Identification of an atypical monocyte and committed progenitor involved in fibrosis (original) (raw)

References

  1. Asano, K. et al. CD169-positive macrophages dominate antitumor immunity by crosspresenting dead cell-associated antigens. Immunity 34, 85–95 (2011)
    Article CAS Google Scholar
  2. Auffray, C. et al. Monitoring of blood vessels and tissues by a population of monocytes with patrolling behavior. Science 317, 666–670 (2007)
    Article ADS CAS Google Scholar
  3. Kohyama, M. et al. Role for Spi-C in the development of red pulp macrophages and splenic iron homeostasis. Nature 457, 318–321 (2009)
    Article ADS CAS Google Scholar
  4. Okabe, Y. & Medzhitov, R. Tissue-specific signals control reversible program of localization and functional polarization of macrophages. Cell 157, 832–844 (2014)
    Article CAS Google Scholar
  5. Swirski, F. K. et al. Identification of splenic reservoir monocytes and their deployment to inflammatory sites. Science 325, 612–616 (2009)
    Article ADS CAS Google Scholar
  6. Duffield, J. S., Lupher, M., Thannickal, V. J. & Wynn, T. A. Host responses in tissue repair and fibrosis. Annu. Rev. Pathol. 8, 241–276 (2013)
    Article CAS Google Scholar
  7. Sica, A., Invernizzi, P. & Mantovani, A. Macrophage plasticity and polarization in liver homeostasis and pathology. Hepatology 59, 2034–2042 (2014)
    Article Google Scholar
  8. Wynn, T. A., Chawla, A. & Pollard, J. W. Macrophage biology in development, homeostasis and disease. Nature 496, 445–455 (2013)
    Article ADS CAS Google Scholar
  9. Sheel, M. & Engwerda, C. R. The diverse roles of monocytes in inflammation caused by protozoan parasitic diseases. Trends Parasitol. 28, 408–416 (2012)
    Article CAS Google Scholar
  10. Tsukui, T. et al. Qualitative rather than quantitative changes are hallmarks of fibroblasts in bleomycin-induced pulmonary fibrosis. Am. J. Pathol. 183, 758–773 (2013)
    Article CAS Google Scholar
  11. Yáñez, A., Ng, M. Y., Hassanzadeh-Kiabi, N. & Goodridge, H. S. IRF8 acts in lineage-committed rather than oligopotent progenitors to control neutrophil vs monocyte production. Blood 125, 1452–1459 (2015)
    Article Google Scholar
  12. Arinobu, Y., Iwasaki, H. & Akashi, K. Origin of basophils and mast cells. Allergol. Int. 58, 21–28 (2009)
    Article CAS Google Scholar
  13. Li, Y., Qi, X., Liu, B. & Huang, H. The STAT5–GATA2 pathway is critical in basophil and mast cell differentiation and maintenance. J. Immunol. 194, 4328–4338 (2015)
    Article CAS Google Scholar
  14. Nei, Y. et al. GATA-1 regulates the generation and function of basophils. Proc. Natl Acad. Sci. USA 110, 18620–18625 (2013)
    Article ADS CAS Google Scholar
  15. Scheller, M. et al. Altered development and cytokine responses of myeloid progenitors in the absence of transcription factor, interferon consensus sequence binding protein. Blood 94, 3764–3771 (1999)
    CAS Google Scholar
  16. Gutknecht, M. F. & Bouton, A. H. Functional significance of mononuclear phagocyte populations generated through adult hematopoiesis. J. Leukoc. Biol. 96, 969–980 (2014)
    Article Google Scholar
  17. Terry, R. L. & Miller, S. D. Molecular control of monocyte development. Cell. Immunol. 291, 16–21 (2014)
    Article CAS Google Scholar
  18. Yona, S. et al. Fate mapping reveals origins and dynamics of monocytes and tissue macrophages under homeostasis. Immunity 38, 79–91 (2013)
    Article CAS Google Scholar
  19. Auffray, C. et al. CX3CR1+CD115+CD135+ common macrophage/DC precursors and the role of CX3CR1 in their response to inflammation. J. Exp. Med. 206, 595–606 (2009)
    Article CAS Google Scholar
  20. Hettinger, J. et al. Origin of monocytes and macrophages in a committed progenitor. Nat. Immunol. 14, 821–830 (2013)
    Article CAS Google Scholar
  21. Satoh, T. et al. Critical role of Trib1 in differentiation of tissue-resident M2-like macrophages. Nature 495, 524–528 (2013)
    Article ADS CAS Google Scholar
  22. Satoh, T. et al. The Jmjd3-Irf4 axis regulates M2 macrophage polarization and host responses against helminth infection. Nat. Immunol. 11, 936–944 (2010)
    Article CAS Google Scholar
  23. Hanna, R. N. et al. The transcription factor NR4A1 (Nur77) controls bone marrow differentiation and the survival of Ly6C− monocytes. Nat. Immunol. 12, 778–785 (2011)
    Article CAS Google Scholar
  24. Tanaka, T. et al. Targeted disruption of the NF-IL6 gene discloses its essential role in bacteria killing and tumor cytotoxicity by macrophages. Cell 80, 353–361 (1995)
    Article CAS Google Scholar
  25. Tanaka, T., Yoshida, N., Kishimoto, T. & Akira, S. Defective adipocyte differentiation in mice lacking the C/EBPβ and/or C/EBPδ gene. EMBO J. 16, 7432–7443 (1997)
    Article CAS Google Scholar
  26. Omori, Y. et al. Mef2d is essential for the maturation and integrity of retinal photoreceptor and bipolar cells. Genes Cells 20, 408–426 (2015)
    Article CAS Google Scholar
  27. Aoyama, K., Takagi, T., Hirase, A. & Miyazawa, A. STEM tomography for thick biological specimens. Ultramicroscopy 109, 70–80 (2008)
    Article CAS Google Scholar
  28. Kremer, J. R., Mastronarde, D. N. & McIntosh, J. R. Computer visualization of three-dimensional image data using IMOD. J. Struct. Biol. 116, 71–76 (1996)
    Article CAS Google Scholar

