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Isolation, Processing and Analysis of Murine Gingival Cells
Published on: July 2, 2013
Differential expression of costimulatory molecules in chronic inflammatory periodontal disease tissue
K Orima1, K Yamazaki, T Aoyagi
1Department of Periodontology, Niigata University School of Dentistry, Niigata, Japan.
Insights
This study investigated costimulatory molecules in periodontal disease, finding variable expression of CD40 ligand and CTLA-4 on T cells in gingival tissues, suggesting a role in regulating B cell activation.
Area of Science:
- Immunology
- Periodontology
Background:
- Chronic inflammatory periodontal disease involves B cell activation and immunoglobulin production.
- In situ expression of costimulatory molecules in humoral immunity during periodontitis is not well understood.
Purpose of the Study:
- To investigate the in situ expression of key costimulatory molecules (CD40, CD40L, CD80, CD86, CD28, CTLA-4) on lymphocytes in periodontal disease.
- To analyze the in vitro kinetics of T cell and B cell activation markers.
Main Methods:
- Immunohistochemistry on gingival tissue from periodontitis patients.
- Flow cytometry analysis of peripheral blood T and B cells.
- Quantification of CD40L, CD28, CTLA-4, CD40, CD80, and CD86 expression.
Main Results:
- In vitro T cell activation upregulated CD40L and CTLA-4; B cell activation upregulated CD80 and CD86.
- Gingival tissues showed high expression of CD28 on T cells and CD80/CD86 on B cells.
- CD40L and CTLA-4 expression in gingival tissues was low and highly variable, with reciprocal patterns observed.
Conclusions:
- Variable CD40L and CTLA-4 expression in periodontal lesions may indicate dynamic immunological activity.
- These molecules likely participate in the regulation of B cell activation within the periodontal disease microenvironment.
Abstract:
Although B cell activation and subsequent immunoglobulin production are the immunopathological features of chronic inflammatory periodontal disease, in situ expression of costimulatory molecules in humoral immunity has not been investigated. In the present study we examined the expression of CD40, CD40 ligand (CD40L), CD80, CD86, CD28 and cytolytic T lymphocyte-associated antigen-4 (CTLA-4) on lymphocytes immunohistochemically. Cryostat sections were prepared from the gingival tissue samples of 14 patients with moderate to advanced adult periodontitis. In vitro kinetics of the expression of CD40L and CTLA-4 by peripheral blood T cells and that of CD80 and CD86 by peripheral blood B cells were also investigated by flow cytometry. Positive percentage expression of CD40L, CD28 and CTLA-4, and CD40, CD80 and CD86 was calculated for the number of CD3+ and CD19+ cells, respectively. Flow cytometric analysis demonstrated that the expression of CD40L and CTLA-4 on T cells, and CD80 and CD86 on B cells of peripheral blood was up-regulated upon activation. While most T cells and B cells expressed CD28, and CD80 and CD86, respectively, in gingival tissues, the expression of CD40L and CTLA-4 was lower but highly variable between specimens. Furthermore, these two molecules seemed to be expressed reciprocally in the lesion. As both CD40L and CTLA-4 expression are induced transiently by stimulation, variability in the expression of the molecules may reflect immunological activities and participation in the regulation of B cell activation of the lesion.
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