Abstract
Dendritic cells (DCs) are central to initiate antigen-specific immunity and tolerance. The in vivo development and distribution of DCs are now better understood even in nonlymphoid tissues [1]. Atherosclerosis is a chronic inflammatory disease of blood vessels and DCs are highly enriched in the intimal area of the aorta, which is predisposed to develop atherosclerosis. Previously, we were the first to show antigen presenting DCs and their subsets in the aorta [2, 3]. Here, we discuss several useful methods to characterize not only DCs but also other immune cells in steady state and atherosclerotic aorta. These comprise multiparameter flow cytometry strategies including intracellular staining and cell sorting, en face immunohistochemistry of DCs and regulatory T cells (Tregs), and Oil Red O staining of atherosclerotic lesions in the aorta.
The original version of this chapter was revised. The erratum to this chapter is available at: DOI 10.1007/978-1-4939-6786-5_31
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References
Steinman RM (2012) Decisions about dendritic cells: past, present, and future. Annu Rev Immunol 30:1–22
Choi JH, Do Y, Cheong C, Koh H, Boscardin SB, Oh YS, Bozzacco L, Trumpfheller C, Park CG, Steinman RM (2009) Identification of antigen-presenting dendritic cells in mouse aorta and cardiac valves. J Exp Med 206:497–505
Choi JH, Cheong C, Dandamudi DB, Park CG, Rodriguez A, Mehandru S, Velinzon K, Jung IH, Yoo JY, Oh GT, Steinman RM (2011) Flt3 signaling-dependent dendritic cells protect against atherosclerosis. Immunity 35:819–831
Paulson KE, Zhu SN, Chen M, Nurmohamed S, Jongstra-Bilen J, Cybulsky MI (2010) Resident intimal dendritic cells accumulate lipid and contribute to the initiation of atherosclerosis. Circ Res 106:383–390
Gautier EL, Huby T, Saint-Charles F, Ouzilleau B, Pirault J, Deswaerte V, Ginhoux F, Miller ER, Witztum JL, Chapman MJ, Lesnik P (2009) Conventional dendritic cells at the crossroads between immunity and cholesterol homeostasis in atherosclerosis. Circulation 119:2367–2375
Buono C, Pang H, Uchida Y, Libby P, Sharpe AH, Lichtman AH (2004) B7-1/B7-2 costimulation regulates plaque antigen-specific T-cell responses and atherogenesis in low-density lipoprotein receptor-deficient mice. Circulation 109:2009–2015
Sun J, Hartvigsen K, Chou MY, Zhang Y, Sukhova GK, Zhang J, Lopez-Ilasaca M, Diehl CJ, Yakov N, Harats D, George J, Witztum JL, Libby P, Ploegh H, Shi GP (2010) Deficiency of antigen-presenting cell invariant chain reduces atherosclerosis in mice. Circulation 122:808–820
Subramanian M, Thorp E, Hansson GK, Tabas I (2013) Treg-mediated suppression of atherosclerosis requires MYD88 signaling in DCs. J Clin Invest 123:179–188
Weber C, Meiler S, Doring Y, Koch M, Drechsler M, Megens RT, Rowinska Z, Bidzhekov K, Fecher C, Ribechini E, van Zandvoort MA, Binder CJ, Jelinek I, Hristov M, Boon L, Jung S, Korn T, Lutz MB, Forster I, Zenke M, Hieronymus T, Junt T, Zernecke A (2011) CCL17-expressing dendritic cells drive atherosclerosis by restraining regulatory T cell homeostasis in mice. J Clin Invest 121:2898–2910
Ait-Oufella H, Salomon BL, Potteaux S, Robertson AK, Gourdy P, Zoll J, Merval R, Esposito B, Cohen JL, Fisson S, Flavell RA, Hansson GK, Klatzmann D, Tedgui A, Mallat Z (2006) Natural regulatory T cells control the development of atherosclerosis in mice. Nat Med 12:178–180
Ishibashi S, Brown MS, Goldstein JL, Gerard RD, Hammer RE, Herz J (1993) Hypercholesterolemia in low density lipoprotein receptor knockout mice and its reversal by adenovirus-mediated gene delivery. J Clin Invest 92:883–893
Zhang SH, Reddick RL, Piedrahita JA, Maeda N (1992) Spontaneous hypercholesterolemia and arterial lesions in mice lacking apolipoprotein E. Science 258:468–471
Butcher MJ, Herre M, Ley K, Galkina E (2011) Flow cytometry analysis of immune cells within murine aortas. J Vis Exp pii:2848
Gautier EL, Shay T, Miller J, Greter M, Jakubzick C, Ivanov S, Helft J, Chow A, Elpek KG, Gordonov S, Mazloom AR, Ma’ayan A, Chua WJ, Hansen TH, Turley SJ, Merad M, Randolph GJ (2012) Gene-expression profiles and transcriptional regulatory pathways that underlie the identity and diversity of mouse tissue macrophages. Nat Immunol 13:1118–1128
Jakubzick C, Gautier EL, Gibbings SL, Sojka DK, Schlitzer A, Johnson TE, Ivanov S, Duan Q, Bala S, Condon T, van Rooijen N, Grainger JR, Belkaid Y, Ma’ayan A, Riches DW, Yokoyama WM, Ginhoux F, Henson PM, Randolph GJ (2013) Minimal differentiation of classical monocytes as they survey steady-state tissues and transport antigen to lymph nodes. Immunity 39:599–610
Cheong C, Choi JH (2012) Dendritic cells and regulatory T cells in atherosclerosis. Mol Cells 34:341–347
Paigen B, Morrow A, Holmes PA, Mitchell D, Williams RA (1987) Quantitative assessment of atherosclerotic lesions in mice. Atherosclerosis 68:231–240
Acknowledgment
This work was supported by grants from National Research Foundation of Korea (GRN-2013S1A2A2035348) (JHC and CC) and Canadian Institutes of Health Research (CIHR, MOP-125933 and HIG-133050) (CC). CC holds a Chercheur-Boursier Junior of Fonds de recherche du Québec-Santé (FRQS)—Société québécoise d’hypertension artérielle (SQHA) and CIHR New Investigator Salary Award.
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Yun, T.J., Lee, J.S., Shim, D., Choi, JH., Cheong, C. (2017). Isolation and Characterization of Aortic Dendritic Cells and Lymphocytes in Atherosclerosis. In: Clausen, B., Laman, J. (eds) Inflammation. Methods in Molecular Biology, vol 1559. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6786-5_29
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DOI: https://doi.org/10.1007/978-1-4939-6786-5_29
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