Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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GM-CSF Grown Bone Marrow Derived Cells Are Composed of Phenotypically Different Dendritic Cells and Macrophages

  • Na, Yi Rang (Department of Microbiology and Immunology, and Institute of Endemic Disease, Seoul National University College of Medicine) ;
  • Jung, Daun (Department of Microbiology and Immunology, and Institute of Endemic Disease, Seoul National University College of Medicine) ;
  • Gu, Gyo Jeong (Department of Microbiology and Immunology, and Institute of Endemic Disease, Seoul National University College of Medicine) ;
  • Seok, Seung Hyeok (Department of Microbiology and Immunology, and Institute of Endemic Disease, Seoul National University College of Medicine)
  • Received : 2016.06.28
  • Accepted : 2016.09.19
  • Published : 2016.10.31
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Abstract

Granulocyte-macrophage colony stimulating factor (GM-CSF) has a role in inducing emergency hematopoiesis upon exposure to inflammatory stimuli. Although GM-CSF generated murine bone marrow derived cells have been widely used as macrophages or dendritic cells in research, the exact characteristics of each cell population have not yet been defined. Here we discriminated GM-CSF grown bone marrow derived macrophages (GM-BMMs) from dendritic cells (GM-BMDCs) in several criteria. After C57BL/6J mice bone marrow cell culture for 7 days with GM-CSF supplementation, two main populations were observed in the attached cells based on MHCII and F4/80 marker expressions. GM-BMMs had $MHCII^{low}F4/80^{high}$ as well as $CD11c^+CD11b^{high}CD80^-CD64^+MerTK^+$ phenotypes. In contrast, GM-BMDCs had $MHCII^{high}F4/80^{low}$ and $CD11c^{high}CD8{\alpha}^-CD11b^+CD80^+CD64^-MerTK^{low}$ phenotypes. Interestingly, the GM-BMM population increased but GM-BMDCs decreased in a GM-CSF dose-dependent manner. Functionally, GM-BMMs showed extremely high phagocytic abilities and produced higher IL-10 upon LPS stimulation. GM-BMDCs, however, could not phagocytose as well, but were efficient at producing $TNF{\alpha}$, $IL-1{\beta}$, IL-12p70 and IL-6 as well as inducing T cell proliferation. Finally, whole transcriptome analysis revealed that GM-BMMs and GM-BMDCs are overlap with in vivo resident macrophages and dendritic cells, respectively. Taken together, our study shows the heterogeneicity of GM-CSF derived cell populations, and specifically characterizes GM-CSF derived macrophages compared to dendritic cells.

