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Home / Equine Research Bank / J Exp Med / Leukotriene generation by eosinophils.
The Journal of experimental medicine1982; 155(2); 390-402; doi: 10.1084/jem.155.2.390

Leukotriene generation by eosinophils.

Abstract: Horse eosinophils purified to greater than 98% generated slow reacting substance (SRS) when incubated with the calcium ionophore A23187. On a per cell basis, eosinophils generated four to five times the SRS produced by similarly treated horse neutrophils. Eosinophil SRS production was inhibited by 5,8,11,14-eicosatetraynoic acid and augmented by indomethacin and arachidonic acid, suggesting that it was a product(s) of the lipoxygenase pathway of arachidonic acid metabolism. Compounds with SRS activity were purified by high-pressure liquid chromatography (HPLC) and identified by ultraviolet spectra, spectral shift on treatment with lipoxygenase, incorporation of [14C]arachidonic acid, gas chromatography-mass spectrometry, and comparison of retention times on HPLC to authentic standards. The eosinophil products characterized were 5-(S), 12-(R)-dihydroxy-6-cis-8, 10-trans-14-cis-eicosatetraenoic acid (leukotriene B4) and its 5-(S), 12-(R)-6-trans and 5-(S), 12-(S)-6-trans isomers, 5-(S)-hydroxy-6-(R)-S-glutathionyl-7,9-trans-11, 14-cis-eicosatetraenoic acid (leukotriene C4) and its 11-trans isomer, and 5-(S)-hydroxy-6-(R)-S-cysteinylglycine-7,9-trans-11,14-cis-eicosatetraenoic acid (leukotriene D4).
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Publication Date: 1982-02-01 PubMed ID: 6120203PubMed Central: PMC2186605DOI: 10.1084/jem.155.2.390Google Scholar: Lookup
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  • P.H.S.

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article reports that horse eosinophils can produce leukotrienes, a type of lipid molecule involved in immune responses, in larger quantities than neutrophils. These substances are identified using various scientific methods including high-pressure liquid chromatography and mass spectrometry.

Overview of the Experiment

  • The researchers purified horse eosinophils, a type of white blood cell, to an exceptional degree (greater than 98%).
  • Eosinophils were then incubated with a calcium ionophore known as A23187, resulting in the generation of a slow reacting substance (SRS).
  • It was found that the eosinophils produced four to five times more SRS than similarly treated horse neutrophils, another type of white blood cell.

Understanding the Mechanism

  • The production of SRS by eosinophils was inhibited by 5,8,11,14-eicosatetraynoic acid and increased when they were exposed to indomethacin and arachidonic acid.
  • This suggests the SRS is a product of the lipoxygenase pathway of arachidonic acid metabolism. That is, when arachidonic acid gets oxidized by lipoxygenases, it results in SRS.

Identification and Characterization of the products

  • The products were purified through high-pressure liquid chromatography (HPLC), a common method for separating substances due to their different interaction rates with a stationary phase in a column.
  • They were then identified using ultraviolet spectra, spectral shift on treatment with lipoxygenase, integration of radiolabeled arachidonic acid, and gas chromatography-mass spectrometry. Implying that these techniques were employed to both confirm the identity of the substances and understand their structures.
  • The specific products characterized were leukotriene B4 and its isomers, leukotriene C4 and its isomer, and leukotriene D4. Leukotrienes are biologically active compounds produced by white blood cells that mediate inflammation in the body. Thus, the large production of these leukotrienes by eosinophils confirms their significant role in inflammation and immune response.

