Interleukin-1 and interleukin-1 inhibitor production by human adherent cells stimulated with periodontopathic bacteria

This study examined the effect of the putative periodontopathic bacteria Bacteroides gingivalis and Fusobacterium nucleatum on the production of interleukin-1 (IL-1) and IL-1 inhibitors by human plastic-adherent mononuclear cells from normal donors. Fusobacterium mortiferum was used as a non-oral, non-pathogenic control organism. Unstimulated adherent cells spontaneously secreted an IL-1 inhibitor, whereas stimulation with B. gingivalis induced the synthesis and secretion of IL-1. With both fusobacteria IL-1 was present in the intracellular environment, whereas the predominant secretory product was either IL-1 or an IL-1 inhibitor. These results suggest that bacteria are capable of modulating cytokine production by monocytes and may thereby alter the local immune response.     

Release of cytokines by stimulated peripheral blood mononuclear cells in chronic periodontitis

Objective

The emergence of periodontal medicine increased interest in defining the behaviour of peripheral blood cells in periodontitis subjects in comparison with healthy group. The aim of this study was to evaluate the levels of interleukin (IL)-8, tumour necrosis factor-α (TNF-α), IL-6 and IL-10 released by Escherichia coli lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (PBMC) obtained from the peripheral blood of chronic periodontitis subjects.

Design

PBMC samples were isolated from 19 systemically healthy donors, divided into generalized chronic periodontitis (n = 10) and healthy (n = 9) subjects. Cells were incubated for 24–48 h in 500 μL wells containing RPMI 1640 and stimulated with 1.0 ng/mL of E. coli LPS. Supernatants were used to quantify the amounts of IL-8, TNF-α, IL-6 and IL-10 released using enzyme-linked immunosorbent assay (ELISA).

Results

PBMC cells from periodontitis subjects released higher levels of TNF-α and IL-6 than those from healthy subjects (P < 0.05). Conversely, the supernatants of the stimulated PBMC cells obtained from healthy subjects presented higher amounts of IL-8 than those from periodontitis (P < 0.05). No differences were observed in the levels of IL-10 (P > 0.05) between groups.

Conclusion

In conclusion, the results of the present study showed that E. coli LPS-stimulated PBMC from subjects with periodontitis present a different pattern of cytokine release when compared to PBMC from healthy subjects. This phenomenon could have implications locally, in periodontitis, as well as in systemic diseases.     

CD45RA and CD45RO positive CD4 cells in human peripheral blood and periodontal disease tissue before and after stimulation with periodontopathic bacteria

Flow cytometric analysis was used to examine naive and primed or memory CD4 cells extracted from periodontal lesions compared with cells from peripheral blood of healthy subjects before and after stimulation with the periodontopathic bacteria, Porphyromonas gingivalis and Fusobacterium nucleatum. In peripheral blood, approximately 60% and 40% of CD4 cells were CD45RO+ and CD45RA+ respectively at day 0. Phytohaemagglutinin (PHA) induced CD45RO expression on almost 100% of CD4 cells. However, P. gingivalis and F. nucleatum stimulation did not cause any significant change in percentage of CD45RO+ CD4 cells except for a loss of antigen at day 6 together with re-expression at day 7, which also occurred on cells cultured in medium only. CD45RA expression on PHA and bacterial-stimulated peripheral blood CD4 cells remained fairly stable for the 10-d culture period. Greater than 90% CD4 cells extracted from healthy or marginal gingivitis (H/MG) and adult periodontitis (AP) lesions were CD45RO+ and this was maintained on AP cells throughout the 6-d culture period, except for a small decrease in the percentage of positive cells induced by P. gingivalis at day 3. Approximately 9% CD4 cells from H/MG tissue were CD45RA+, but about 22% AP cells expressed this antigen, and this increased again in P. gingivalis- and F. nucleatum-stimulated cultures after 3 d. Therefore, in peripheral blood P. gingivalis and F. nucleatum do not act as nonspecific T-cell mitogens and, in AP cells, these bacteria induce changes in phenotype, supporting previous data that although they may be polyclonal B-cell activators, they activate antigen specific T-cells.