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References

  1. Banchereau, J., and Steinman, R.M. (1998). Dendritic cells and the control of immunity. Nature 392, 245-252. https://doi.org/10.1038/32588
  2. Becker, L., Liu, N.C., Averill, M.M., Yuan, W., Pamir, N., Peng, Y., Irwin, A.D., Fu, X., Bornfeldt, K.E., and Heinecke, J.W. (2012). Unique proteomic signatures distinguish macrophages and dendritic cells. PLoS One 7, e33297. https://doi.org/10.1371/journal.pone.0033297
  3. Bhattacharya, P., Gopisetty, A., Ganesh, B.B., Sheng, J.R., and Prabhakar, B.S. (2011). GM-CSF-induced, bone-marrow-derived dendritic cells can expand natural Tregs and induce adaptive Tregs by different mechanisms. J. Leukocyte Biol. 89, 235-249. https://doi.org/10.1189/jlb.0310154
  4. Cebon, J., Layton, J.E., Maher, D., and Morstyn, G. (1994). Endogenous haemopoietic growth factors in neutropenia and infection. Br. J. Haematol. 86, 265-274. https://doi.org/10.1111/j.1365-2141.1994.tb04725.x
  5. Cheers, C., Haigh, A.M., Kelso, A., Metcalf, D., Stanley, E.R., and Young, A.M. (1988). Production of colony-stimulating factors (CSFs) during infection: separate determinations of macrophage-, granulocyte-, granulocyte-macrophage-, and multi-CSFs. Infect. Immun. 56, 247-251.
  6. Chung, S., Ranjan, R., Lee, Y.G., Park, G.Y., Karpurapu, M., Deng, J., Xiao, L., Kim, J.Y., Unterman, T.G., and Christman, J.W. (2015). Distinct role of FoxO1 in M-CSF- and GM-CSF-differentiated macrophages contributes LPS-mediated IL-10: implication in hyperglycemia. J. Leukocyte Biol. 97, 327-339. https://doi.org/10.1189/jlb.3A0514-251R
  7. Crozat, K., Guiton, R., Guilliams, M., Henri, S., Baranek, T., Schwartz-Cornil, I., Malissen, B., and Dalod, M. (2010). Comparative genomics as a tool to reveal functional equivalences between human and mouse dendritic cell subsets. Immunol. Rev. 234, 177-198. https://doi.org/10.1111/j.0105-2896.2009.00868.x
  8. Egawa, M., Mukai, K., Yoshikawa, S., Iki, M., Mukaida, N., Kawano, Y., Minegishi, Y., and Karasuyama, H. (2013). Inflammatory monocytes recruited to allergic skin acquire an anti-inflammatory M2 phenotype via basophil-derived interleukin-4. Immunity 38, 570-580. https://doi.org/10.1016/j.immuni.2012.11.014
  9. Fleetwood, A.J., Lawrence, T., Hamilton, J.A., and Cook, A.D. (2007). Granulocyte-macrophage colony-stimulating factor (CSF) and macrophage CSF-dependent macrophage phenotypes display differences in cytokine profiles and transcription factor activities: implications for CSF blockade in inflammation. J. Immunol. 178, 5245-5252. https://doi.org/10.4049/jimmunol.178.8.5245
  10. Ganesh, B.B., Cheatem, D.M., Sheng, J.R., Vasu, C., and Prabhakar, B.S. (2009). GM-CSF-induced CD11c+CD8a--dendritic cells facilitate Foxp3+ and IL-10+ regulatory T cell expansion resulting in suppression of autoimmune thyroiditis. Int. Immunol. 21, 269-282. https://doi.org/10.1093/intimm/dxn147
  11. Gautier, E.L., Shay, T., Miller, J., Greter, M., Jakubzick, C., Ivanov, S., Helft, J., Chow, A., Elpek, K.G., Gordonov, S., et al. (2012). Gene-expression profiles and transcriptional regulatory pathways that underlie the identity and diversity of mouse tissue macrophages. Nat. Immunol. 13, 1118-1128. https://doi.org/10.1038/ni.2419
  12. Hashimoto, D., Miller, J., and Merad, M. (2011). Dendritic cell and macrophage heterogeneity in vivo. Immunity 35, 323-335. https://doi.org/10.1016/j.immuni.2011.09.007
  13. Helft, J., Bottcher, J., Chakravarty, P., Zelenay, S., Huotari, J., Schraml, B.U., Goubau, D., and Reis e Sousa, C. (2015). GMCSF mouse bone marrow cultures comprise a heterogeneous population of CD11c(+)MHCII(+) macrophages and dendritic cells. Immunity 42, 1197-1211. https://doi.org/10.1016/j.immuni.2015.05.018
