Agents and Actions

, Volume 28, Issue 3–4, pp 256–263 | Cite as

In vitro effects of leukotriene B4 (LTB4) on canine PMN effector function(s)

  • D. F. Gruber
  • M. M. D'Alesandro
  • T. L. WaldenJr.
Inflammation and Immunomodulation Meeting Report

Abstract

The canine has become an accepted research model for the examination of a number of human clinical conditions. Despite it's status as a research model, little is known regarding the peripheral effects of inflammatory mediator substances. Products of arachidonic acid metabolism (leukotrienes) are reported capable of altering leukocyte functions. Because of the emerging importance of the canine research model and leukotrienes we examined the effects of leukotriene B4 (LTB4) on severalin vitro functions of isolated canine peripheral polymorphonuclear leukocytes (PMN). Changes in forward angle light scatter properties of the cells were used as one measure of PMN activation. Other functional changes examined following LTB4 pretreatment included chemotactic capability, the electrophysiological state of the cell plasma membrane, and the metabolic oxidative response (i. e. H2O2 production). Random cellular movement of PMNs increased by 120% and 72% following preincubation with 10−7 and 10−9M LTB4, respectively. LTB4 between 10−7 and 10−13M did not significantly alter cellular resting membrane potential. Between 10−7 and 10−9M LTB4 elicited significant levels of cellular H2O2 production. Although significant, H2O2 production was <40% that induced by phorbol myristate acetate (PMA). In numerous respects, caninein vitro PMN responses parallel previous reports of human cell function(s) in the presence of inflammatory mediators and may represent an attractive alternative for investigation of PMN dysfunctions.

