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Dendritic-NK Cell Interactions in P. gingivalis-specific Responses

Clinical Research Center for Periodontal Diseases, School of Dentistry, VCU, PO Box 980556, Richmond, VA 23298-0556, USA;

View larger version (10K): [in a new window]   Figure 1. P. gingivalis-stimulated dendritic cells, but not macrophages, induced a rapid IFN- response in peripheral blood mononuclear cells (PBL). PBL from a patient with a normal periodontium were stimulated with 1 x 106 P. gingivalis (Pg) and autologous dendritic cells (DC or mDCs) or macrophages (Ms). Supernatant fluids were collected 24 hrs later, and IFN- production was measured. Additional negative controls that did not differ from the background with PBL alone included: dendritic cells plus PBL, macrophages plus PBL, additional PBL corresponding in number with the dendritic cells and macrophages, and dendritic cells or macrophages with Pg but in the absence of PBL (data not shown). The data are representative of 3 experiments of this type; mean ± SE.

View larger version (22K): [in a new window]   Figure 2. Influence of CD56+ cells, dendritic cells, and IL-12 on early IFN- responses. (A) PBL from a subject with a normal periodontium or CD56+-depleted PBL from the same subject at 1 x 106 cells/culture were stimulated with 1 x 106 P. gingivalis (Pg) with autologous dendritic cells (DC). Supernatant fluids were collected at 24 hrs, and IFN- was measured. These data are representative of 3 experiments of this type. The "-CD56+" represents NK-cell-depleted PBL. (B) The CD56+ cells were positively selected and stimulated with 1 x 106 P. gingivalis and autologous dendritic cells. Supernatant fluids were collected at 24 hrs, and IFN- was measured. These data are representative of 3 experiments. (C) PBL from a subject with a normal periodontium plus autologous dendritic cells were treated with 10 µg/mL anti-IL-12 and stimulation with 1 x 106 P. gingivalis. Supernatant fluids were collected at 24 hrs, and IFN- was measured. The data are representative of 3 experiments of this type; mean ± SE.

View larger version (15K): [in a new window]   Figure 3. Influence of dendritic cells and NK cells on IgG2 anti-P. gingivalis production. (A) PBL from a P. gingivalis (Pg)-seropositive subject with chronic periodontitis ± autologous dendritic cells (DC) and/or follicular dendritic cells (FDCs) were stimulated with 1 x 106 P. gingivalis-anti-P. gingivalis immune complex (Pg-ICs). After a six-day inductive period, the cells were washed to remove any free antigen or antibody, new culture media were added, and supernatant fluids were collected at 14 days. The IgG2 anti-P. gingivalis produced from days 6 to 14 was measured with the supernatant fluids. The data are recorded as the mean ± SE, and * indicates p < 0.05. These data are representative of 3 experiments of this type. (B) PBL from a P. gingivalis-seropositive chronic periodontitis subject or CD56+-depleted PBL from the same subject were held constant at 1 x 106 cells per culture ± follicular dendritic cells, and were stimulated with 1 x 106 P. gingivalis-anti-P. gingivalis immune complexes. Supernatant fluids were collected at 14 days as described above, and IgG2 anti-P. gingivalis production was measured. The data are recorded as the mean ± SE, and * indicates p < 0.05. These data are representative of 3 experiments of this type.