  14. Heng, T.S., Painter, M.W., and Immunological Genome Project, C. (2008). The Immunological Genome Project: networks of gene expression in immune cells. Nat. Immunol. 9, 1091-1094. https://doi.org/10.1038/ni1008-1091
  15. Hercus, T.R., Thomas, D., Guthridge, M.A., Ekert, P.G., King-Scott, J., Parker, M.W., and Lopez, A.F. (2009). The granulocytemacrophage colony-stimulating factor receptor: linking its structure to cell signaling and its role in disease. Blood 114, 1289-1298. https://doi.org/10.1182/blood-2008-12-164004
  16. Inaba, K., Inaba, M., Deguchi, M., Hagi, K., Yasumizu, R., Ikehara, S., Muramatsu, S., and Steinman, R.M. (1993). Granulocytes, macrophages, and dendritic cells arise from a common major histocompatibility complex class II-negative progenitor in mouse bone marrow. Proc. Natl. Acad. Sci. USA 90, 3038-3042. https://doi.org/10.1073/pnas.90.7.3038
  17. Mellman, I., and Steinman, R.M. (2001). Dendritic cells: specialized and regulated antigen processing machines. Cell 106, 255-258. https://doi.org/10.1016/S0092-8674(01)00449-4
  18. Miller, J.C., Brown, B.D., Shay, T., Gautier, E.L., Jojic, V., Cohain, A., Pandey, G., Leboeuf, M., Elpek, K.G., Helft, J., et al. (2012). Deciphering the transcriptional network of the dendritic cell lineage. Nat. Immunol. 13, 888-899. https://doi.org/10.1038/ni.2370
  19. Murray, P.J., Allen, J.E., Biswas, S.K., Fisher, E.A., Gilroy, D.W., Goerdt, S., Gordon, S., Hamilton, J.A., Ivashkiv, L.B., Lawrence, T., et al. (2014). Macrophage activation and polarization: nomenclature and experimental guidelines. Immunity 41, 14-20. https://doi.org/10.1016/j.immuni.2014.06.008
  20. Nikolic, T., de Bruijn, M.F., Lutz, M.B., and Leenen, P.J. (2003). Developmental stages of myeloid dendritic cells in mouse bone marrow. Int. Immunol. 15, 515-524. https://doi.org/10.1093/intimm/dxg050
  21. Paine, R., 3rd, Morris, S.B., Jin, H., Wilcoxen, S.E., Phare, S.M., Moore, B.B., Coffey, M.J., and Toews, G.B. (2001). Impaired functional activity of alveolar macrophages from GM-CSF-deficient mice. Am. J. Physiol. Lung Cell. Mol. Physiol. 281, L1210-1218. https://doi.org/10.1152/ajplung.2001.281.5.L1210
  22. Robbins, S.H., Walzer, T., Dembele, D., Thibault, C., Defays, A., Bessou, G., Xu, H., Vivier, E., Sellars, M., Pierre, P., et al. (2008). Novel insights into the relationships between dendritic cell subsets in human and mouse revealed by genome-wide expression profiling. Genome Biol. 9, R17. https://doi.org/10.1186/gb-2008-9-1-r17
  23. Saraiva, M., and O'Garra, A. (2010). The regulation of IL-10 production by immune cells. Nat. Rev. Immunol. 10, 170-181. https://doi.org/10.1038/nri2711
  24. Satpathy, A.T., Wu, X., Albring, J.C., and Murphy, K.M. (2012). Re(de)fining the dendritic cell lineage. Nat. Immunol. 13, 1145-1154. https://doi.org/10.1038/ni.2467
  25. Seok, S.H., Heo, J.I., Hwang, J.H., Na, Y.R., Yun, J.H., Lee, E.H., Park, J.W., and Cho, C.H. (2013). Angiopoietin-1 elicits pro-inflammatory responses in monocytes and differentiating macrophages. Mol. Cells 35, 550-556. https://doi.org/10.1007/s10059-013-0088-8
  26. Xu, Y., Zhan, Y., Lew, A.M., Naik, S.H., and Kershaw, M.H. (2007). Differential development of murine dendritic cells by GM-CSF versus Flt3 ligand has implications for inflammation and trafficking. J. Immunol. 179, 7577-7584. https://doi.org/10.4049/jimmunol.179.11.7577
  27. Zhang, Y., Harada, A., Wang, J.B., Zhang, Y.Y., Hashimoto, S., Naito, M., and Matsushima, K. (1998). Bifurcated dendritic cell differentiation in vitro from murine lineage phenotype-negative c-kit+ bone marrow hematopoietic progenitor cells. Blood 92, 118-128.

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