Lipoxygenase products from polymorphonuclear leukocytes and epithelial cells of human saliva

doi:10.1016/0003-9861(87)90572-8

Archives of Biochemistry and Biophysics

Volume 257, Issue 2, September 1987, Pages 321-327

https://doi.org/10.1016/0003-9861(87)90572-8 Get rights and content

Abstract

Physiological mixtures of salivary cells from 10 volunteers were examined for lipoxygenase products of exogenous arachidonic acid, the intracellular penetrability of which was enhanced by ethanol. Leukotriene B₄ and its nonenzymatically produced isomers as well as 5-, 12-, and 15-hydroxyeicosatetraenoic acids (HETE) were produced in all samples. Their identity was confirmed by retention time on reverse-phase high-performance liquid chromatography, ultraviolet spectroscopy, and in the case of 12-HETE, by gas chromatography/mass spectrometry. In order to determine the absolute stereochemistry of 12-HETE, its (—)-menthoxycarbonyl derivative was prepared and subjected to oxidative ozonolysis, methylation, and finally gas chromatography. The results indicate that the original configuration is 12(S)-HETE. The production of leukotriene B₄ correlated directly with the age of the donor while production of 15-HETE varied inversely with age. Alternative stimulation of lipoxygenase production by ionophore A23187 also resulted in substantial amounts of leukotriene B₄ but the amount of 12-HETE produced was less than 2% of the amount generated with the ethanol/arachidonic acid method. This would suggest that 12-HETE production took place predominantly in cells which were not capable of being stimulated by the ionophore. These results point to the epithelial cells, rather than platelets, as the source. The presence of epithelial cells and neutrophils in the saliva, both of which have active lipoxygenase activity, provide the potential for cell interactions of a regulatory nature.

References (40)

E. Lantzman et al.
Oral. Surg
(1970)
F. Fitzpatrick et al.
J. Biol. Chem
(1984)
M.S. Klempner et al.
Blood
(1980)
F.A. Green et al.
Biochem. Biophys. Res. Commun
(1986)
P. Borgeat et al.
J. Biol. Chem
(1979)
M. Hamberg
Anal. Biochem
(1971)
M. Hamberg
Biochem. Biophys. Res. Commun
(1983)
B. Rigas et al.
Biochem. Biophys. Res. Commun
(1983)
J.Y. Vanderhoek et al.
J. Biol. Chem
(1985)
M.I. Siegel et al.
Biochem. Biophys. Res. Commun
(1979)

W.L. Smith et al.

J. Biol. Chem

(1972)

E.J. Goetzl et al.

Prostaglandins

(1980)

P.Y.K. Wong et al.

J. Biol. Chem

(1984)

D.E. Wright

Arch. Oral. Biol

(1964)

C.R. Schiott et al.

J. Periodont. Res

(1970)

D.A. Woolweaver et al.

J. Dent. Res

(1972)

D.R. Miller et al.

J. Clin. Periodontol

(1984)

M.J. Kowolik et al.

J. Periodont. Res

(1980)

P.A. Murray et al.

J. Periodont. Res

(1980)

M.P. Hase et al.

J. Periodont. Res

(1979)

