In vivo and in vitro analyses of α-galactosylceramide uptake by conventional dendritic cell subsets using its fluorescence-labeled derivative.
Conventional dendritic cells (cDCs) present α-galactosylceramide (αGC) to invariant natural killer T (iNKT) cells through CD1d. Among cDC subsets, CD8(+) DCs efficiently induce IFN-γ production in iNKT cells. Using fluorescence-labeled αGC, we showed that CD8(+) DCs incorporated larger amounts of αGC and kept it intact longer than CD8(-) DCs. Histological analyses revealed that Langerin(+)CD8(+) DCs in the splenic marginal zone, which was the unique equipment to capture blood-borne antigens, preferably incorporated αGC, and the depletion of Langerin(+) cells decreased IFN-γ and IL-12 production in response to αGC. Furthermore, splenic Langerin(+)CD8(+) DCs expressed more membrane-bound CXCL16, which possibly anchored iNKT cells in the marginal zone, than CD8(-) DCs. Collectively, it is suggested that the cellular properties and localization of CD8(+) DCs are important for stimulation of iNKT cells by αGC.
Department of Animal Development and Physiology, Graduate School of Biostudies, Kyoto University, Yoshida-Konoe, Sakyo, Kyoto 606-8501, Japan.
Maki
M
Ushida
Department of Animal Development and Physiology, Graduate School of Biostudies, Kyoto University, Yoshida-Konoe, Sakyo, Kyoto 606-8501, Japan.
Tomonori
T
Iyoda
Department of Animal Development and Physiology, Graduate School of Biostudies, Kyoto University, Yoshida-Konoe, Sakyo, Kyoto 606-8501, Japan.
Mitsuhiro
M
Kanamori
Center for Stem Cell Biology and Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato, Tokyo 108-8639, Japan.
Hiroshi
H
Watarai
Department of Animal Development and Physiology, Graduate School of Biostudies, Kyoto University, Yoshida-Konoe, Sakyo, Kyoto 606-8501, Japan. Electronic address: ktakahar@zoo.zool.kyoto-u.ac.jp.
Kazuhiko
K
Takahara
Department of Animal Development and Physiology, Graduate School of Biostudies, Kyoto University, Yoshida-Konoe, Sakyo, Kyoto 606-8501, Japan.
Kayo
K
Inaba
D000818:Animals; D000954:Antigens, Surface / Q000276:immunology / Q000378:metabolism; D016827:CD8 Antigens / Q000276:immunology / Q000378:metabolism; D002232:Carbocyanines / Q000737:chemistry; D000075743:Chemokine CXCL16; D054427:Chemokine CXCL6 / Q000276:immunology / Q000378:metabolism; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D005434:Flow Cytometry; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry; D005699:Galactosylceramides / Q000276:immunology* / Q000378:metabolism / Q000493:pharmacokinetics; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D018664:Interleukin-12 / Q000276:immunology / Q000378:metabolism; D037181:Lectins, C-Type / Q000276:immunology / Q000378:metabolism; D037241:Mannose-Binding Lectins / Q000276:immunology / Q000378:metabolism; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008856:Microscopy, Fluorescence; D055611:Natural Killer T-Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D013154:Spleen / Q000166:cytology / Q000276:immunology / Q000378:metabolism
D000954:Antigens, Surface; D016827:CD8 Antigens; D002232:Carbocyanines; C498497:Cd207 protein, mouse; D000075743:Chemokine CXCL16; D054427:Chemokine CXCL6; C498926:Cxcl16 protein, mouse; D005456:Fluorescent Dyes; D005699:Galactosylceramides; D037181:Lectins, C-Type; D037241:Mannose-Binding Lectins; C493154:alpha-galactosylceramide; C085321:cyanine dye 5; D018664:Interleukin-12; D007371:Interferon-gamma
Dendritic cells; IFN-γ; Subsets; iNKT cells; α-Galactosylceramide