Home LiteratureArticle Details
PMID: 8388866 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Enhancement of neutrophil-mediated injury to bovine pulmonary endothelial cells by Pasteurella haemolytica leukotoxin.

Infection and immunity ·Vol. 61 ·No. 6 ·1993-06-00 ·Pages 2618-25

Maheswaran SK, Kannan MS, Weiss DJ, Reddy KR, Townsend EL, Yoo HS, Lee BW, Whiteley LO

Abstract

In this study, we used an in vitro coculture system to determine which virulence factor from Pasteurella haemolytica A1 was responsible for augmenting bovine polymorphonuclear neutrophil (PMN)-mediated killing of bovine pulmonary artery endothelial cells (BPAEC). A 51Cr release cytotoxicity assay was used as a measure of BPAEC killing. The mechanisms associated with this BPAEC killing were also studied. Our results demonstrated that the leukotoxin and not the lipopolysaccharide from P. haemolytica was responsible for augmenting the PMN-mediated killing of BPAEC. Furthermore, this augmented killing was related to the stimulation of PMNs by the leukotoxin. Killing of BPAEC by leukotoxin-stimulated PMNs was diminished in the presence of the H2O2 inactivator, catalase. The membrane-permeant H2O2, hydroxyl radical (HO.) scavenger 1,3-dimethyl-2 thiourea, and the HO. scavenger dimethyl sulfoxide but not the myeloperoxidase inhibitor sodium azide attenuated this BPAEC killing. Pretreatment of BPAEC with a 21-aminosteroid (U74500A), a potent iron chelator-antioxidant, provided the most effective protection against BPAEC killing induced by leukotoxin-stimulated PMNs. These data were compatible with the concept that the H2O2 generated by leukotoxin-stimulated PMNs interacts with intracellular iron in the endothelial cell to form highly reactive HO.. We suggest that HO. may be a key factor in BPAEC killing. Furthermore, since the elastase-specific inhibitor N-methoxy-succinyl-Ala-Ala-Pro-Val-chloromethyl ketone (CMK) also attenuated BPAEC killing and both CMK and 1,3-dimethyl-2 thiourea functioned additively in protecting against BPAEC killing, we conclude that both HO. and elastase may jointly contribute to BPAEC killing induced by leukotoxin-stimulated PMNs. This study broadens our understanding of how leukotoxin-stimulated PMNs injure lung endothelial cells and provides new insight into the pathogenesis of bovine pneumonic pasteurellosis.

