Journal of Infection and Chemotherapy

, Volume 3, Issue 2, pp 85–89 | Cite as

Antibacterial effects of lactoferrin and a pepsin-generated lactofferrin peptide againstHelicobacter pylori in vitro

  • Natsuko Yamazaki
  • Koji Yamauchi
  • Kouzou Kawase
  • Hirotoshi Hayasawa
  • Kazuyuki Nakao
  • Ichiro Imoto
Original articles

Abstract

Helicobacter pylori is an etiologic agent of gastritis and peptic ulcer disease in humans. We investigated the antibacterial activities of lactoferrin and Lactoferricin® (an antimicrobial peptide derived from lactoferrin) against this bacterium. The minimum bactericidal concentrations of bovine and human lactoferrins were 1.25 to 2.50 mg/mL, while it was greater than 5.0 mg/L for bovine Lactoferricin®. Time-kill studies withH. pylori grown in brucella broth demonstrated that the antibacterial effects of the lactoferrins were dose-dependent within the range of 0.8 to 2.0 mg/mL and began when the cells were in the exponential phase of growth. Iron-saturated lactoferrin did not inhibit the growth ofH. pylori. Bovine Lactoferricin® showed only weak activity againstH. pylori in brucella broth, however, a rapid bactericidal effect was observed in 1% Bacto-peptone medium within 1 hour of exposure at concentrations in the range of 0.1 to 1.0 mg/mL. Under these conditions, bovine lactoferrin showed little effect. Moreover, bovine Lactoferricin® inhibited the urease activity ofH. pylori, which is one of the virulence factors of this bacterium. These results indicate thatH. pylori is susceptible to inhibition and inactivation by both lactoferrin and Lactoferricin® in laboratory media. The antibacterial effect of lactoferrin appeared to be dependent on its iron status, the cellular phase of growth, and has a mechanism of action different from that of Lactoferricin®.

Key words

lactoferrin Lactoferricin® Helicobacter pylori antibacterial effects urease activity 

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References

  1. 1.
    George EB, William KG, Won KL, Joan ML, Anthony RD. Relation ofCampylobacter pyloridis to gastritis and peptic ulcer. J Infect Dis 1986; 153: 664–669.Google Scholar
  2. 2.
    Lee A, Fox J, Hazell S. Pathogenicity ofHelicobacter pylori: a perspective. Infect Immun 1993; 61: 1601–1610.PubMedGoogle Scholar
  3. 3.
    Kato M, Asaka M, Katagiri M, Sukegawa M, Kagaya H, Nishikawa K, et al. Looking for Japanese regimens for the eradication ofHelicobacter pylori. Endoscopia Digestiva 1996; 8: 661–667 (in Japanese).Google Scholar
  4. 4.
    Diker KS, Hascelik G. The bactericidal activity of tea againstHelicobacter pylori. Lett Appl Microbiol 1994; 19: 299–300.Google Scholar
  5. 5.
    Khulusi S, Ahmed HA, Mendall MA, Northfield TC. The effects of unsaturated fatty acid onHelicobacter pylori in vitro. J Med Microbiol 1995; 42: 276–282.PubMedGoogle Scholar
  6. 6.
    Masson PL, Heremans JF, Dive C. An iron-binding protein common to many external secretions. Clin Chim Acta 1966; 13: 735–739.CrossRefGoogle Scholar
  7. 7.
    Brock JH. Lactoferrin in human milk: its role in iron absorption and protection against enteric infection in new born infants. Arch Dis Child 1980; 55: 417–421.PubMedGoogle Scholar
  8. 8.
    Sánchez L, Miguel C, Brock JH. Biological role of lactoferrin. Arch Dis Child 1992; 67: 657–661.PubMedCrossRefGoogle Scholar
  9. 9.
    Bullen JJ, Rogers HJ, Griffiths E. Role of iron in bacterial infection. Curr Top Microbiol Immunol 1987; 80: 1–35.Google Scholar
  10. 10.
    Tomita M, Bellamy W, Takase M, Yamauchi K, Wakabayashi H, Kawase K. Potent antibacterial peptides generated by pepsin digestion of bovine lactoferrin. J Dairy Sci 1991; 74: 4137–4142.PubMedGoogle Scholar
  11. 11.
    Bellamy W, Takase M, Wakabayashi H, Kawase K, Tomita M. Antibacterial spectrum of lactoferricin B, a potent bactericidal peptide derived from the N-terminal region of bovine lactoferrin. J Appl Bacteriol 1992; 74: 432–479.Google Scholar
  12. 12.
    Bellamy W, Takase M, Yamauchi K, Wakabayashi H, Kawase K, Tomita M. Identification of the bactericidal domain of lactoferrin. Biochim Biophys Acta 1992; 1121: 130–136.PubMedGoogle Scholar
  13. 13.
    Ellison RT, Giehl TJ, Laforce FM. Damage of the outer membrane of enteric Gram-negative bacteria by lactoferrin and transferrin. Infect Immun 1988; 56: 2774–2781.PubMedGoogle Scholar
  14. 14.
    Bellamy W, