Methods: Murine macrophages were isolated. The cells were co-incubated with 50mg of DNA derived from Pg W83, Pg cells MOI 100:1 or no challenge at 6h. Macrophage RNA was isolated and microarray analysis was performed.
Results: Initial microarray analysis demonstrated that both Pg and Pg DNA showed similar numbers of genes regulated that were involved in the inflammatory response: an average of 1559 (Pg) vs. 1451(DNA). In addition, similar numbers of genes were regulated in the IL-8, chemokine, NF-κB and IL-6 pathways under both conditions. Also, TLR9-/- cells had upregulation of IFNAR1 when challenged with DNA than the WT cells and JAK1 showed similar levels of regulation amongst WT and TLR9-/- cells.
Conclusions: These results suggest that DNA is the major virulence factors in the immune response. Due to type I interferon response in the TLR9-/- cells, we believe that DAI or other unknown DNA receptor(s) may be responsible. Finally, the similar regulation of JAK1 in both cell types challenged with DNA shows that a TLR9 independent mechanism may predominate.
Keywords: Cytokine, Gene expression, Immune response, Inflammatory mediators and Periodontal disease
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