Download references

Acknowledgements

We thank S. Saeki, S. Watanabe, R. Takenaka and M. Miyamoto for assistance with experiments, and T. Matsuki, T. Kawasaki, H. Kanemaru, H. Tanaka and K. Kuniyoshi for helpful discussions. We thank T. Kitamura for providing PlatE cells. We also thank E. Kamada for secretarial assistance, and C. Funamoto, N. Kitagaki, A. Wataki, K. Yokoyama and R. Kawaguchi for technical assistance. This work was supported by the Government of Japan and the Japan Society for the Promotion of Science (JSPS) through the funding program for World-Leading Innovative R&D on Science and Technology (FIRST Program), by Japan Science and Technology Agency (JST) thorough funding for a Grant-in-Aid for Young Scientists (A)(16H06234), Specially Promoted Research (15H05704) and the US National Institutes of Health (P01-AI070167) and ‘Visionary Research Fund’ from Takeda Science Foundation. A part of this work was supported by the Nanotechnology Platform (project number 12024046) of the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan.

Author information

Author notes

  1. Takashi Satoh and Katsuhiro Nakagawa: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Host Defense, World Premier Institute Immunology Frontier Research Center, Osaka University, Osaka, 565-0871, Japan
    Takashi Satoh, Katsuhiro Nakagawa, Fumihiro Yamane, Kiyoharu Fukushima, Isao Ebina & Shizuo Akira
  2. Department of Host Defense, Research Institute for Microbial Diseases (RIMD), Osaka University, Osaka, 565-0871, Japan
    Takashi Satoh, Katsuhiro Nakagawa, Fumihiro Yamane, Kiyoharu Fukushima, Isao Ebina & Shizuo Akira
  3. Laboratory of Biofunctional Imaging, World Premier Institute Immunology Frontier Research Center, Osaka University, Osaka, 565-0871, Japan
    Fuminori Sugihara & Yoshichika Yoshioka
  4. Research Center for Ultra-high Voltage Electron Microscopy, Osaka University, Osaka, 567-0047, Japan
    Ryusuke Kuwahara
  5. Discovery Research Department, Research Division, Chugai Pharmaceutical Co., Ltd., 247-8530, Kanagawa, Japan
    Motooki Ashihara, Yosuke Minowa & Isao Ebina
  6. Core Research for Evolutional Science and Technology, Japan Agency for Medical Research and Development, Tokyo, 100-0004, Japan
    Atsushi Kumanogoh
  7. Department of Respiratory Medicine, Allergy and Rheumatic Diseases, Graduate School of Medicine, Osaka University, Osaka, 565-0871, Japan
    Atsushi Kumanogoh

Authors

  1. Takashi Satoh
  2. Katsuhiro Nakagawa
  3. Fuminori Sugihara
  4. Ryusuke Kuwahara
  5. Motooki Ashihara
  6. Fumihiro Yamane
  7. Yosuke Minowa
  8. Kiyoharu Fukushima
  9. Isao Ebina
  10. Yoshichika Yoshioka
  11. Atsushi Kumanogoh
  12. Shizuo Akira

Contributions

T.S. designed and performed the experiments and wrote the manuscript. K.F., I.E., F.Y. and A.K. helped with experiments. K.N. performed the experiments. M.A. and Y.M. performed bioinformatics analysis. F.S. and Y.Y. performed the MRI analysis. R.K. performed the electron microscopy analysis. S.A. wrote the manuscript and supervised the project.

Corresponding author

Correspondence toShizuo Akira.

Ethics declarations

Competing interests

S.A. has research support from Chugai Pharmaceutical Co., Ltd. The terms of this arrangement have been reviewed and approved by the Osaka University in accordance with its policy on objectivity in research.

Additional information

Reviewer Information Nature thanks I. Amit, D. Brenner, F. Geissmann and the other anonymous reviewer(s) for their contribution to the peer review of this work.

Extended data figures and tables

Extended Data Figure 1 Characterization of various macrophage/monocytes.

a, Indicated cell types were sorted and RNA was collected from them. Each cell type was subjected to microarray analysis. Principal component analysis by using gene expression data was performed. b, c, Indicated cell types in bone marrow were sorted, and each cell type was adoptively transferred into the wild-type mice, which were administered bleomycin. Images of Azan staining (b) and quantity of hydroxyproline are shown (c). Similar results were obtained in three independent experiments (a–c). Scale bars, 100 μm. *P < 0.05, **P < 0.01.

Extended Data Figure 2 _Cebpb_−/− chimaeras are resistant to the development of fibrosis.

a, Quantity of hydroxyproline 14 days after bleomycin administration was determined. b, Indicated cytokines in the BAL 3 days after bleomycin administration were determined by ELISA. c, d, Images of H&E staining in the lung tissue 4 days (c) and 13 days (d) after intratracheal injection with bleomycin. Scale bars, 50 μm. e, MRI imaging, H&E, Azan and Sirius red staining of lung. f, MRI images of T2 weighted (T2W), T1 weighted (T1W) and water suppression (WS) was shown (left panels). Azan staining of fibrotic lungs and corresponding schematic images of fibrotic region (right panels). g, Indicated cell types in bone marrow were sorted, and adoptively transferred into the wild-type mice with bleomycin administration. Quantity of hydroxyproline was shown. Similar results were obtained in three independent experiments (a–g). *P < 0.05, **P < 0.01.

Extended Data Figure 3 Cebpd is dispensable for fibrosis and SatM differentiation.

a, The survival rate of chimaeras (at least n = 5) inoculated with bleomycin (3 μg per g body weight). Similar results were obtained in three independent experiments. b, The proportions of SatM in bone marrow (upper left panel), spleen (bottom left panel), blood (upper right panel) and lung (bottom right panel) were shown. Similar results were obtained in five independent experiments.

Extended Data Figure 4 Modified high-fat diet liver fibrosis was repressed in _Cebpb_−/− chimaeras.

a, Liver was collected from wild-type and _Cebpb_−/− chimaeric mice fed on CDA-HFD, and indicated mRNA were determined by qPCR. b, Images of Azan staining of liver tissue. Scale bar, 50 μm. c, Quantity of hydroxyproline was determined. Similar results were obtained in three independent experiments (a–g). *P < 0.05, **P < 0.01.

Extended Data Figure 5 Wound healing progressed normally in _Cebpb_−/− chimaeras.

a, b, Chimaeric mice with wild-type and _Cebpb_−/− cells (at least n = 5) were injured by biopsy (diameter (Φ) = 6 mm). The repair rate of the wound area was monitored. c, H&E analysis of skin was performed. Similar results were obtained in three independent experiments.

