−/− mice transplanted with WT BMCs after NTBC withdrawal. (d,e) Liver tissues of WT BM-transplanted Fah−/− mice were collected 20 weeks after NTBC withdrawal and stained for Fah by immunohistochemistry (d, scale bar, 200 μm) and immunofluorescence (e, scale bar, 50 μm). The boundary of Fah+ hepatocyte area is indicated by dashed white line. (f) FACS analysis of BMDHs and Fah−/− hepatocytes 27 weeks after NTBC withdrawal. Gray shadows represent staining by the isotype control. Data are expressed as the mean ± SEM.
Mentions: In order to begin exploring the mechanism underlying how BMDHs develop and function to repopulate the liver after BMT to treat hepatic failure, Fah−/− mice were transplanted with WT BMCs and monitored for more than 30 weeks after NTBC withdrawal. WT, but not Fah−/−, BMCs promoted survival (Fig. 1a), rescued severe weight loss (Fig. 1b), and decreased the elevated levels of total bilirubin (Fig. 1c) in Fah−/− mice, indicating that hepatocytes were directly and efficiently re-established in these mice after transplantation of WT BMCs. Indeed, robust formation of BM-derived, Fah+ hepatocytes was consistently observed in the livers of transplanted Fah−/− mice after NTBC withdrawal (Fig. 1d). These BMDHs were likely generated by fusion of donor WT BM-derived myelomonocytes with resident Fah−/− hepatocytes, as they expressed both myelomonocyte markers (CD45, CD11b) and Fah (Fig. 1e and Supplementary Fig. S1). In addition, BMDHs exhibited a CD11b+Ly6C–Ly6G–F4/80+CD11c– phenotype (Fig. 1f), identifying that liver-resident macrophages might be the major component within the myelomonocyte compartment to fuse with hepatocytes and allow their liver repopulation and survival.

Bottom Line: Hepatic NK cells became activated during BMDH generation and were the major IFN-γ producers.Indeed, both NK cells and IFN-γ were required for BMDH generation since WT, but not NK-, IFN-γ-, or IFN-γR1-deficient BM transplantation successfully generated BMDHs and rescued survival in Fah(-/-) hosts.BM-derived myelomonocytes were determined to be the IFN-γ-responding cells.

View Article: PubMed Central - PubMed

Affiliation: Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230027, China.

ABSTRACT
Bone-marrow transplantation (BMT) can repopulate the liver through BM-derived hepatocyte (BMDH) generation, although the underlying mechanism remains unclear. Using fumarylacetoacetate hydrolase-deficient (Fah(-/-)) mice as a liver-failure model, we confirmed that BMDHs were generated by fusion of BM-derived CD11b(+)F4/80(+)myelomonocytes with resident Fah(-/-) hepatocytes. Hepatic NK cells became activated during BMDH generation and were the major IFN-γ producers. Indeed, both NK cells and IFN-γ were required for BMDH generation since WT, but not NK-, IFN-γ-, or IFN-γR1-deficient BM transplantation successfully generated BMDHs and rescued survival in Fah(-/-) hosts. BM-derived myelomonocytes were determined to be the IFN-γ-responding cells. The IFN-γ-IFN-γR interaction contributed to the myelomonocyte-hepatocyte fusion process, as most of the CD11b(+) BMDHs in mixed BM chimeric Fah(-/-) hosts transplanted with a 1:1 ratio of CD45.1(+) WT and CD45.2(+) Ifngr1(-/-) BM cells were of CD45.1(+) WT origin. Confirming these findings in vitro, IFN-γ dose-dependently promoted the fusion of GFP(+) myelomonocytes with Fah(-/-) hepatocytes due to a direct effect on myelomonocytes; similar results were observed using activated NK cells. In conclusion, BMDH generation requires NK cells to facilitate myelomonocyte-hepatocyte fusion in an IFN-γ-dependent manner, providing new insights for treating severe liver failure.

No MeSH data available.


Related in: MedlinePlus