Evidence of an association between functional abnormalities and defective diacylglycerol kinase activity in peripheral blood neutrophils from patients with localized juvenile periodontitis.
Peripheral neutrophils from patients with localized juvenile periodontitis (LJP) show functional abnormalities, such as impaired locomotion and enhanced respiratory burst activity. A defect in intracellular signalling mechanism has been proposed to be responsible for some changes, but direct evidence is lacking. In this study we have determined the activity of diacylglycerol (DAG) kinase, an enzyme controlling the DAG/protein kinase C (PKC) pathway, in crude cytosolic and membrane fractions of neutrophils from 5L JP patients and age and gender-matched normal individuals. No difference was observed in the DAG kinase activity in subcellular fractions from unstimulated cells between the 2 groups. When normal neutrophils were stimulated with N formyl-methionyl-leucyl-phenylalanine (FMLP), the enzyme activity was markedly increased in both subcellular fractions. In contrast, neutrophils from 3 of the 5 LJP patients tested completely failed to rise the DAG kinase activity upon chemoattractant stimulation. These data indicate that in some LJP patients the neutrophil DAG kinase may be defective. To examine whether a decrease in DAG kinase activity could account for some neutrophil abnormalities seen in LJP, normal neutrophils were treated with R59022, a DAG kinase inhibitor, that has been shown to reduce DAG kinase activity in human neutrophils. Upon stimulation with FMLP, R59022-treated normal neutrophils showed significantly reduced chemotactic response and enhanced respiratory burst activity, two typical functional abnormalities featured by LJP cells. It is concluded that a defect in DAG kinase may cause, through an abnormal accumulation of the endogenous PKC activator DAG some of the functional changes observed in neutrophils from LJP patients.
Hurttia
H M
HM
Department of Dentistry, University of Turku, Finland.
Pelto
L M
LM
Leino
L
L
D000293:Adolescent; D000328:Adult; D010520:Aggressive Periodontitis / Q000097:blood / Q000201:enzymology* / Q000276:immunology*; D016022:Case-Control Studies; D002634:Chemotaxis, Leukocyte; D019852:Diacylglycerol Kinase; D005260:Female; D006801:Humans; D009240:N-Formylmethionine Leucyl-Phenylalanine; D018375:Neutrophil Activation; D009504:Neutrophils / Q000201:enzymology* / Q000378:metabolism / Q000502:physiology; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000097:blood / Q000378:metabolism*; D016897:Respiratory Burst; D015398:Signal Transduction
D009240:N-Formylmethionine Leucyl-Phenylalanine; D017853:Phosphotransferases (Alcohol Group Acceptor); D019852:Diacylglycerol Kinase