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An Efficient and High Yield Method for Isolation of Mouse Dendritic Cell Subsets
Published on: April 18, 2016
Characterization and functional activity of dendritic cells from rat choroid
A Choudhury1, V A Pakalnis, W E Bowers
1Department of Microbiology and Immunology, University of South Carolina School of Medicine, Columbia 29208.
Insights
Functional dendritic cells (DC) were identified in the posterior eye cup, specifically within the choroid. These cells play a role in immune responses and may be involved in posterior uveitis.
Area of Science:
- Ophthalmology
- Immunology
- Cell Biology
Background:
- Dendritic cells (DC) are crucial immune cells involved in T-cell responses.
- The presence and function of DC in the posterior eye compartment are not well-established.
Purpose of the Study:
- To investigate the presence and function of dendritic cells (DC) in the posterior eye cup.
- To characterize the localization and potential role of these cells in ocular immune responses.
Main Methods:
- Cell culture of posterior eye cup preparations.
- Assays for T-cell responses and mixed leukocyte reactions (MLR).
- Immunomagnetic separation and immunofluorescent staining using anti-DC antibodies.
- Cryostat sectioning and histological analysis of eye tissue.
Main Results:
- Eye cup cell cultures exhibited accessory T-cell activity and MLR stimulatory capacity, characteristic of DC.
- These activities were associated with low-density cells.
- Immunoselection identified a distinct subpopulation of low-density cells with DC morphology and surface antigens.
- These cells were localized to the choroid, distinct from the sclera and retina.
Conclusions:
- Functional dendritic cells (DC) are present in the choroid of the eye.
- These choroidal DC may play a significant role in the inflammatory processes of posterior uveitis.
Abstract:
Cell preparations from the posterior eye cup of the eye cultured for 2 days exhibited accessory activity for T-cell responses to a mitogenic treatment and stimulatory activity in a mixed leukocyte reaction (MLR), two functions characteristic of dendritic cells (DC). These activities both partitioned with cells having a low buoyant density, another characteristic of DC. Immunomagnetic separations with monoclonal antibodies against lymphoid dendritic cell surface antigens revealed that the accessory activity of the low-density cells was entirely associated with a small population of cells positively selected by these antibodies. Immunofluorescent staining with these same antibodies also revealed a small subpopulation of low-density cells having the morphology of DC. On cryostat sections of eye tissue the positively-stained cells were localized in the choroid and were not observed within the sclera or the retina. Based on these results we conclude that there are functional DC in the choroid, and we speculate that they may have a significant role in the inflammatory process during posterior uveitis.
