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Murine respiratory tract dendritic cells: isolation, phenotyping and functional studies
Claudia J Calder1, J Liversidge, Andrew D Dick
1Division of Ophthalmology, University of Bristol, UK.
Journal of Immunological Methods
|April 22, 2004
Summary
Researchers developed a consistent method to isolate respiratory tract dendritic cells (RTDCs), crucial for immune responses. This technique preserves their immature state and antigen-presenting function for further study.
Area of Science:
- Immunology
- Cell Biology
Background:
- Respiratory tract dendritic cells (RTDCs) form a network in the nasorespiratory tract, linking innate and acquired immunity.
- RTDCs play a role in nasal mucosal tolerance induction.
- Current isolation techniques yield inconsistent RTDC phenotypes and functions.
Purpose of the Study:
- To modify existing methods for consistent RTDC isolation.
- To ensure high cell viability and enriched cell numbers.
- To facilitate further phenotype and functional studies of murine RTDCs.
Main Methods:
- Enzyme digestion and Percoll density gradient centrifugation.
- Overnight culture with granulocyte-macrophage colony-stimulating factor (GM-CSF).
- Magnetic-activated cell sorting (MACS) separation.
Main Results:
- Isolated RTDCs retained an archetypal immature dendritic cell phenotype (MHCII(low) CD40(neg) CD86(neg) CD80(neg) CD11c(low)).
- Splenic-derived DCs (SDCs) showed a similar initial phenotype but could mature in vitro.
- Both RTDCs and SDCs efficiently processed and presented antigens to T cells.
Conclusions:
- A consistent method for RTDC enrichment was developed.
- The method maintains the immature phenotype of RTDCs.
- The isolated RTDCs exhibit functional antigen-presenting capabilities.