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Published on: November 4, 2016
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Isolation of Splenic Dendritic Cells Using Fluorescence-activated Cell Sorting.
Simon J Tavernier1, Fabiola Osorio2, Sophie Janssens1
1Unit immunoregulation and mucosal immunology, VIB inflammation research center, Ghent, Belgium; GROUP-ID consortium, Ghent University and University Hospital, Ghent, Belgium; Department of respiratory medicine, Ghent University, Ghent, Belgium.
Bio-Protocol
|July 5, 2016
Summary
Inositol Requiring Enzyme (IRE)-1 is active in splenic CD8a+ dendritic cells (DCs). Loss of its target, X-box binding protein (XBP)-1, impairs antigen presentation by these specific immune cells.
Area of Science:
- Immunology
- Cell Biology
Background:
- The spleen is a key immune organ with diverse dendritic cell (DC) subsets performing distinct immune functions.
- Splenic CD8a+ DCs are crucial for immune responses, but their specific molecular pathways are not fully understood.
Purpose of the Study:
- To investigate the role of Inositol Requiring Enzyme (IRE)-1 signaling in splenic CD8a+ DCs.
- To establish a protocol for isolating specific DC subsets for experimental analysis.
Main Methods:
- Utilized genetic models to study the function of IRE-1 and its downstream target, X-box binding protein (XBP)-1.
- Investigated antigen cross-presentation capabilities of splenic CD8a+ DCs in vitro.
- Developed a protocol for ex vivo isolation of splenic DC subsets.
Main Results:
- IRE-1 signaling pathway is specifically activated in splenic CD8a+ DCs.
- Loss of XBP-1 in these cells leads to impaired cross-presentation of antigens derived from dead cells.
- A reliable method for isolating specific DC subsets from the spleen was established.
Conclusions:
- IRE-1/XBP-1 pathway is essential for the function of splenic CD8a+ DCs in antigen cross-presentation.
- This study provides a valuable tool for further research into DC subset-specific immunity.