Abbreviations

PMN

polymorphonuclear leukocyte

PMA

phorbol myristate acetate

FBS

fetal bovine serum

PBS

phosphate buffered saline

HPLC

high performance liquid chromatography

BRM

baseline random migration

HBSS

Hank's balanced salt solution

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. [1]
    J. A. Simmons, J. A. Salmon and S. Moncada,The release of leukotriene B 4 during experimental inflammation. Biochem. Pharmacol.32, 1353–1359 (1983).PubMedGoogle Scholar
  2. [2]
    A. W. Ford-Hutchinson, M. A. Bray, M. Y. Doig, M. E. Shirley and M. J. Smith,Leukotriene B 4,a potent chemokinetic and aggregating substance released from polymorphonuclear leukocytes. Nature286, 264–265 (1980).PubMedGoogle Scholar
  3. [3]
    M. A. Bray, A. W. Ford-Hutchinson and M. J. H. Smith,Leukotriene B 4:biosynthesis and biological activities. InSRS-A and leukotrienes. (Ed. P. J. Piper) pp. 253–270, J. Wiley and Sons Ltd. 1981.Google Scholar
  4. [4]
    C. L. Malmsten, J. Palmblad, A. Uden, O. Radmark, L. Engstedt and B. Samuelsson,Leukotriene B 4 :A highly potent and stereospecific factor stimulating migration of polymorphonuclear leukocytes. Acta. Physiol. Scand.110, 449–451 (1980).PubMedGoogle Scholar
  5. [5]
    G. D. Gray, G. M. Ohlmann, D. R. Morton and R. G. Schaub,Feline polymorphonuclear leukocytes respond chemotactically to leukotriene B 4 and activated serum but not to f-met-leu-phe. Agents and Actions18, 401–406 (1986).PubMedGoogle Scholar
  6. [6]
    J. Palmblad, C. L. Malmsten, A. M. Uden, L. Radmark, L. Engstedt and B. Samuelsson,Leukotriene B 4 is a potent and stereospecific stimulator of neutrophil chemotaxis and adherence. Blood58, 658–551 (1981).PubMedGoogle Scholar
  7. [7]
    S. Dahlen, S. Bjork, P. Hedquist, K. Arfors, S. Hammarstrom, J. Lindgren and B. Samuelsson,Leukotrienes promote plasma leakage and leukocyte adhesion in postcapillary venules: In vivo effects with relevance to the acute inflammatory response. Proc. Nat. Acad. Sci.78, 3887–3891 (1981).PubMedGoogle Scholar
  8. [8]
    J. T. O'Flaherty, H. J. Showell, E. L. Becker and P. A. Ward,Role of arachidonic acid derivatives in neutrophil aggregation: a hypothesis. Prostaglandins17, 915–927 (1979).PubMedGoogle Scholar
  9. [9]
    B. Ringertz, J. Palmblad and J. Lindgren,Stimulus specific neutrophil aggregation: Evaluation of possible mechanisms for the stimulus response apparatus. J. Lab. Clin. Med.106, 132–140 (1985).PubMedGoogle Scholar
  10. [10]
    R. M. J. Palmer and D. A. Yeats,Leukotriene B: A potent chemotactic agent and stimulus for lysosomal enzyme secretion for human neutrophils (PMN). Br. J. Pharmacol.73, 260–261 (1980).Google Scholar
  11. [11]
    S. J. Feinmark, J. A. Lindren, H. E. Claesson, C. Malmsten and B. Samuelsson,Stimulation of human leukocyte degranulation by leukotriene B 4 and its oxidized metabolites.FEBS Lett. 136, 141–144 (1981).PubMedGoogle Scholar
  12. [12]
    T. E. Rollins, B. Ranolari, M. S. Springer, Y. Guindon, R. Zamboni, C. K. Lau and J. Rokach,Synthetic leukotriene B 4 is a potent chemotaxin but a weak secretagogue for human PMN. Prostaglandins25, 281–289 (1983).PubMedGoogle Scholar
  13. [13]
    J. C. Gay, J. K. Beckman, A. R. Brash, J. A. Oates and J. N. Lukens,Enhancement of chemotactic factor-stimulated neutrophil oxidative metabolism by leukotriene B 4. Blood64, 780–785 (1984).PubMedGoogle Scholar
  14. [14]
    G. M. Omann, A. E. Traynor, A. L. Harris and L. A. Sklar,LTB 4 -induced activation signals and responses in neutrophils are short-lived compared to formylpeptide. J. Immunol.138, 2626–2632 (1987).PubMedGoogle Scholar
  15. [15]
    C. N. Serhan, R. Radin, J. E. Smollen, H. Korchak, J. E. Samuelsson and G. Weissmann,Leukotriene B 4 is a complete secretagogue in human neutrophils: a kinetic analysis. Biochem. Biophys. Res. Commun.107, 1006–1012 (1982).PubMedGoogle Scholar
  16. [16]
    L. Harvath, W. Falk and E. J. Leonard,Rapid quantitation of neutrophil chemotaxis: Use of a polyvinylpyrrolidone-free polycarbonate membrane in a multiwell assembly. J. Immunol. Methods37, 39–45 (1980).PubMedGoogle Scholar
  17. [17]
    D. A. Bass, J. W. Parce, L. R. Dechatelet, P. Szejda, M. C. Seeds and M. Thomas,Flow cytometric studies of oxidative product formation by neutrophils: a graded response to membrane stimulation. J. Immunol.130, 1910–1917 (1983).PubMedGoogle Scholar
  18. [18]
    E. J. Goetzl,Mediators of immediate hypersensitivity derived from arachidonic acid. N. Engl. J. Med.303, 822–825 (1980).PubMedGoogle Scholar
  19. [19]
