Monocyte-T-Lymphocyte Co-culture model in Periodontitis

Objectives: CD4+ lymphocytes and monocytes (M) play crucial roles in the etiopathogenesis of periodontitis. Recently our group could show spontaneous ex vivo CD4+ lymphocyte migration to be reduced in chronic periodontitis. The aim of this ex vivo co-culture study was to evaluate migration (LM) of lymphocytes taken from chronic periodontitis (cP) patients in monocyte supernatants (MS) challenged by LPS (E. coli) (MS-LPS) compared to healthy controls (H). Methods: After informed consent CD4+ lymphocytes positively and M negatively were selected immunomagnetically from peripheral blood taken from 10 patients with cP (CD4+cP, McP) and 10 healthy controls without cP (CD4+H, MH). M were incubated with 0.3µl LPS /ml (MS-LPS) and without as control (MS) for 12 h in RPMI 1640. In a time-lapse video-recorded 3-D-collagen matrix assay (1 h) 30 CD4+ cells were selected randomly and LM due to MS vs. MS-LPS were evaluated as to the percentage of cells migrating (%) each in 8 crossover trials per patient and control. Results: In the experiments without LPS challenge CD4+ LM in MS was reduced in the cP group in comparison to the H group (CD4+HMSH: 41% vs. CD4+cPMSH: 38,7%; CD4+HMScP: 43,2% vs. CD4cPMScP: 40,3%). LPS challenge led to enhanced LM in CD4+ cells in the cP group compared to the H group (CD4+H-MSH-LPS: 35,4% vs. CD4+cP-MSH-LPS: 40,7%; CD4+H-MScP-LPS: 33,7% vs. CD4+cP-MScP-LPS: 38,6%). Conclusions: Ex vivo Periodontitis CD4+ lymphocyte migration is reduced due to monocyte supernatants from healthy and from Periodontitis subjects. The reaction was vice versa if LPS challenge was given on the monocytes suggesting CD4+ lymphocyte migratory hyperactivity in reaction to MS after LPS challenge within Periodontitis.