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An Efficient and High Yield Method for Isolation of Mouse Dendritic Cell Subsets
Published on: April 18, 2016
Characterization of accessory molecules in murine lung dendritic cell function: roles for CD80, CD86, CD54, and CD40L
B J Masten1, J L Yates, A M Pollard Koga
1Department of Pathology, University of New Mexico, Albuquerque 87131-5301, USA.
Insights
Murine lung dendritic cells (DCs) stimulate T cell responses. Interstitial macrophages enhance DC function, and costimulatory molecules CD80 and CD86 are crucial for T cell proliferation.
Area of Science:
- Immunology
- Cell Biology
Background:
- Dendritic cells (DCs) are key antigen-presenting cells that initiate immune responses.
- The role of lung DCs and their interaction with other lung cells in immune modulation is not fully understood.
Purpose of the Study:
- To investigate the accessory properties and stimulatory function of murine lung dendritic cells (DCs).
- To analyze the expression of cell surface molecules on lung DCs.
- To determine the role of costimulatory molecules in DC-mediated T cell proliferation.
Main Methods:
- Enrichment of lung DCs using fluorescence-activated cell sorting.
- Analysis of cell surface molecule expression.
- Assessment of MLR (mixed leukocyte reaction) to evaluate T cell proliferation and accessory function.
Main Results:
- Lung DCs exhibited potent accessory properties in vitro, stimulating T cell responses.
- Interstitial macrophages (IM) enhanced DC-initiated T cell proliferation via cell contact.
- Lung DCs expressed typical DC surface molecules, including CD11a, CD54, CD80, and CD86.
- Costimulatory molecules CD80 and CD86 were critical for T cell proliferation, with CD80 providing a signal independently of CD86.
Conclusions:
- Murine lung DCs are potent stimulators of primary T cell responses.
- Local lung cells, like IM, can influence DC function.
- CD80 plays a significant role in costimulation of allogeneic T cells, even when CD86 signaling is absent or impaired.
Abstract:
Lung dendritic cells (DCs) from mice were enriched to 92-99% purity using a multistep enrichment protocol which included fluorescence-activated cell sorting. DCs were analyzed for expression of cell surface molecules, function in a mixed leukocyte reaction (MLR), and dependence on accessory molecules in stimulating an MLR. DCs possessed potent accessory properties in vitro, while interstitial macrophages (IM), which are Ia-negative, displayed little MLR-stimulating function of their own. However, IM were capable of enhancing DC-initiated T cell proliferation via a cell contact mechanism. These results indicated that murine lung DCs functioned as stimulators of primary T cell responses, and that cells in the local environment influenced their function. Lung DCs expressed surface molecules typical of DCs from other sites including CD11a, CD54, CD80, and CD86. As is true for DCs in other sites, costimulatory molecules including CD80, CD86, CD40L, CD2, CD54, and CD11a played important roles in lung DC-initiated T cell proliferation. Interestingly, anti-CD86 monoclonal antibody (mAb) had little inhibitory effect on the MLR unless it was added in combination with anti-CD80 mAb. These studies suggest that CD80 on lung DCs can provide a costimulatory signal to allogeneic T cells in the absence of CD86 signaling, but that CD86 functions poorly except when CD80 is also engaged.
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