Extended Data Figure 6 FACS analysis of various immune cells in _Cebpb_−/− chimaeras.

a, FACS analysis of spleen. The proportions of neutrophils (upper left), eosinophils (middle left), natural killer cells (bottom left), B cells, T cells (upper right), plasmacytoid dendritic cells (pDC) and conventional dendritic cells (cDC) were shown. b, FACS analysis of SatM. The proportions of these cells in blood (left panel), spleen (centre panel) and lung (right panel) were shown. Similar results were obtained in five independent experiments (a, b).

Extended Data Figure 7 Initiation factor of fibrosis produced from activated SatM.

a, Sorted SatM were cultured with Toll-like receptor ligands for 48 h, and concentrations of indicated cytokines were determined by ELISA. b, Fibroblasts were cultured with TNF-α for indicated time, and expression level of Spp1 mRNA was determined by qPCR. Similar results were obtained in three independent experiments (a, b). *P < 0.05, **P < 0.01.

Extended Data Figure 8 Characterization of SatM.

a, The FSC–SSC profiles of whole-cell and SatM in bone marrow (left and centre panels) and merged image (right panel) were shown. Similar results were obtained in three independent experiments. b, Expression of phosphorylated (p-) and unphosphorylated Erk and Akt and actin in SatM stimulated with M-CSF (50 ng ml−1). c, Flow cytometric analysis of spleen. The proportions of SatM were shown by F4/80−, Mac1+, Ly6C−, Msr1+ and Ceacam1+. Similar results were obtained in five independent experiments (b, c). d, The expression level of indicated cell surface molecules of SatM from bone marrow were shown. Similar results were obtained in three independent experiments. e, Cell lysates were prepared from SatM in wild-type mice, and were subjected to whole-cell proteomics analysis. Substantially expressed granule proteins were described. f, g, Sorted Siglec-F+CCR3+CD4− population, Ly6G+Mac1+ population and DX5+FcεRI+c-kit−CD3−CD19− population were stained with Diff-Quick after cytospin centrifugation and were photographed (f) and investigated by TEM (g). Scale bars were shown in indicated images. Similar results were obtained in three independent experiments (f, g). h, Flow cytometric analysis of spleen obtained from wild-type and ΔdblGATA mutant mice. The proportions of eosinophils (upper panel) and SatM (bottom panel) were shown. Similar results were obtained in five independent experiments.

Extended Data Figure 9 Characterization of various progenitors.

a, The proportion of GMPs (left panel) and MDPs (right panel) were shown. Similar results were obtained in three independent experiments. b, Sorted Ly6C−FcεRI+ GMP population were stained with Diff-Quick after cytospin centrifugation and were photographed. Similar results were obtained in five independent experiments. c, Principal component analysis by using gene expression of indicated cells was performed. d, SatM, inflammatory monocytes, SMPs, cMoPs, MDPs and GMPs were sorted and subjected to microarray analysis. Data on transcription factor genes were partitioned by _k_-means clustering using the threshold k = 8. Clustered molecules are shown. e, Sorted GFP+Lin–c-kit–C5aR+CD115+FcεRI+Ly6C– populations were transferred into wild-type mice, and then analysed by FACS. Similar results were obtained in three independent experiments.

Extended Data Figure 10 Macrophages regulated by Jmjd3 and Trib1 were dispensable for the development of fibrosis.

a, The proportion of SatM in _Jmjd3_−/− (left panel) and _Trib1_−/− (right panel) were shown. Similar results were obtained in five independent experiments. b, c, Chimaeric mice with wild-type, _Jmjd3_−/− or _Trib1_−/− haematopoietic cells (at least n = 5) were intratracheally inoculated with bleomycin (3 μg per g body weight). The survival rate of mice was monitored (b). Images of Azan staining for collagen fibres in the lung tissue of wild-type, _Jmjd3_−/− and _Trib1_−/− bone marrow chimaeric mice (c). Similar results were obtained in three independent experiments.

Supplementary information

PowerPoint slides

Rights and permissions

About this article

Cite this article

Satoh, T., Nakagawa, K., Sugihara, F. et al. Identification of an atypical monocyte and committed progenitor involved in fibrosis.Nature 541, 96–101 (2017). https://doi.org/10.1038/nature20611

Download citation