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Updated: Mar 21, 2026
Development and Functional Characterization of Murine Tolerogenic Dendritic Cells
Published on: May 18, 2018
Analysis of DC Functions Using CD205-DTR Knock-In Mice
Tomohiro Fukaya1, Hideaki Takagi1, Tomofumi Uto1
1Division of Immunology, Department of Infectious Diseases, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake, Miyazaki, 889-1692, Japan.
Insights
CD205-expressing dendritic cells (DCs) are crucial for initiating CD8+ T cell responses. Depleting these cells impairs the generation of cytotoxic T lymphocytes (CTLs) needed to fight infections.
Area of Science:
- Immunology
- Cell Biology
Background:
- Dendritic cells (DCs) are key antigen-presenting cells (APCs) with diverse subsets, including conventional DCs (cDCs) and plasmacytoid DCs (pDCs).
- CD205, a C-type lectin, is primarily found on CD8α(+) cDCs, but its role in immune regulation in vivo remains largely unknown.
Purpose of the Study:
- To investigate the in vivo function of CD205-expressing conventional DCs (CD205+ cDCs) in adaptive immune responses.
- To develop a model for selective depletion of CD205+ cDCs to study their contribution to T cell priming and immunity.
Main Methods:
- Generation of knock-in (KI) mice expressing diphtheria toxin receptor (DTR) under the Cd205 gene promoter for conditional CD205+ cDC ablation.
- Utilized bone marrow chimeric mice, diphtheria toxin administration for DC depletion, and assessed depletion efficiency.
- Employed in vivo cross-presentation assays, cytotoxic T lymphocyte (CTL) generation assays, and antibacterial immunity assays.
Main Results:
- Conditional ablation of CD205+ cDCs significantly impaired antigen-specific priming of CD8+ T cells via cross-presentation of soluble antigens.
- CD205+ cDCs were essential for generating antibacterial CTLs, contributing to pathogen elimination during microbial infection.
- The study provides detailed protocols for generating chimeric mice, depleting CD205+ DCs, and evaluating their functional capacity.
Conclusions:
- CD205+ cDCs play a critical role in initiating CD8+ T cell responses through cross-presentation, particularly for generating CTLs against soluble antigens.
- These DCs are vital for effective antibacterial immunity, highlighting their importance in adaptive anti-pathogen responses.
- The developed mouse model and protocols enable further investigation into the specific functions of CD205+ cDCs in various immune contexts.
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.
