RBP-J is required for M2 macrophage polarization in response to chitin and mediates expression of a subset of M2 genes

Abstract

Development of alternatively activated (M2) macrophage phenotypes is a complex process that is coordinately regulated by a plethora of pathways and factors. Here, we report that RBP-J, a DNA-binding protein that integrates signals from multiple pathways including the Notch pathway, is critically involved in polarization of M2 macrophages. Mice deficient in RBP-J in the myeloid compartment exhibited impaired M2 phenotypes in vivo in a chitin-induced model of M2 polarization. Consistent with the in vivo findings, M2 polarization was partially compromised in vitro in Rbpj-deficient macrophages as demonstrated by reduced expression of a subset of M2 effector molecules including arginase 1. Functionally, myeloid Rbpj deficiency impaired M2 effector functions including recruitment of eosinophils and suppression of T cell proliferation. Collectively, we have identified RBPJ as an essential regulator of differentiation and function of alternatively activated macrophages.

Keywords

macrophages / RBP-J / M2 / arginase / chitin

References

[1] Chawla A, Nguyen KD, Goh YP (2011) Macrophage-mediated inflammationin metabolic disease. Nat Rev Immunol 11:738–749

[2] El Kasmi KC, Qualls JE, Pesce JT, Smith AM, Thompson RW (2008) Toll-like receptor-induced arginase 1 in macrophages thwarts effective immunity against intracellular pathogens. Nat Immunol 9:1399–1406

[3] Franklin RA, Liao W, Sarkar A, Kim MV, Bivona MR(2014)The cellular and molecular origin of tumor-associated macrophages. Science 344:921–925

[4] Fung E, Tang SM, Canner JP, Morishige K, Arboleda-Velasquez JF (2007) Delta-like4induces notch signalingin macrophages: implications for inflammation. Circulation1 15:2948–2956

[5] Hamidi H, Gustafason D, Pellegrini M, Gasson J(2011) Identification of novel targets of CSL-dependent Notch signaling in hematopoiesis. PLoS One 6:e20022

[6] Hu X, Chung AY, Wu I, Foldi J, Chen J (2008) Integrated regulationofToll-like receptor responsesbyNotch and interferon-gamma pathways. Immunity 29:691–703

[7] Ivashkiv LB (2013) Epigenetic regulation of macrophage polarization and function. Trends Immunol 34:216–223

[8] Klinakis A, Lobry C, Abdel-Wahab O, Oh P, Haeno H(2011)A novel tumour-suppressor function for the Notch pathway in myeloid leukaemia. Nature 473:230–233

[9] Kopan R, Ilagan MX (2009) The canonical Notch signalingpathway: unfolding the activation mechanism. Cell 137:216–233

[10] Lewis KL, Caton ML, Bogunovic M, Greter M, Grajkowska LT (2011) Notch2 receptor signaling controls functional differentiation of dendritic cells in the spleen and intestine. Immunity 35:780–791

[11] Li Y, Hibbs MA, Gard AL, Shylo NA, Yun K (2012) Genome-wide analysis of N1ICD/RBPJ targets in vivo reveals direct transcriptional regulation of Wnt, SHH, and hippo pathway effectors by Notch1. Stem Cells 30(4):741–752

[12] Monsalve E, Perez MA, Rubio A, Ruiz-Hidalgo MJ, Baladron V (2006) Notch-1 up-regulation and signaling following macrophage activation modulates gene expression patterns known to affect antigen-presenting capacity and cytotoxic activity. J Immunol 176:5362–5373

[13] Monsalve E, Ruiz-Garcia A, Baladron V, Ruiz-Hidalgo MJ, Sanchez-Solana B (2009) Notch1 upregulates LPS-induced macrophage activation by increasing NF-kappaB activity. Eur J Immunol 39:2556–2570

[14] Mosser DM, Edwards JP (2008) Exploring the full spectrum of macrophage activation. Nat Rev Immunol 8:958–969

[15] Murray PJ, Wynn TA (2011) Protective and pathogenic functions of macrophage subsets. Nat Rev Immunol 11:723–737

[16] Murray PJ, Allen JE, Biswas SK, Fisher EA, Gilroy DW (2014) Macrophage activation and polarization: nomenclature and experimental guidelines. Immunity 41:14–20