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Citation Excerpt :
We could not detect this metabolite in the ileum, jejunum, colon, gastrocnemius, brain or lung of mice, indicating that 13-HODE-EA is not readily detectable in mice. However, we could find 13-HODE-EA in human saliva samples, which were previously shown to contain 15-LO metabolites [34,35] and NAEs [36]. Indeed, 13-HODE-EA levels were in the mid nM range (Table 3), supporting the hypothesis that this metabolite plays physiological roles.
N-Arachidonoyl-ethanolamine (AEA) is an endocannabinoid (eCB) and endogenous lipid mimicking many of the effects of Δ⁹-tetrahydrocannabinol, notably on brain functions, appetite, pain and inflammation. The eCBs and eCB-like compounds contain fatty acids, the main classes being the monoacylglycerols and the N-acyl-ethanolamines (NAEs). Thus, each long chain fatty acid likely exists under the form of a monoacylglycerol and NAE, as it is the case for arachidonic acid (AA) and linoleic acid (LA). Following their biosynthesis, AA and AEA can be further metabolized into additional eicosanoids, notably by the 15-lipoxygenase pathway. Thus, we postulated that NAEs possessing a 1Z,4Z-pentadiene motif, near their omega end, would be transformed into their 15-lipoxygenase metabolites. As a proof of concept, we investigated N-linoleoyl-ethanolamine (LAE). We successfully synthesized LEA and LEA-d₄ as well as their 15-lipoxygenase-derived derivatives, namely 13-hydroxy-9Z,11E-octadecadienoyl-N-ethanolamine (13-HODE-EA) and 13-HODE-EA-d₄, using Novozyme 435 immobilized on acrylic resin and soybean lipoxygenase respectively. We also show that both human 15-lipoxygenase-1 and -2 can biosynthesize 13-HODE-EA. Co-incubation of LEA and LA with either human 15-lipoxygenase led to the biosynthesis of 13-HODE-EA and 13-HODE in a ratio equal to or greater than 3:1, indicating that LEA is preferred to LA by these enzymes. Finally, we show that 13-HODE-EA is found in human saliva and skin and is a weak although selective TRPV1 agonist. The full biological importance of 13-HODE-EA remains to be explored.
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Sexual orientation is encoded within immune-cell subsets (ICS) of mucosal and epithelial tissues. Gender orientation may be encoded within other ICS. Many immune cells: recognize and react to H-Y and H-X antigens; and enact these perceptions and reactions in accord with the perceiver's and the perceived's MHC haplotype, XX or XY status, and immune-self recognition. Non-heterosexual orientations derive from excessive cross-priming, accompanied by clonal deletions, clonal expansions, anergy and tolerance. For at least some tissues, cross-priming sufficient to induce altered orientations may occur during critical periods of immunological development and can occur during fetal and infant development via maternal–fetal transfusion, placental pathology, and impaired maternal nutrient-status or via excessive peripheral apoptosis during postnatal illness. Mast cell interactions with neurons illustrate how mucosal perceptions can be transduced into neuronal signals that modulate CNS events. This hypothesis is testable by mixed-lymphocyte reactions in appropriate cell subsets. Dendritic-cell immunizations are a potential therapy.
Lipoxygenase products in human saliva: Patients with oral cancer compared to controls
1995, Free Radical Biology and Medicine
Lipoxygenase products were quantified in human mixed saliva and in saliva fractions obtained from a parotid or submandibular gland using gas chromatography-mass spectrometry and stable isotope dilution. In glandular saliva, only linoleic acid was detected at levels of 20–30 ng/ml. In contrast, mixed saliva showed a linoleic acid concentration of around 300 ng/ ml, arachidonic acid levels of around 30 ng/ml, hydroxyoctadecadienoic acid (HODE) levels between 5 and 10 ng/ml, and hydroxyeicosatetraenoic acid (HETE) levels up to 25 ng/ml. By far the most abundant HETE was 12-HETE, and incubation experiments with arachidonic acid showed the presence of a substantial 12-lipoxygenase activity in human mixed saliva, but not in saliva fractions. This activity was identified as 12(S)-lipoxygenase activity by chiral analysis of the reaction product. Investigating mixed saliva and glandular saliva of patients with squamous cell carcinoma in the upper aerodigestive tract and of controls, most patients showed elevated levels of free arachidonic acid and elevated HETE levels. Besides a moderate increase in 12-HETE levels, markedly elevated concentrations of 5-HETE and 15-HETE were observed for the carcinoma patients. The level of free arachidonic acid and the quantitative HETE profile appear to be good markers for the inflammatory processes occurring in the oral mucosa and in saliva in response to the development of squamous cell carcinoma.
Generation and metabolism of lipoxygenase products in normal and membrane-damaged cultured human keratinocytes
