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Updated: Aug 11, 2026
An Efficient and High Yield Method for Isolation of Mouse Dendritic Cell Subsets
Published on: April 18, 2016
Identification of a CD8alpha(+) dendritic cell subpopulation in rat spleen and evaluation of its OX-62 expression
M de la Mata1, C M Riera, P Iribarren
1Immunology, National University of Córdoba, Córdoba, Argentina.
Insights
Researchers identified distinct dendritic cell (DC) subpopulations in rat spleens. CD8alpha(-) spleen DCs were more effective at initiating immune responses compared to CD8alpha(+) DCs.
Area of Science:
- Immunology
- Cell Biology
- Dendritic Cell Research
Background:
- Dendritic cells (DCs) are crucial antigen-presenting cells in the immune system.
- Spleen and bone marrow are primary sources for DC isolation and study.
- Characterization of DC subpopulations is essential for understanding immune regulation.
Purpose of the Study:
- To characterize dendritic cell (DC) subpopulations in rat spleen.
- To investigate the immune response induction capacity of different spleen DC subsets.
- To explore the role of CD8alpha and OX-62 markers in DC heterogeneity.
Main Methods:
- Isolation and characterization of rat spleen and bone marrow-derived DCs.
- Flow cytometry analysis for CD8alpha and OX-62 expression.
- Magnetic-activated cell sorting (MACS) to separate DC subpopulations.
- In vivo primary immune response induction assay using Ovalbumin (OVA).
Main Results:
- A CD8alpha(+) DC subpopulation constituted 19-48% of total spleen DCs.
- OX-62 expression was observed on 41-59% of spleen DCs, with coexpression on some CD8alpha(+) DCs.
- Both CD8alpha(+) and CD8alpha(-) spleen DC subpopulations induced in vivo immune responses.
- CD8alpha(-) spleen DCs induced a significantly higher immune response than CD8alpha(+) DCs.
- Bone marrow-derived DCs were negative for CD8alpha and OX-62.
Conclusions:
- Rat spleen contains distinct DC subpopulations, including CD8alpha(+) and CD8alpha(-).
- CD8alpha(-) spleen DCs are more potent inducers of primary in vivo immune responses.
- DC preparations may contain subpopulations with different lineages, indicated by OX-62 expression.
Abstract:
Rat spleen DC and bone marrow-derived DC were isolated and characterized by morphology and flow cytometry. We found a CD8alpha(+) DC subpopulation representing 19-48% (27.4 +/- 12.0) of total spleen DC. The OX-62 expression on total spleen DC was 41-59% (51.8 +/- 7.5). Myeloid bone marrow-derived DC were negative for CD8alpha and OX-62. We demonstrated the coexpression of CD8alpha and OX-62 molecules, at least in a portion CD8alpha(+) spleen DC. Both CD8alpha(+) and CD8alpha(-) spleen DC subpopulations separated by MACS were able to induce an in vivo primary immune response to OVA. The immune response induced by the CD8alpha(-) DC subpopulation was higher (P < 0.05). We identified a CD8alpha(+) DC subpopulation in rat spleen less effective in inducing an immune response than CD8alpha(-) DC. Moreover, our results suggest the presence of DC subpopulations with different lineages in DC preparations based on OX-62 expression.