[17] Odegaard JI, Ricardo-Gonzalez RR, Goforth MH, Morel CR, Subramanian V (2007) Macrophage-specific PPARgamma controls alternative activation and improves insulin resistance. Nature 447:1116–1120

[18] Outtz HH, Wu JK, Wang X, Kitajewski J (2010) Notch1 deficiency results in decreased inflammation during wound healing and regulates vascular endothelial growth factor receptor-1 and inflammatory cytokine expression in macrophages. J Immunol 185:4363–4373

[19] Palaga T, Buranaruk C, Rengpipat S, Fauq AH, Golde TE (2008) Notch signaling is activated by TLR stimulation and regulates macrophage functions. EurJ Immunol 38:174–183

[20] Palomero T, Lim WK, Odom DT, Sulis ML, Real PJ (2006) NOTCH1 directly regulates c-MYC and activates a feed-forwardloop transcriptional network promoting leukemic cell growth. Proc Natl Acad Sci USA 103:18261–18266

[21] Pesce JT, Ramalingam TR, Mentink-Kane MM, Wilson MS, El Kasmi KC (2009) Arginase-1-expressing macrophages suppress Th2 cytokine-driven inflammation and fibrosis. PLoS Pathog 5: e1000371

[22] Pourcet B, Feig JE, Vengrenyuk Y, Hobbs AJ, Kepka-Lenhart D (2011) LXRalpha regulates macrophage arginase1through PU. 1 and interferon regulatory factor 8. Circ Res 109:492–501

[23] Radtke F, Fasnacht N, Macdonald HR (2010) Notch signalingin the immune system. Immunity 32:14–27

[24] Reese TA, Liang HE, Tager AM, Luster AD, Van Rooijen N (2007) Chitin induces accumulation in tissue of innate immune cells associated with allergy. Nature 447:92–96

[25] Satoh T, Takeuchi O, Vandenbon A, Yasuda K, Tanaka Y(2010) The Jmjd3-Irf4 axis regulates M2 macrophage polarization and host responses against helminth infection. Nat Immunol 11:936–944

[26] Tsao PN, Wei SC, Huang MT, Lee MC, Chou HC (2011) Lipopolysaccharide-induced Notch signaling activation through JNK-dependent pathway regulates inflammatory response. J Biomed Sci 18:56

[27] Wang YC, He F, Feng F, Liu XW, Dong GY (2010) Notch signaling determines the M1 versus M2 polarization of macrophages in antitumor immune responses. Cancer Res 70:4840–4849

[28] Wang H, Zou J, Zhao B, Johannsen E, Ashworth T (2011) Genome-wide analysis reveals conserved and divergent features of Notch1/RBPJ binding in human and murine T-lymphoblastic leukemia cells. Proc Natl Acad Sci USA 108:14908–14913

[29] Xu H, Zhu J, Smith S, Foldi J, Zhao B (2012) Notch-RBP-J signaling regulates the transcription factor IRF8 to promote inflammatory macrophage polarization. Nat Immunol 13:642–650

[30] Yuan JS, Kousis PC, Suliman S, Visan I, Guidos CJ (2010) Functions of notch signaling in the immune system: consensus and controversies. Annu Rev Immunol 28:343–365

[31] Zhang W, Xu W, Xiong S (2010) Blockade of Notch1 signaling alleviates murine lupus via blunting macrophage activation and M2b polarization. J Immunol 184:6465–6478

[32] Zhang Q, Wang C, Liu Z, Liu X, Han C (2012) Notch signal suppresses TLR-triggered inflammatory responses in macrophages by inhibiting ERK1/2-mediated NF-kappaB activation. J Biol Chem 28:jbc-M111

[33] Zhao B, Zou J, Wang H, Johannsen E, Peng CW (2011) Epstein-Barr virus exploits intrinsic B-lymphocyte transcription programs to achieve immortal cell growth. Proc Natl Acad Sci USA 108:14902–14907

[34] Zhao B, Grimes SN, Li S, Hu X, Ivashkiv LB (2012) TNF-induced osteoclastogenesis and inflammatory bone resorption are inhibitedby transcription factor RBP-J. J Exp Med 209:319–334

RIGHTS & PERMISSIONS

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.