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The production and metabolism of lipoxygenase eicosanoids were studied in cultured human keratinocytes. The identity of these eicosanoid structures was established by a variety of chromatographic and analytical techniques. Normal cultured keratinocytes did not produce lipoxygenase eicosanoids either spontaneously or when given arachidonic acid in the presence of permeabilizing concentrations of ethanol or dimethyl sulfoxide. Freeze-thawing of human neonatal and adult keratinocytes resulted in a rapid release of linoleic and arachidonic acids over time. Activation of a latent 15-lipoxygenase was demonstrated by the synthesis of 15-hydroryeicosatetraenoic acid (15-HETE) and 13-hydroxyoctadecadienoic acid, and both these products were greatly increased in amount when the corresponding fatty acid precursor was added. Eicosanoid production by cells of newborn and adult origin was indistinguishable. Rapid metabolism of exogenous 15-HETE by normal keratinocytes was observed. Measurable quantities of esterified 15-HETE were found after 1 min, but by 18–20 h all the esterified 15-HETE was degraded to the extent that 80% of the recovered radioactivity was found in water-soluble form. In contrast, when labeled or unlabeled 5-HETE was used a much larger fraction was esterified intact (30% as opposed to 10%) and at the end of 18–20 hours a substantial peak of esterified 5-HETE remained. Intact esterified [³H] HETE were recovered only in the triacylglycerol fraction. The key findings that ω-6 lipoxygenase products are generated but not esterified by membrane-damaged keratinocytes, whereas these products are esterified but not generated by normal keratinocytes, may be of importance in transcellular metabolic control.
Lipoxygenase activities of the epithelial cells of the human buccal cavity
1989, Biochemical and Biophysical Research Communications
The lipoxygenase activities of cultured human buccal epithelial cells and cells taken ex vivo from the human buccal cavity were compared. Lipoxygenation by cultured cells exhibited exclusively ω-6 positional specificity. A membrane-damaging event such as freezing was required for activation. In contrast, after simple addition of arachidonic acid, the ex vivo buccal cells produced predominantly 12-hydroxyeicosatetraenoic acid. Chromatography on chiral columns yielded products which had an (S) configuration at the site of oxygen insertion. 5-hydroxyeicosatetraenoic acid was transformed by ex vivo buccal cells to 5(S), 12(S)-dihydroxyeicosatetraenoic acid. These products could have a role in modulation of inflammatory states of the oral cavity.
Increased levels of cysteinyl-leukotrienes in saliva, induced sputum, urine and blood from patients with aspirin-intolerant asthma
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A diagnosis of aspirin-intolerant asthma requires aspirin provocation in specialist clinics. Urinary leukotriene E₄ (LTE₄) is increased in aspirin-intolerant asthma. A study was undertaken to investigate new biomarkers of aspirin intolerance by comparing basal levels of cysteinyl-leukotrienes (CysLTs) and leukotriene B₄ (LTB₄) in saliva, sputum and ex vivo stimulated blood in subjects with aspirin-intolerant and aspirin-tolerant asthma. The effects of aspirin- and allergen-induced bronchoconstriction on leukotriene levels in saliva and ex vivo stimulated blood were also compared with the effects of the provocations on urinary mediators.
Induced sputum, saliva, urine and blood were obtained at baseline from 21 subjects with asthma. At a separate visit, 11 subjects showed a positive response to lysine-aspirin inhalation and 10 were aspirin tolerant. Saliva, blood and urine were also collected on the provocation day. Analyses of CysLTs and LTB₄ and the prostaglandin D₂ metabolite 9α, 11β-prostaglandin F₂ were performed and the fraction of exhaled nitric oxide was measured.
Subjects with aspirin-intolerant asthma had higher exhaled nitric oxide levels and higher baseline levels of CysLTs in saliva, sputum, blood ex vivo and urine than subjects with aspirin-tolerant asthma. There were no differences in LTB₄ levels between the groups. Levels of urinary LTE₄ and 9α, 11β-prostaglandin F₂ increased after aspirin provocation whereas leukotriene levels in saliva and ex vivo stimulated blood did not increase.
These findings support a global and specific increase in CysLT production in aspirin-intolerant asthma. Measurement of CysLTs in saliva has the potential to be a new and convenient non-invasive biomarker of aspirin-intolerant asthma.

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^☆: Supported by the Swedish Medical Research Council Grant 03X-07135.

²: On leave from the Departments of Medicine and Microbiology, State University of New York at Buffalo and the Veterans Administration Medical Center, Buffalo, New York.

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Lipoxygenase products from polymorphonuclear leukocytes and epithelial cells of human saliva☆

Abstract

Oral. Surg

J. Biol. Chem

Blood

Biochem. Biophys. Res. Commun

J. Biol. Chem

Anal. Biochem

Biochem. Biophys. Res. Commun

Biochem. Biophys. Res. Commun

J. Biol. Chem

Biochem. Biophys. Res. Commun

J. Biol. Chem

Prostaglandins

J. Biol. Chem

Arch. Oral. Biol

J. Periodont. Res

J. Dent. Res

J. Clin. Periodontol

J. Periodont. Res

J. Periodont. Res

J. Periodont. Res