Cite This Article

APA
Jörg A, Henderson WR, Murphy RC, Klebanoff SJ. (1982). Leukotriene generation by eosinophils. J Exp Med, 155(2), 390-402. https://doi.org/10.1084/jem.155.2.390

Publication

The Journal of experimental medicine
ISSN: 0022-1007
NlmUniqueID: 2985109R
Country: United States
Language: English
Volume: 155
Issue: 2
Pages: 390-402

Researcher Affiliations

Jörg, A
    Henderson, W R
      Murphy, R C
        Klebanoff, S J

          MeSH Terms

          • Animals
          • Arachidonic Acids / biosynthesis
          • Arachidonic Acids / isolation & purification
          • Autacoids / antagonists & inhibitors
          • Autacoids / biosynthesis
          • Autacoids / pharmacology
          • Calcimycin / pharmacology
          • Chromones / pharmacology
          • Eosinophils / metabolism
          • Guinea Pigs
          • Horses
          • Humans
          • L-Lactate Dehydrogenase / metabolism
          • Leukotriene B4
          • Mice
          • Mice, Inbred Strains
          • Neutrophils / metabolism
          • SRS-A / biosynthesis
          • SRS-A / isolation & purification

          Grant Funding

          • AI07763 / NIAID NIH HHS
          • AI17758 / NIAID NIH HHS
          • HL25785 / NHLBI NIH HHS

          References

          This article includes 32 references
          1. Augstein J, Farmer JB, Lee TB, Sheard P, Tattersall ML. Selective inhibitor of slow reacting substance of anaphylaxis.. Nat New Biol 1973 Oct 17;245(146):215-7.
            pubmed: 4147765doi: 10.1038/newbio245215a0google scholar: lookup
          2. Flower RJ. Drugs which inhibit prostaglandin biosynthesis.. Pharmacol Rev 1974 Mar;26(1):33-67.
            pubmed: 4208101
          3. Horn BR, Robin ED, Theodore J, Van Kessel A. Total eosinophil counts in the management of bronchial asthma.. N Engl J Med 1975 May 29;292(22):1152-5.
            pubmed: 1124105doi: 10.1056/NEJM197505292922204google scholar: lookup
          4. Conroy MC, Orange RP, Lichtenstein LM. Release of slow reacting substance of anaphylaxis (SRS-A) from human leukocytes by the calcium ionophore A23187.. J Immunol 1976 Jun;116(6):1677-81.
            pubmed: 58045
          5. Jakschik BA, Kulczycki A Jr, MacDonald HH, Parker CW. Release of slow reacting substance (SRS) from rat basophilic leukemia (RBL-1) cells.. J Immunol 1977 Aug;119(2):618-22.
            pubmed: 328779
          6. Murphy RC, Hattox SE, Helbig HR. Application and evaluation of limited mass monitoring with a gas chromatograph mass spectrometer computer system.. Biomed Mass Spectrom 1978 Jul;5(7):444-52.
            pubmed: 678615doi: 10.1002/bms.1200050705google scholar: lookup
          7. Jörg A, Portmann P, Fellay G, Dreyer JL, Meyer J. A rapid and simple method for the isolation of pure eosinophilic leukocytes from horse blood.. Experientia 1978 Dec 15;34(12):1654-6.
            pubmed: 729750doi: 10.1007/BF02034734google scholar: lookup
          8. Borgeat P, Samuelsson B. Transformation of arachidonic acid by rabbit polymorphonuclear leukocytes. Formation of a novel dihydroxyeicosatetraenoic acid.. J Biol Chem 1979 Apr 25;254(8):2643-6.
            pubmed: 429307