    R. Huey and T. E. Hugli,Characterization of a C5a receptor on human polymorphonuclear leukocytes (PMN). J. Immunol.135, 2063–2068 (1985).PubMedGoogle Scholar
  20. [20]
    D. W. Goldman and E. J. Goetzl,Heterogeneity of human polymorphonuclear leukocyte receptors for leukotriene B 4:Identification of a subset of high affinity receptors that transduce the chemotactic response. J. Exp. Med.159, 1027–1041 (1984).PubMedGoogle Scholar
  21. [21]
    B. Samulesson, S. Hammarstrom, R. C. Murphy and P. Borgeat,Leukotrienes and slow reacting substance of anaphylaxis. Allergy35, 375–381 (1980).PubMedGoogle Scholar
  22. [22]
    P. Borgeat and B. Samuelsson,Metabolism of arachidonic acid in polymorphonuclear leukocytes. Structure analysis of novel hydroxylated compounds. J. Biol. Chem.245, 7865–7869 (1979).Google Scholar
  23. [23]
    L. T. Williams, R. Snyderman, M. C. Pike and R. J. Lefkowitz,Specific receptor sites for chemotactic peptides on human polymorphonuclear leukocytes. Proc. Nat. Acad. Sci.74, 1204–1208 (1977).PubMedGoogle Scholar
  24. [24]
    S. Aswanikumar, B. Corcoran, A. R. Schiffman, R. J. Day, H. J. Freer, H. J. Showell, E. L. Becker and C. P. Pert,Demonstration of a receptor on rabbit neutrophils for chemotactic peptides. Biochem. Biophys. Res. Commun.74, 810–817 (1977).PubMedGoogle Scholar
  25. [25]
    M. A. Bray, F. A. Cunningham, A. W. Ford-Hutchinson and M. J. H. Smith,Leukotriene B 4:and inflammatory mediator in vivo. Prostaglandins22, 213–222 (1981).PubMedGoogle Scholar
  26. [26]
    P. Bhattacherjee, B. Hammond, J. A. Salmon, R. Stepney and K. E. Eakins,Chemotactic response to some arachidonic acid lipoxygenase products in the rabbit eye. Eur. J. Pharmacol.73, 21–28 (1981).PubMedGoogle Scholar
  27. [27]
    M. J. H. Smith, A. W. Ford-Hutchinson and M. A. Bray,Leukotriene B: a potent mediator of inflammation. J. Pharma. Pharmacol.32, 517–518 (1980).Google Scholar
  28. [28]
    R. D. Camp, A. A. Coutts, M. W. Greaves, A. B. Kay and M. J. Walport,Responses of human skin to intradermal injection of leukotrienes C 4,D 4,and B 4. Br. J. Pharmacol.75, 169 P (1982).Google Scholar
  29. [29]
    H. Redl, P. J. Flynn, H. Lamche, A. Schiesser, G. Schlag and D. E. Hammerschmidt,Aggregation, chemotaxis and chemiluminescence of canine granulocytes. Inflammation7, 67–80 (1983).PubMedGoogle Scholar
  30. [30]
    L. A. Gifford, T. Chernov-Rogan, J. P. Harvey, C. H. Koo, D. W. Goldman and E. J. Goetzl,Recognition of human polymorphonuclear receptors for leukotriene B 4 by rabbit antiidiotypic antibodies to a mouse monoclonal antileukotriene B 4. J. Immunol.138, 1184–1189 (1987).PubMedGoogle Scholar
  31. [31]
    D. W. Goldman, L. A. Gifford, T. Marotti, C. H. Koo and E. J. Goetzl,Molecular and cellular properties of human polymorphonuclear leukocyte receptors for leukotriene B 4. Fed. Proc.46, 200–203 (1987).PubMedGoogle Scholar
  32. [32]
    D. E. Feltner, R. H. Smith and W. A. Marasco,Characterization of the plasma membrane bound GTPase from rabbit neutrophils: Evidence of a Ni-like protein coupled to the formyl peptide, C5a, and leukotriene B 4 chemotaxis receptors. J. Immunol.137, 1961–1970 (1986).PubMedGoogle Scholar
  33. [33]
    E. J. Goetzl and W. C. Pickett,Novel structural determinants of the human neutrophil chemotactic activity of leukotriene B. J. Exp. Med. 153, 482–487 (1981).PubMedGoogle Scholar
  34. [34]
    R. A. Lewis, E. J. Goetzl, J. M. Drazen, N. A. Soter, K. F. Austen and E. J. Corey,Functional characterization of synthetic leukotriene B and its stereochemical isomers. J. Exp. Med.154, 1243–1248 (1981).PubMedGoogle Scholar
  35. [35]
    C. V. Wedmore and T. J. Williams,The control of vascular permeability by polymorphonuclear leukocytes in inflammation. Nature284, 646–650 (1980).Google Scholar
  36. [36]
    M. P. Fletcher,Modulation of the heterogeneous membrane potential response of neutrophils to N-formyl-methionly-leucyl-phenylalanine (fmlp) by leukotriene B 4:evidence for cell recruitment. J. Immunol.136, 4213–4219 (1986).PubMedGoogle Scholar
  37. [37]
    P. H. Naccache, T. F. P. Molski, E. L. Becker, P. Borgeat, S. Picard, P. Vallerand and R. I. Shaafi,Specificity of the effect of lipoxygenase metabolities of arachidonic acid on calcium homeostasis in neutrophils. J. Biol. Chem.257, 8608–8611 (1982).PubMedGoogle Scholar

Copyright information

© Birkhäuser Verlag 1989

Authors and Affiliations

  • D. F. Gruber
    • 1
  • M. M. D'Alesandro
    • 2
  • T. L. WaldenJr.
    • 1
  1. 1.Armed Forces Radiobiology Research InstituteBethesdaUSA
  2. 2.Naval Medical Research Institute Environmental Medicine DepartmentBethesdaUSA

Personalised recommendations