Abstract
Dendritic cells (DCs) are the most professional antigen-presenting cells that are indispensable for the initiation of adaptive immune responses. DCs are heterogeneous in terms of their origin, anatomical location, cell-surface markers, and functions. Previous studies have demonstrated that there exist several groups of DCs in the lamina propria (LPDC) of gastrointestinal tract, which collectively contribute to the maintenance of gut homeostasis through the regulation of the balance between active immunity and tolerance. However, although intestinal LPDCs are attractive research target for understanding the immunological mechanisms in the gut, isolation of the LPDCs is complicated and technically difficult for unskilled people. Therefore, establishment of the method to isolate intestinal LPDCs is a major obstacle in this research. Here, we describe the methods that we have established for the isolation of primary DCs from the LP of mouse small intestine. Our isolation method provides high yield of viable LP leukocytes (LPLs) including DCs. Combination with FACS sorting allows for the selective isolation of CD103+CD8α+ DCs and CD103+CD8α− DCs from the LPLs. Furthermore, isolated LPDCs can be subjected to immunological assays, such as measurement of cytokine productions following stimulation of Toll-like receptors. Thus, our methods would be useful for studying the functions of LPDCs of mouse small intestine.
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References
Mildner A, Jung S (2014) Development and function of dendritic cell subsets. Immunity 40:642–656
Pearce EJ, Everts B (2015) Dendritic cell metabolism. Nat Rev Immunol 15:18–29
Iwasaki A (2007) Mucosal dendritic cells. Annu Rev Immunol 25:381–418
Uematsu S, Akira S (2009) Immune responses of TLR5(+) lamina propria dendritic cells in enterobacterial infection. J Gastroenterol 44:803–811
Annacker O, Coombes JL, Malmstrom V et al (2005) Essential role for CD103 in the T cell-mediated regulation of experimental colitis. J Exp Med 202:1051–1061
Jang MH, Sougawa N, Tanaka T et al (2006) CCR7 is critically important for migration of dendritic cells in intestinal lamina propria to mesenteric lymph nodes. J Immunol 176:803–810
Sun CM, Hall JA, Blank RB et al (2007) Small intestine lamina propria dendritic cells promote de novo generation of Foxp3 T reg cells via retinoic acid. J Exp Med 204:1775–1785
Coombes JL, Siddiqui KR, Arancibia-Cárcamo CV et al (2007) A functionally specialized population of mucosal CD103+ DCs induces Foxp3+ regulatory T cells via a TGF-beta and retinoic acid-dependent mechanism. J Exp Med 204:1757–1764
Iwata M, Hirakiyama A, Eshima Y et al (2004) Retinoic acid imprints gut-homing specificity on T cells. Immunity 21:527–538
Uematsu S, Jang MH, Chevrier N et al (2006) Detection of pathogenic intestinal bacteria by Toll-like receptor 5 on intestinal CD11c+ lamina propria cells. Nat Immunol 7:868–874
Uematsu S, Fujimoto K, Jang MH et al (2008) Regulation of humoral and cellular gut immunity by lamina propria dendritic cells expressing Toll-like receptor 5. Nat Immunol 9:769–776
Fujimoto K, Karuppuchamy T, Takemura N et al (2011) A new subset of CD103+ CD8alpha+ dendritic cells in the small intestine expresses TLR3, TLR7, and TLR9 and induces Th1 response and CTL activity. J Immunol 186:6287–6295
Acknowledgement
We thank for Myoung Ho Jang at Pohang University of Science and Technology for his great support in establishing the isolation methods of LPDCs of mouse small intestine.
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Takemura, N., Uematsu, S. (2016). Isolation and Functional Analysis of Lamina Propria Dendritic Cells from the Mouse Small Intestine. In: Ivanov, A. (eds) Gastrointestinal Physiology and Diseases. Methods in Molecular Biology, vol 1422. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3603-8_17
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DOI: https://doi.org/10.1007/978-1-4939-3603-8_17
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3601-4
Online ISBN: 978-1-4939-3603-8
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