Nitric oxide synthase activity in neutrophils from patients with localized aggressive periodontitis.
BACKGROUND
Localized aggressive periodontitis (LAgP) is associated with neutrophil dysfunction including defective chemotaxis and reduced calcium influx factor activity. Nitric oxide (NO) and its enzyme, nitric oxide synthase (NOS), have been suggested to be involved in chemotaxis. Some reports, however, were unable to detect either NO or NOS in human neutrophils. In this study, we focused on NOS activity in LAgP neutrophils and examined the involvement of NOS in chemotaxis of normal neutrophils and NOS activity in neutrophils from normal subjects and patients with LAgP.


METHODS
Neutrophils from 10 normal subjects and 10 LAgP patients were isolated from peripheral venous blood. Membrane associated-NOS (MA-NOS) and soluble NOS (S-NOS) were extracted from cells with or without FMLP stimulation. NOS activity was measured using the radiolabeled L-arginine to L-citrulline conversion assay.


RESULTS
N(omega)-nitro-L-arginine methyl ester (L-NAME), an inhibitor of NOS, significantly inhibited FMLP-induced chemotaxis (P<0.01) and dibutyryl cGMP, an activator of cGMP-dependent protein kinase, significantly attenuated the inhibition by L-NAME (P<0.01). Unstimulated and FMLP-stimulated MA-NOS activity in LAgP neutrophils was statistically significantly higher than that in normal neutrophils (P<0.05). S-NOS activity in LAgP neutrophils was higher than that in normal neutrophils.


CONCLUSIONS
This study suggests that NOS is present in human neutrophils and may be involved in FMLP-induced chemotaxis in normal neutrophils. NOS activity is increased in LAgP and is negatively correlated to chemotaxis response.
Shibata
K
K
Department of Periodontology and Oral Biology, Boston University Goldman School of Dental Medicine, MA 02118, USA.
Warbington
M L
ML
Gordon
B J
BJ
Kurihara
H
H
Van Dyke
T E
TE
D015126:8,11,14-Eicosatrienoic Acid / Q000031:analogs & derivatives* / Q000378:metabolism; D000293:Adolescent; D000328:Adult; D010520:Aggressive Periodontitis / Q000201:enzymology* / Q000276:immunology*; D000704:Analysis of Variance; D002118:Calcium / Q000378:metabolism; D002120:Calcium Channel Agonists / Q000378:metabolism; D016022:Case-Control Studies; D002634:Chemotaxis, Leukocyte; D005260:Female; D006801:Humans; D008297:Male; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009504:Neutrophils / Q000201:enzymology* / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000378:metabolism*
D002120:Calcium Channel Agonists; D009569:Nitric Oxide; C040776:5,6-epoxy-8,11,14-eicosatrienoic acid; D019001:Nitric Oxide Synthase; D015126:8,11,14-Eicosatrienoic Acid; D002118:Calcium; D019331:NG-Nitroarginine Methyl Ester