          9. Henderson WR, Kaliner M. Mast cell granule peroxidase: location, secretion, and SRS-A inactivation.. J Immunol 1979 Apr;122(4):1322-8.
            pubmed: 87430
          10. Yecies LD, Wedner HJ, Johnson SM, Jakschik BA, Parker CW. Slow reacting substance (SRS) from ionophore A23187-stimulated peritoneal mast cells of the normal rat. I. Conditions of generation and initial characterization.. J Immunol 1979 May;122(5):2083-9.
            pubmed: 87453
          11. Borgeat P, Samuelsson B. Metabolism of arachidonic acid in polymorphonuclear leukocytes. Structural analysis of novel hydroxylated compounds.. J Biol Chem 1979 Aug 25;254(16):7865-9.
            pubmed: 468794
          12. Murphy RC, Hammarström S, Samuelsson B. Leukotriene C: a slow-reacting substance from murine mastocytoma cells.. Proc Natl Acad Sci U S A 1979 Sep;76(9):4275-9.
            pubmed: 41240doi: 10.1073/pnas.76.9.4275google scholar: lookup
          13. Hammarström S, Murphy RC, Samuelsson B, Clark DA, Mioskowski C, Corey EJ. Structure of leukotriene C. Identification of the amino acid part.. Biochem Biophys Res Commun 1979 Dec 28;91(4):1266-72.
            pubmed: 43155doi: 10.1016/0006-291x(79)91203-8google scholar: lookup
          14. Orning L, Hammarström S, Samuelsson B. Leukotriene D: a slow reacting substance from rat basophilic leukemia cells.. Proc Natl Acad Sci U S A 1980 Apr;77(4):2014-7.
            pubmed: 6103542doi: 10.1073/pnas.77.4.2014google scholar: lookup
          15. Bach MK, Brashler JR, Hammarström S, Samuelsson B. Identification of leukotriene C-1 as a major component of slow-reacting substance from rat mononuclear cells.. J Immunol 1980 Jul;125(1):115-7.
            pubmed: 6103909
          16. Falkenhein SF, MacDonald H, Huber MM, Koch D, Parker CW. Effect of the 5-hydroperoxide of eicosatetraenoic acid and inhibitors of the lipoxygenase pathway on the formation of slow reacting substance by rat basophilic leukemia cells; direct evidence that slow reacting substance is a product of the lipoxygenase pathway.. J Immunol 1980 Jul;125(1):163-8.
            pubmed: 6103910
          17. Morris HR, Taylor GW, Piper PJ, Samhoun MN, Tippins JR. Slow reacting substances (SRSs): the structure identification of SRSs from rat basophil leukaemia (RBL-1) cells.. Prostaglandins 1980 Feb;19(2):185-201.
            pubmed: 6104346doi: 10.1016/0090-6980(80)90019-2google scholar: lookup
          18. Bach MK, Brashler JR, Hammarström S, Samuelsson B. Identification of a component of rat mononuclear cell SRS as leukotriene D.. Biochem Biophys Res Commun 1980 Apr 29;93(4):1121-6.
            pubmed: 6104957doi: 10.1016/0006-291x(80)90605-1google scholar: lookup
          19. Clark DA, Goto G, Marfat A, Corey EJ, Hammarström S, Samuelsson B. 11-Trans leukotriene C: a naturally occurring slow reacting substance.. Biochem Biophys Res Commun 1980 Jun 30;94(4):1133-9.
            pubmed: 6772184doi: 10.1016/0006-291x(80)90537-9google scholar: lookup
          20. Ford-Hutchinson AW, Bray MA, Doig MV, Shipley ME, Smith MJ. Leukotriene B, a potent chemokinetic and aggregating substance released from polymorphonuclear leukocytes.. Nature 1980 Jul 17;286(5770):264-5.
            pubmed: 6250050doi: 10.1038/286264a0google scholar: lookup
