Activation of cyclic AMP response element-binding protein (CREB) in aggressive periodontal disease.
OBJECTIVE
To report a novel observation of neutrophil signal transduction abnormalities in patients with localized aggressive periodontitis (LAP) that are associated with an enhanced phosphorylation of the nuclear signal transduction protein cyclic AMP response element-binding factor (CREB).


METHODS
Peripheral venous blood neutrophils of 18 subjects, 9 patients with LAP and 9 race-, sex-, and age-matched healthy controls, were isolated and prepared using the Ficoll-Hypaque density-gradient technique. Neutrophils (5.4 x 10(6)/mL) were stimulated with the chemoattractant FMLP (10(-6) mol/L) for 5 minutes and lysed. Aliquots of these samples were separated by SDS-PAGE (60 microg/lane) on 9.0% (w/v) polyacrylamide slab gels and transferred electrophoretically to polyvinyl difluoride membranes. The cell lysates were immunoblotted with a 1:1,000 dilution of rabbit-phospho-CREB antibody that recognizes only the phosphorylated form of CREB at Ser133. The activated CREB was visualized with a luminol-enhanced chemoluminescence detection system and evaluated by laser densitometry.


RESULTS
In patients with LAP, the average activation of CREB displayed an overexpression for the unstimulated peripheral blood neutrophils of 80.3% (17.5-fold) compared to healthy controls (4.6%).


CONCLUSIONS
LAP neutrophils who express their phenotype appear to be constitutively primed, as evidenced by activated CREB in resting cells compared to normal individuals. The genetically primed neutrophil phenotype may contribute to neutrophil-mediated tissue damage in the pathogenesis of LAP.
Buchmann
Rainer
R
Division of Periodontology, Medical Faculty, University of Düsseldorf, Düsseldorf, Germany. rainer.buchmann@med.uni-duesseldorf.de
Roessler
Ralf
R
Sculean
Anton
A
D010520:Aggressive Periodontitis / Q000097:blood*; D016022:Case-Control Studies; D002630:Chemotactic Factors / Q000494:pharmacology; D017362:Cyclic AMP Response Element-Binding Protein / Q000097:blood*; D003720:Densitometry / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D015151:Immunoblotting; D007834:Lasers; D049408:Luminescent Agents; D008165:Luminol; D008297:Male; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000378:metabolism; D010766:Phosphorylation; D012694:Serine / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D013997:Time Factors
C115163:CREB1 protein, human; D002630:Chemotactic Factors; D017362:Cyclic AMP Response Element-Binding Protein; D049408:Luminescent Agents; D012694:Serine; D009240:N-Formylmethionine Leucyl-Phenylalanine; D008165:Luminol