MeSH Terms
Amino Acid Sequence Animals Bacterial Toxins/pharmacology Cattle Cells, Cultured Endothelium, Vascular/pathology Exotoxins/pharmacology Female Hydrogen Peroxide/pharmacology Hydroxides/pharmacology Hydroxyl Radical Mannheimia haemolytica/pathogenicity Mice Mice, Inbred BALB C Molecular Sequence Data Neutrophils/physiology Tetradecanoylphorbol Acetate/pharmacology
Chemicals
Bacterial Toxins Exotoxins Hydroxides leukotoxin Hydroxyl Radical Hydrogen Peroxide Tetradecanoylphorbol Acetate
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Maheswaran S K
Department of Veterinary PathoBiology, College of Veterinary Medicine, University of Minnesota, St. Paul 55108.
Kannan M S
Weiss D J
Reddy K R
Townsend E L
Yoo H S
Lee B W
Whiteley L O
References (43)
43 references, click to expand
  1. A review of the lesions in shipping fever of cattle.
    Can Vet J. 1981 Jan;22(1):1-8 PMID: 7018660
  2. Volume-controlled bronchopulmonary lavage of normal and pneumonic calves.
    Vet Pathol. 1986 Nov;23(6):673-80 PMID: 3811133
  3. Morphological and morphometrical analysis of the acute response of the bovine alveolar wall to Pasteurella haemolytica A1-derived endotoxin and leucotoxin.
    J Comp Pathol. 1991 Jan;104(1):23-32 PMID: 2019673
  4. Inhibition of polymorphonuclear leukocyte-mediated endothelial cell detachment by antileukoprotease: a comparison with other proteinase inhibitors.
    Immunobiology. 1991 Mar;182(2):117-26 PMID: 1885205
  5. Xanthine oxidase-induced injury to endothelium: role of intracellular iron and hydroxyl radical.
    Am J Physiol. 1989 Nov;257(5 Pt 2):H1640-6 PMID: 2556049
  6. Dimethylthiourea prevents hydrogen peroxide and neutrophil mediated damage to lung endothelial cells in vitro and disappears in the process.
    Free Radic Biol Med. 1989;6(5):457-66 PMID: 2545552
  7. Sequential lesions of experimental bovine pneumonic pasteurellosis.
    Vet Rec. 1985 Oct 26;117(17):438-42 PMID: 3907114
  8. Iron loading of endothelial cells augments oxidant damage.
    J Lab Clin Med. 1990 Oct;116(4):546-54 PMID: 2120376
  9. A review of infectious bovine rhinotracheitis, shipping fever pneumonia and viral-bacterial synergism in respiratory disease of cattle.
    Can J Comp Med. 1982 Jul;46(3):225-63 PMID: 6290011
  10. Mechanisms of neutrophil-mediated killing of endothelial cells.
    J Leukoc Biol. 1990 Jul;48(1):97-102 PMID: 2193079
  11. Attenuation of tumor necrosis factor-induced endothelial cell cytotoxicity and neutrophil chemiluminescence.
    Am Rev Respir Dis. 1990 Nov;142(5):1073-8 PMID: 2173454
  12. Mechanisms of endothelial cell injury.
    J Lab Clin Med. 1991 Nov;118(5):421-6 PMID: 1658167
  13. Effects of Pasteurella haemolytica A1 leukotoxin on bovine neutrophils: degranulation and generation of oxygen-derived free radicals.
    Vet Immunol Immunopathol. 1992 Jun;33(1-2):51-68 PMID: 1321532
  14. Effect of Pasteurella haemolytica-derived endotoxin on pulmonary structure and function in calves.
    Am J Vet Res. 1990 Mar;51(3):433-8 PMID: 2316921
  15. Role of oxygen-derived free radicals and metabolites in leukocyte-dependent inflammatory reactions.
    Am J Pathol. 1982 Jun;107(3):395-418 PMID: 6282132
  16. Bovine pneumonic pasteurellosis: model for Pasteurella haemolytica- and Pasteurella multocida-induced pneumonia in cattle.
    Am J Vet Res. 1984 Dec;45(12):2532-7 PMID: 6395732
  17. Pulmonary response to intratracheal challenge with Pasteurella haemolytica and Pasteurella multocida.
    Can J Comp Med. 1985 Oct;49(4):395-400 PMID: 3907805
  18. Novel 21-amino steroids as potent inhibitors of iron-dependent lipid peroxidation.
    J Biol Chem. 1987 Aug 5;262(22):10438-40 PMID: 3611075
  19. Pasteurella haemolytica lipopolysaccharide-induced arachidonic acid release from and neutrophil adherence to bovine pulmonary artery endothelial cells.
    Am J Vet Res. 1990 Oct;51(10):1635-9 PMID: 2122778
  20. Interactions of cold stress and Pasteurella haemolytica in the pathogenesis of pneumonic pasteurellosis in calves: changes in pulmonary function.
    Am J Vet Res. 1984 Sep;45(9):1764-70 PMID: 6497134
