title: Analysis of DC Functions Using CD205-DTR Knock-In Mice.
abstract: Dendritic cells (DCs) are essential antigen-presenting cells (APCs) that consist of heterogeneous subsets, mainly classified as conventional DCs (cDCs) and plasmacytoid DCs (pDCs). CD205, an endocytic type I C-type lectin-like molecule that belongs to the mannose receptor family, is mainly expressed on CD8α(+) cDCs. However, it is unclear how CD205(+) cDCs control immune responses in vivo. To evaluate the contribution of CD205(+) cDCs to the immune system, we engineered knock-in (KI) mice that express the diphtheria toxin receptor (DTR) under the control of the Cd205 gene, which allows the selective conditional ablation of CD205(+) cDCs in vivo. Conditional ablation of CD205(+) cDCs impaired the antigen-specific priming of CD8(+) T cells to generate cytotoxic T lymphocytes (CTLs) mediated through cross presentation of soluble antigen. Upon microbial infection, CD205(+) cDCs contributed to the cross priming of CD8(+) T cells for generating antibacterial CTLs to efficiently eliminate pathogens. Here, we provide a protocol for the generation of bone marrow WT/CD205-DT chimeric mice, depletion of CD205(+) DCs and assessment of depletion efficiency, and protocols for in vivo cross presentation assay, CTL generation assay, and antibacterial immunity assay.
authors.affiliation: Division of Immunology, Department of Infectious Diseases, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake, Miyazaki, 889-1692, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Fukaya
authors.affiliation: Division of Immunology, Department of Infectious Diseases, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake, Miyazaki, 889-1692, Japan.
authors.forename: Hideaki
authors.initials: H
authors.lastname: Takagi
authors.affiliation: Division of Immunology, Department of Infectious Diseases, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake, Miyazaki, 889-1692, Japan.
authors.forename: Tomofumi
authors.initials: T
authors.lastname: Uto
authors.affiliation: Division of Immunology, Department of Infectious Diseases, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake, Miyazaki, 889-1692, Japan.
authors.forename: Keiichi
authors.initials: K
authors.lastname: Arimura
authors.affiliation: Division of Immunology, Department of Infectious Diseases, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake, Miyazaki, 889-1692, Japan. katsuaki_sato@med.miyazaki-u.ac.jp.
authors.forename: Katsuaki
authors.initials: K
authors.lastname: Sato
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics* / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D045142:Cross-Priming; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology; D055879:Gene Knock-In Techniques; D066257:Heparin-binding EGF-like Growth Factor / Q000235:genetics / Q000378:metabolism*; D037181:Lectins, C-Type / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D015778:Minor Histocompatibility Antigens / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D015703:Antigens, CD; C093569:DEC-205 receptor; D066257:Heparin-binding EGF-like Growth Factor; D037181:Lectins, C-Type; D015778:Minor Histocompatibility Antigens; D011956:Receptors, Cell Surface
keywords: Cross priming; Cytotoxic T lymphocytes; Dendritic cells; Knock-in mice; Microbial infection