Abstract
C-type lectins bind to carbohydrate structures in a Ca2+-dependent manner. Some transmembrane forms of lectins act as innate immune receptors and induce signal transduction pathways in macrophages and dendritic cells (DCs). Expressing these receptors in cells bearing a reporter gene is a useful tool to investigate ligand binding and recognition. However, it cannot be used to quantify the precise affinity of the interaction, and the involvement of other proteins remains a possibility. Direct binding between a receptor and its ligand can be investigated using an immunoglobulin receptor (Ig)-fused soluble protein. This binding can be assessed using enzyme-linked immunosorbent assays and flow cytometry, and the fusion protein may also be used in a glycan array. In this chapter, we explain the generation of Ig fusion proteins and subsequent binding assays using these proteins.
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References
van Kooyk Y, Geijtenbeek TB (2003) DC-SIGN: escape mechanism for pathogens. Nat Rev Immunol 3:697–709
Osorio F, Reis e Sousa C (2011) Myeloid C-type lectin receptors in pathogen recognition and host defense. Immunity 34:651–664
Rogers NC, Slack EC, Edwards AD et al (2005) Syk-dependent cytokine induction by Dectin-1 reveals a novel pattern recognition pathway for C type lectins. Immunity 22:507–517
Saijo S, Ikeda S, Yamabe K et al (2010) Dectin-2 recognition of alpha-mannans and induction of Th17 cell differentiation is essential for host defense against Candida albicans. Immunity 32:681–691
Yonekawa A, Saijo S, Hoshino Y et al (2014) Dectin-2 is a direct receptor for mannose-capped lipoarabinomannan of mycobacteria. Immunity 41:402–413
Yamasaki S, Ishikawa E, Sakuma M et al (2008) Mincle is an ITAM-coupled activating receptor that senses damaged cells. Nat Immunol 9:1179–1188
Ishikawa E, Ishikawa T, Morita YS et al (2009) Direct recognition of the mycobacterial glycolipid, trehalose dimycolate, by C-type lectin Mincle. J Exp Med 206:2879–2888
Kanazawa N, Tashiro K, Inaba K et al (2003) Dendritic cell immunoactivating receptor, a novel C-type lectin immunoreceptor, acts as an activating receptor through association with Fc receptor gamma chain. J Biol Chem 278:32645–32652
Toyonaga K, Torigoe S, Motomura Y et al (2016) C-type Lectin receptor DCAR recognizes mycobacterial Phosphatidyl-inositol Mannosides to promote a Th1 response during infection. Immunity 45:1245–1257
Bakker AB, Baker E, Sutherland GR et al (1999) Myeloid DAP12-associating lectin (MDL)-1 is a cell surface receptor involved in the activation of myeloid cells. Proc Natl Acad Sci U S A 96:9792–9796
Chen ST, Lin YL, Huang MT et al (2008) CLEC5A is critical for dengue-virus-induced lethal disease. Nature 453:672–676
Nagata M, Izumi Y, Ishikawa E et al (2017) Intracellular metabolite β-glucosylceramide is an endogenous Mincle ligand possessing immunostimulatory activity. Proc Natl Acad Sci U S A 114:E3285–E3294
Mayer S, Moeller R, Monteiro JT et al (2018) C-type Lectin receptor (CLR)-fc fusion proteins as tools to screen for novel CLR/bacteria interactions: an exemplary study on preselected. Front Immunol 9:213
Maglinao M, Eriksson M, Schlegel MK et al (2014) A platform to screen for C-type lectin receptor-binding carbohydrates and their potential for cell-specific targeting and immune modulation. J Control Release 175:36–42
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Watanabe, M., Omahdi, Z., Yamasaki, S. (2020). Direct Binding Analysis Between C-Type Lectins and Glycans Using Immunoglobulin Receptor Fusion Proteins. In: Hirabayashi, J. (eds) Lectin Purification and Analysis. Methods in Molecular Biology, vol 2132. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0430-4_12
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DOI: https://doi.org/10.1007/978-1-0716-0430-4_12
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Publisher Name: Humana, New York, NY
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