          21. Lewis RA, Austen KF, Drazen JM, Clark DA, Marfat A, Corey EJ. Slow reacting substances of anaphylaxis: identification of leukotrienes C-1 and D from human and rat sources.. Proc Natl Acad Sci U S A 1980 Jun;77(6):3710-4.
            pubmed: 6106193doi: 10.1073/pnas.77.6.3710google scholar: lookup
          22. Rouzer CA, Scott WA, Hamill AL, Cohn ZA. Dynamics of leukotriene C production by macrophages.. J Exp Med 1980 Nov 1;152(5):1236-47.
            pubmed: 6107327doi: 10.1084/jem.152.5.1236google scholar: lookup
          23. Drazen JM, Austen KF, Lewis RA, Clark DA, Goto G, Marfat A, Corey EJ. Comparative airway and vascular activities of leukotrienes C-1 and D in vivo and in vitro.. Proc Natl Acad Sci U S A 1980 Jul;77(7):4354-8.
            pubmed: 6933488doi: 10.1073/pnas.77.7.4354google scholar: lookup
          24. Rouzer CA, Scott WA, Cohn ZA, Blackburn P, Manning JM. Mouse peritoneal macrophages release leukotriene C in response to a phagocytic stimulus.. Proc Natl Acad Sci U S A 1980 Aug;77(8):4928-32.
            pubmed: 6933538doi: 10.1073/pnas.77.8.4928google scholar: lookup
          25. Samuelsson B, Hammarström S, Murphy RC, Borgeat P. Leukotrienes and slow reacting substance of anaphylaxis (SRS-A).. Allergy 1980 Jul;35(5):375-81.
            pubmed: 6778241doi: 10.1111/j.1398-9995.1980.tb01782.xgoogle scholar: lookup
          26. Sok DE, Pai JK, Atrache V, Sih CJ. Characterization of slow reacting substances (SRSs) of rat basophilic leukemia (RBL-1) cells: effect of cysteine on SRS profile.. Proc Natl Acad Sci U S A 1980 Nov;77(11):6481-5.
            pubmed: 6109281doi: 10.1073/pnas.77.11.6481google scholar: lookup
          27. Sirois P, Borgeat P, Jeanson A, Roy S, Girard G. The action of leukotriene B4 (LTB4) on the lung.. Prostaglandins Med 1980 Dec;5(6):429-44.
            pubmed: 6258182doi: 10.1016/0161-4630(80)90067-1google scholar: lookup
          28. Parker CW, Falkenhein SF, Huber MM. Sequential conversion of the glutathionyl side chain of slow reacting substance (SRS) to cysteinyl-glycine and cysteine in rat basophilic leukemia cells stimulated with A-23187.. Prostaglandins 1980 Nov;20(5):863-86.
            pubmed: 6110221doi: 10.1016/0090-6980(80)90139-2google scholar: lookup
          29. Sirois P, Borgeat P. From slow reacting substance of anaphylaxis (SRS-A) to leukotriene D4 (LTD4).. Int J Immunopharmacol 1980;2(4):281-93.
            pubmed: 6782030doi: 10.1016/0192-0561(80)90028-4google scholar: lookup
          30. Peck MJ, Piper PJ, Williams TJ. The effect of leukotrienes C4 and D4 on the microvasculature of guinea-pig skin.. Prostaglandins 1981 Feb;21(2):315-21.
            pubmed: 6894335doi: 10.1016/0090-6980(81)90149-0google scholar: lookup
          31. Goetzl EJ, Pickett WC. Novel structural determinants of the human neutrophil chemotactic activity of leukotriene B.. J Exp Med 1981 Feb 1;153(2):482-7.
            pubmed: 6264017doi: 10.1084/jem.153.2.482google scholar: lookup
          32. Palmer MR, Mathews WR, Hoffer BJ, Murphy RC. Electrophysiological response of cerebellar Purkinje neurons to leukotriene D4 and B4.. J Pharmacol Exp Ther 1981 Oct;219(1):91-6.
            pubmed: 6270301

