title: A molecular defect in intracellular lipid signaling in human neutrophils in localized aggressive periodontal tissue damage.
abstract: Host defense mechanisms are impaired in patients with congenital neutrophil (polymorphonuclear neutrophils (PMN)) defects. Impaired PMN chemotaxis is observed in localized aggressive periodontitis (LAP), a familial disorder characterized by destruction of the supporting structures of dentition. In the present studies, we sought evidence for molecular events underlying this aberrant human PMN phenotype. To this end, PMN transendothelial migration and superoxide anion generation were assessed with LAP patients and asymptomatic family members, as well as patients with other chronic mucosal inflammation. PMN from LAP patients showed decreased transmigration across vascular endothelial monolayers (18 +/- 12% of control, n = 4) and increased superoxide anion generation (358 +/- 37%, p = 0.003). Gene expression was analyzed using oligonucleotide microarrays and fluorescence-based kinetic PCR. cDNA microarray and kinetic-PCR analysis revealed diminished RNA expression of leukocyte-type diacylglycerol (DAG) kinase alpha in PMN from LAP patients (4.6 +/- 1.7 relative units, n = 6, p = 0.007) compared with asymptomatic individuals (51 +/- 27 relative units, n = 7). DAG kinase activity was monitored by DAG phosphorylation and individual DAG molecular species were quantified using liquid chromatography and tandem mass spectrometry-based lipidomics. DAG kinase activity was also significantly decreased (73 +/- 2%, p = 0.007) and correlated with increased accumulation of 1,2-diacyl-sn-3-glycerol substrates (p = 0.01). These results implicate defects in both PMN transendothelial migration and PMN DAG kinase alpha signaling as disordered functions in LAP. Moreover, they identify a potential molecular lesion in PMN signal transduction that may account for their aberrant responses and tissue destruction in this disease.
authors.affiliation: Department of Anesthesiology, Perioperative and Pain Medicine, Center for Experimental Therapeutics and Reperfusion Injury, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Karsten
authors.initials: K
authors.lastname: Gronert
authors.forename: Alpdogan
authors.initials: A
authors.lastname: Kantarci
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Levy
authors.forename: Clary B
authors.initials: CB
authors.lastname: Clish
authors.forename: Sabine
authors.initials: S
authors.lastname: Odparlik
authors.forename: Hatice
authors.initials: H
authors.lastname: Hasturk
authors.forename: John A
authors.initials: JA
authors.lastname: Badwey
authors.forename: Sean P
authors.initials: SP
authors.lastname: Colgan
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Van Dyke
authors.forename: Charles N
authors.initials: CN
authors.lastname: Serhan
mesh_terms: D000293:Adolescent; D000328:Adult; D002464:Cell Migration Inhibition; D002634:Chemotaxis, Leukocyte / Q000235:genetics; D002908:Chronic Disease; D019852:Diacylglycerol Kinase / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D004075:Diglycerides / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D004730:Endothelium, Vascular / Q000378:metabolism / Q000473:pathology; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D007424:Intracellular Fluid / Q000201:enzymology / Q000378:metabolism* / Q000502:physiology; D007975:Leukotriene B4 / Q000502:physiology; D008297:Male; D009504:Neutrophils / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D020411:Oligonucleotide Array Sequence Analysis; D010518:Periodontitis / Q000201:enzymology / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D015398:Signal Transduction / Q000235:genetics; D015854:Up-Regulation / Q000235:genetics
chemical_list: D004075:Diglycerides; D007975:Leukotriene B4; D019852:Diacylglycerol Kinase