  21. The 21-aminosteroid inhibitors of lipid peroxidation: reactions with lipid peroxyl and phenoxy radicals.
    Free Radic Biol Med. 1989;7(2):125-30 PMID: 2806938
  22. Direct effects of Pasteurella haemolytica lipopolysaccharide on bovine pulmonary endothelial cells in vitro.
    Am J Vet Res. 1989 Sep;50(9):1633-7 PMID: 2802342
  23. Production and partial characterization of monoclonal antibodies to Pasteurella haemolytica A1 capsular polysaccharide and lipopolysaccharide.
    Am J Vet Res. 1988 Nov;49(11):1962-8 PMID: 2470277
  24. Heterogeneity in phagocytic and nitroblue tetrazolium reductive properties of neutrophils from cows.
    Am J Vet Res. 1991 Dec;52(12):2023-8 PMID: 1789516
  25. Adaptation of a colorimetric microtitration assay for quantifying Pasteurella haemolytica A1 leukotoxin and antileukotoxin.
    Am J Vet Res. 1987 Nov;48(11):1559-64 PMID: 3434898
  26. Isolation of granulocytes and mononuclear cells from the blood of dogs, cats, horses and cattle.
    Vet Clin Pathol. 1989;18(2):33-6 PMID: 15156516
  27. Immunohistochemical localization of Pasteurella haemolytica A1-derived endotoxin, leukotoxin, and capsular polysaccharide in experimental bovine Pasteurella pneumonia.
    Vet Pathol. 1990 May;27(3):150-61 PMID: 2191489
  28. Molecular aspects of virulence of Pasteurella haemolytica.
    Can J Vet Res. 1990 Apr;54 Suppl:S48-52 PMID: 1972901
  29. Hydrogen peroxide causes dimethylthiourea consumption while hydroxyl radical causes dimethyl sulfoxide consumption in vitro.
    J Free Radic Biol Med. 1985;1(5-6):415-9 PMID: 3018065
  30. Pentoxifylline-induced modulation of human leukocyte function in vitro.
    Am J Pathol. 1990 Mar;136(3):623-30 PMID: 2316627
  31. Pore-forming cytolysins of gram-negative bacteria.
    Mol Microbiol. 1991 Mar;5(3):521-8 PMID: 2046545
  32. Neutralizing monoclonal antibodies to Pasteurella haemolytica leukotoxin affinity-purify the toxin from crude culture supernatants.
    Microb Pathog. 1991 May;10(5):411-7 PMID: 1753879
  33. Activation of bovine neutrophils by partially purified Pasteurella haemolytica leukotoxin.
    Infect Immun. 1991 Sep;59(9):3126-33 PMID: 1879935
  34. Evidence of species specificity in the cytocidal effects of Pasteurella haemolytica.
    Infect Immun. 1980 Nov;30(2):615-6 PMID: 7439998
  35. Pulmonary endothelial cell killing by human neutrophils. Possible involvement of hydroxyl radical.
    Lab Invest. 1985 Dec;53(6):656-63 PMID: 2999509
  36. Prevention of granulocyte-mediated oxidant lung injury in rats by a hydroxyl radical scavenger, dimethylthiourea.
    J Clin Invest. 1984 Oct;74(4):1456-64 PMID: 6090504
  37. Lung neutrophils in the adult respiratory distress syndrome. Clinical and pathophysiologic significance.
    Am Rev Respir Dis. 1986 Feb;133(2):218-25 PMID: 3004270
  38. Bovine pulmonary endothelial cell damage mediated by Pasteurella haemolytica pathogenic factors.
    Infect Immun. 1990 Jun;58(6):1671-7 PMID: 2341172
  39. 21-Aminosteroid lipid peroxidation inhibitor U74006F protects against cerebral ischemia in gerbils.
    Stroke. 1988 Aug;19(8):997-1002 PMID: 3400111
  40. Alterations in pulmonary morphology and peripheral coagulation profiles caused by intratracheal inoculation of live and ultraviolet light-killed Pasteurella haemolytica A1 in calves.
    Vet Pathol. 1991 Jul;28(4):275-85 PMID: 1949506
  41. Interaction of bovine neutrophils in Pasteurella haemolytica mediated damage to pulmonary endothelial cells.
    Vet Immunol Immunopathol. 1991 Feb;27(4):337-50 PMID: 2038823
  42. Changes in leukocyte populations in pulmonary lavage fluids of calves after inhalation of Pasteurella haemolytica.
    Am J Vet Res. 1985 Dec;46(12):2429-33 PMID: 3909861
  43. Effects of Pasteurella haemolytica lipopolysaccharide on selected functions of bovine leukocytes.
    Am J Vet Res. 1986 Jan;47(1):154-7 PMID: 3946896
Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/iai.61.6.2618-2625.1993
Published
1993-06-00
Pages
2618-25
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC280892
Subset
IM
External Links
PubMed source DOI: 10.1128/iai.61.6.2618-2625.1993 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2