          Citations

          This article has been cited 31 times.
          1. Peebles RS Jr. Urine: A Lens for Asthma Pathogenesis and Treatment?. Am J Respir Crit Care Med 2021 Jan 1;203(1):1-3.
            doi: 10.1164/rccm.202007-2899EDpubmed: 32791002google scholar: lookup
          2. Kuranari Y, Tamura R, Mikami S, Ohara K, Toda M, Yoshida K. Severe headache in a patient with meningioma showing extensive dural tail correlates with IgG4-positive plasma cells and eosinophils: A case report and review of literature.. Surg Neurol Int 2018;9:202.
            doi: 10.4103/sni.sni_231_18pubmed: 30386672google scholar: lookup
          3. Angı S, Eken H, Kılıc E, Karaköse O, Balci G, Somuncu E. Effects of Montelukast in an Experimental Model of Acute Pancreatitis.. Med Sci Monit 2016 Aug 1;22:2714-9.
            doi: 10.12659/msm.896919pubmed: 27479458google scholar: lookup
          4. Tian W, Jiang X, Sung YK, Qian J, Yuan K, Nicolls MR. Leukotrienes in pulmonary arterial hypertension.. Immunol Res 2014 May;58(2-3):387-93.
            doi: 10.1007/s12026-014-8492-5pubmed: 24570092google scholar: lookup
          5. Park CY, Kim SW. Idiopathic hypereosinophilic syndrome involving thoracic spine.. J Korean Neurosurg Soc 2010 May;47(5):389-91.
            doi: 10.3340/jkns.2010.47.5.389pubmed: 20539801google scholar: lookup
          6. Maghni K, Nantel F, Lanoue C, Cloutier S, Cristol JP, Cadieux A, Sirois P. Role of airway eosinophilia and eosinophil activation in Sephadex-induced inflammation.. Inflammation 2004 Aug;28(4):195-206.
            doi: 10.1023/b:ifla.0000049044.48102.fcpubmed: 15673161google scholar: lookup
          7. Conroy DM, Sirois P. Early bronchial hyperresponsiveness following injection of sephadex beads in the guinea pig: involvement of platelet activating factor and thromboxane A2.. Inflammation 1999 Oct;23(5):437-48.
            doi: 10.1023/a:1021913009742pubmed: 10466580google scholar: lookup
          8. Chanarin N, Johnston SL. Leukotrienes as a target in asthma therapy.. Drugs 1994 Jan;47(1):12-24.
            doi: 10.2165/00003495-199447010-00002pubmed: 7510618google scholar: lookup
          9. . Cells and inflammation: modern trends and technical outlook.. Klin Wochenschr 1984 May 15;62(10):479-503.
            pubmed: 6748561
          10. Grabbe J, Czarnetzki BM, Mardin M. Release of lipoxygenase products of arachidonic acid from freshly isolated human keratinocytes.. Arch Dermatol Res 1984;276(2):128-30.
            doi: 10.1007/BF00511071pubmed: 6426409google scholar: lookup
          11. Cramer EB, Pologe L, Pawlowski NA, Cohn ZA, Scott WA. Leukotriene C promotes prostacyclin synthesis by human endothelial cells.. Proc Natl Acad Sci U S A 1983 Jul;80(13):4109-13.
            doi: 10.1073/pnas.80.13.4109pubmed: 6408642google scholar: lookup
          12. Henderson WR, Chi EY, Jörg A, Klebanoff SJ. Horse eosinophil degranulation induced by the ionophore A23187. Ultrastructure and role of phospholipase A2.. Am J Pathol 1983 Jun;111(3):341-9.
            pubmed: 6407328
          13. Egwang TG, Befus AD. The role of complement in the induction and regulation of immune responses.. Immunology 1984 Feb;51(2):207-24.
            pubmed: 6363279
          14. Weller PF, Lee CW, Foster DW, Corey EJ, Austen KF, Lewis RA. Generation and metabolism of 5-lipoxygenase pathway leukotrienes by human eosinophils: predominant production of leukotriene C4.. Proc Natl Acad Sci U S A 1983 Dec;80(24):7626-30.
            doi: 10.1073/pnas.80.24.7626pubmed: 6324182google scholar: lookup
          15. Henderson WR, Harley JB, Fauci AS. Arachidonic acid metabolism in normal and hypereosinophilic syndrome human eosinophils: generation of leukotrienes B4, C4, D4 and 15-lipoxygenase products.. Immunology 1984 Apr;51(4):679-86.
            pubmed: 6323307
          16. Lewis RA, Mencia-Huerta JM, Soberman RJ, Hoover D, Marfat A, Corey EJ, Austen KF. Radioimmunoassay for leukotriene B4.. Proc Natl Acad Sci U S A 1982 Dec;79(24):7904-8.
