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Published on: November 8, 2011
Tonsil stromal-cell lines expressing FDC-like properties: isolation, characterization, and interaction with B
G Skibinski1, A Skibinska, M Deckers
1Department of Surgery, University of Edinburgh, Royal Infirmary, Scotland, UK.
Insights
Stromal cells in lymphoid organs are crucial for immune responses. These cells, when cultured, enhance B cell proliferation and immunoglobulin production, highlighting their role in humoral immunity.
Area of Science:
- Immunology
- Cell Biology
- Microenvironment Research
Background:
- Secondary lymphoid organs contain lymphoid and stromal cells essential for immune responses.
- Stromal cells significantly influence tissue architecture and function.
- Understanding stromal cell interactions is key to controlling humoral immunity.
Purpose of the Study:
- To isolate and characterize human tonsil stromal cells.
- To investigate the functional properties and cytokine production of these stromal cells.
- To determine the impact of stromal cells on B cell responses.
Main Methods:
- Isolation of stromal cells from human tonsil slices via a multistage culture and depletion process.
- Phenotypic analysis using monoclonal antibodies (HLA-DR, CD54, CD44, CD45).
- Cytokine immunoassay and functional co-culture assays with B cells.
Main Results:
- Isolated stromal cells expressed HLA-DR, CD54, and CD44, but not CD45.
- Stromal cells constitutively produced IL-6, with increased production upon TNF-alpha and IFN-gamma stimulation.
- Co-culture with B cells enhanced B cell proliferation, immunoglobulin, and IL-6 production.
Conclusions:
- Human tonsil stromal cells possess unique characteristics and functional properties.
- These stromal cells exhibit follicular dendritic cell-like traits.
- Stromal cells play a vital role in supporting B cell activation and humoral immune responses through crucial signaling.
Abstract:
The microenvironment of secondary lymphoid organs consists of two major populations of cells, the lymphoid cells and a population of stromal cells that contribute to both tissue architecture and function. Interactions of both populations are essential for the development and control of humoral immune responses. In this study, stromal-cell preparations were obtained by a multistage process. This involved culturing 300-400-microm slices of human tonsil for 6-8 days at 25 degrees C, trypsin digestion of the residual explant, followed by CD45-positive-cell depletion using magnetic beads, and a final period of culture for 4 days to remove remaining nonadherent cells. Phenotyping with a panel of monoclonal antibodies revealed that the cells express HLA-DR, CD54 (ICAM-1), CD44, but no CD45 nor a range of other markers for epithelial and endothelial cells. Immunoassays of supernatants from stromal cells revealed that IL-6 was produced constitutively, and its production was increased by treatment with TNF-alpha and IFN-gamma. In contrast IL-1, IL-2, IL-4, IL-7, IL-8, IL-10, IL-12, TNF-alpha, and IFNgamma were not produced. Functional tests showed that these cells express follicular dendritic cell-like properties. Coculturing of tonsilar B cells with stromal cells resulted in enhanced proliferation and also led to increased production of immunoglobulins and IL-6, suggesting crucial signaling between these populations.
