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Multiple Dendritic Cell (DC) Subpopulations in Human Gingiva and Association of Mature DCs with CD4+ T-cells in situ
R. Jotwani and
C.W. Cutler*
Department of Periodontics, 110 Rockland Hall, School of Dental Medicine, State University of New York at Stony Brook, Stony Brook, NY 11794-8703;
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Figure 1. Distinct tissue compartmentalization of Langerhans cells and dermal DCs in chronic periodontitis (CP). Shown are double-immunofluorescence stainings of representative CP gingival tissue. (A) Clear compartmentalization of DC-SIGN+ (red) dermal DCs to lamina propria, Langerin+ (green) Langerhans cells to epithelium, with the complete absence of double-positive cells (yellow, merge). Images were acquired with use of the 20X objective, with a final magnification (optical and digital) of approximately 250X. Shown are the red (panel 1), green (panel 2), and merged (panel 3) channels of the image and the H&E-stained section (panel 4). (B) The vast majority of DC-SIGN+ cells (green) also express mannose receptor (MR) (yellow arrows, panel B3) and are distributed throughout the lamina propria. Images were taken with the use of a 20X objective, with a final magnification of 200X. Shown are green (panel 1), red (panel 2), and merged channels (panel 3) and a merge-DIC overlay (panel 4). Fluorescence images were acquired with a Nikon Eclipse 600 microscope equipped with a color high-resolution CCD camera and PC running Image Pro software. Images were sharpened with the use of 2D-deconvolution software.
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Figure 2. Langerhans cells, dermal DCs, and B-cells contribute to the pool of CD83+ mature DCs in CP. Shown is double-immunofluorescence staining of representative CP gingival tissue. (A) Several immature CD1a+ (green, panel 1) DCs at the junction of epithelium and lamina propria co-express CD83 (yellow arrows, panel 3), suggestive of in situ maturation. Images were taken with the use of a 20X objective, at a final magnification of 200X. (B) Several DC-SIGN+ (green, panel 1) dermal DCs in the lamina propria co-express CD83 (yellow arrows, panel 3). Final magnification (optical and digital), 250X. (C) The vast majority of CD19+ B-cells (green, panels 1, 5, 9) and mature CD83+ DCs (red, panels 2, 6, 10) are single-positive; however, co-expression (yellow arrows, panels 3, 7, 11) is also evident. Final magnification (optical and digital): 200X (panels 1–4); 400x (panels 5–12).
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Figure 3. Mature DCs are associated with large clusters of CD4+ T-cells in chronic periodontitis. Fluorescence images (0.25-µ optical sections) were acquired by confocal laser scanning microscopy, with the use of an epifluorescence microscope (Nikon E800, Japan) integrated to a confocal laser scanning system (BioRad Radiance 2000, Hercules, CA, USA). Shown are the CD4+ T-cells (green, panels A1, B1, C1), CD83+ mature DCs (red, panels A2, B2, C2), and merged channels (yellow, panels A3, C3), indicating co-localization of T-cells and DCs. Also shown is a mature DC alone (panels B2, B3). Final magnification (optical and digital): 400x (A); 1500X (B, C)
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Journal of Dental Research, Vol. 82, No. 9,
736-741 (2003)
DOI: 10.1177/154405910308200915
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