            doi: 10.1073/pnas.79.24.7904pubmed: 6296855google scholar: lookup
          17. Shaw RJ, Cromwell O, Kay AB. Preferential generation of leukotriene C4 by human eosinophils.. Clin Exp Immunol 1984 Jun;56(3):716-22.
            pubmed: 6086189
          18. Potter KA, Leid RW, Kolattukudy PE, Espelie KE. Stimulation of equine eosinophil migration by hydroxyacid metabolites of arachidonic acid.. Am J Pathol 1985 Nov;121(2):361-8.
            pubmed: 4061569
          19. Neill MA, Henderson WR, Klebanoff SJ. Oxidative degradation of leukotriene C4 by human monocytes and monocyte-derived macrophages.. J Exp Med 1985 Nov 1;162(5):1634-44.
            doi: 10.1084/jem.162.5.1634pubmed: 3932580google scholar: lookup
          20. Pincus SH, Cammarata P. Eosinophil adherence to infective larvae of Trichinella spiralis: quantification and modulation.. Immunology 1985 Oct;56(2):219-25.
            pubmed: 3932192
          21. Kay AB. Eosinophils as effector cells in immunity and hypersensitivity disorders.. Clin Exp Immunol 1985 Oct;62(1):1-12.
            pubmed: 3905092
          22. Anthes JC, Bryant RW, Musch MW, Ng K, Siegel MI. Calcium ionophore and chemotactic peptide stimulation of peptidoleukotriene synthesis in DMSO-differentiated HL60 cells.. Inflammation 1986 Jun;10(2):145-56.
            doi: 10.1007/BF00915996pubmed: 3011668google scholar: lookup
          23. Raulf M, Stüning M, König W. Metabolism of leukotrienes by L-gamma-glutamyl-transpeptidase and dipeptidase from human polymorphonuclear granulocytes.. Immunology 1985 May;55(1):135-47.
            pubmed: 2860060
          24. Spada CS, Woodward DF, Hawley SB, Nieves AL, Williams LS, Feldmann BJ. Synergistic effects of LTB4 and LTD4 on leukocyte emigration into the guinea pig conjunctiva.. Am J Pathol 1988 Feb;130(2):354-68.
            pubmed: 2829632
          25. Popper H, Knipping G, Czarnetzki BM, Steiner R, Helleis G, Auer H. Activation and release of enzymes and major basic protein from guinea pig eosinophil granulocytes induced by different inflammatory stimuli and other substances. A histochemical, biochemical, and electron microscopic study.. Inflammation 1989 Apr;13(2):147-62.
            doi: 10.1007/BF00924786pubmed: 2759682google scholar: lookup
          26. Church MK, Benyon RC, Lowman MA, Hutson PA, Holgate ST. Allergy or inflammation? From neuropeptide stimulation of human skin mast cells to studies on the mechanism of the late asthmatic response.. Agents Actions 1989 Jan;26(1-2):22-30.
            doi: 10.1007/BF02126554pubmed: 2653005google scholar: lookup
          27. Guhlmann A, Keppler A, Kästner S, Krieter H, Brückner UB, Messmer K, Keppler D. Prevention of endogenous leukotriene production during anaphylaxis in the guinea pig by an inhibitor of leukotriene biosynthesis (MK-886) but not by dexamethasone.. J Exp Med 1989 Dec 1;170(6):1905-18.
            doi: 10.1084/jem.170.6.1905pubmed: 2584929google scholar: lookup
          28. McColl SR, Krump E, Naccache PH, Caon AC, Borgeat P. Activation of the human neutrophil 5-lipoxygenase by exogenous arachidonic acid: involvement of pertussis toxin-sensitive guanine nucleotide-binding proteins.. Br J Pharmacol 1989 Aug;97(4):1265-73.
            doi: 10.1111/j.1476-5381.1989.tb12588.xpubmed: 2507084google scholar: lookup
          29. Freiburghaus J, Jörg A. Isolation of bovine eosinophils and characterization of their leukotriene formation.. Agents Actions 1990 Aug;31(1-2):16-22.
            doi: 10.1007/BF02003216pubmed: 2178318google scholar: lookup
          30. McEwen BJ, Wilcock BP, Eyre P. The effect of leukotriene B4, leukotriene C4, zymosan activated serum, histamine, tabanid extract and N-formyl-methionyl-leucyl-phenylalanine on the in vitro migration of equine eosinophils.. Can J Vet Res 1990 Oct;54(4):400-4.
            pubmed: 2174292
          31. McEwen BJ. Eosinophils: a review.. Vet Res Commun 1992;16(1):11-44.
            doi: 10.1007/BF01839203pubmed: 1598753google scholar: lookup
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