title: DNA from periodontopathogenic bacteria is immunostimulatory for mouse and human immune cells.
abstract: Although bacterial DNA (bDNA) containing unmethylated CpG motifs stimulates innate immune cells through Toll-like receptor 9 (TLR-9), its precise role in the pathophysiology of diseases is still equivocal. Here we examined the immunostimulatory effects of DNA extracted from periodontopathogenic bacteria. A major role in the etiology of periodontal diseases has been attributed to Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Peptostreptococcus micros. We therefore isolated DNA from these bacteria and stimulated murine macrophages and human gingival fibroblasts (HGF) in vitro. Furthermore, HEK 293 cells transfected with human TLR-9 were also stimulated with these DNA preparations. We observed that DNA from these pathogens stimulates macrophages and gingival fibroblasts to produce tumor necrosis factor alpha and interleukin-6 in a dose-dependent manner. Methylation of the CpG motifs abolished the observed effects. Activation of HEK 293 cells expressing TLR-9 which were responsive to bDNA but not to lipopolysaccharide confirmed that immunostimulation was achieved by bDNA. In addition, the examined bDNA differed in the ability to stimulate murine macrophages, HGF, and TLR-9-transfected cells. DNA from A. actinomycetemcomitans elicited a potent cytokine response, while DNA from P. gingivalis and P. micros showed lower immunostimulatory activity. Taken together, the results strongly suggest that DNA from A. actinomycetemcomitans, P. gingivalis, and P. micros possesses immunostimulatory properties in regard to cytokine secretion by macrophages and fibroblasts. These stimulatory effects are due to unmethylated CpG motifs within bDNA and differ between distinct periodontopathogenic bacteria strains. Hence, immunostimulation by DNA from A. actinomycetemcomitans, P. gingivalis, and P. micros could contribute to the pathogenesis of periodontal diseases.
authors.affiliation: Institute of Medical Microbiology and Hygiene, Philipps University, Marburg, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Nonnenmacher
authors.forename: Alexander
authors.initials: A
authors.lastname: Dalpke
authors.forename: Stefan
authors.initials: S
authors.lastname: Zimmermann
authors.forename: Lavin
authors.initials: L
authors.lastname: Flores-De-Jacoby
authors.forename: Reinier
authors.initials: R
authors.lastname: Mutters
authors.forename: Klaus
authors.initials: K
authors.lastname: Heeg
mesh_terms: D016976:Aggregatibacter actinomycetemcomitans / Q000235:genetics / Q000276:immunology*; D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D018899:CpG Islands / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D029721:Drosophila Proteins; D005347:Fibroblasts / Q000276:immunology; D005881:Gingiva / Q000166:cytology; D006801:Humans; D015850:Interleukin-6 / Q000096:biosynthesis; D008262:Macrophage Activation; D008264:Macrophages / Q000276:immunology; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D008745:Methylation; D051379:Mice; D010462:Peptostreptococcus / Q000235:genetics / Q000276:immunology*; D010510:Periodontal Diseases / Q000382:microbiology; D016966:Porphyromonas gingivalis / Q000235:genetics / Q000276:immunology*; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism; D051217:Toll-Like Receptor 9; D051193:Toll-Like Receptors; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D004269:DNA, Bacterial; D029721:Drosophila Proteins; D015850:Interleukin-6; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; C495371:TLR9 protein, human; D051217:Toll-Like Receptor 9; D051193:Toll-Like Receptors; D014409:Tumor Necrosis Factor-alpha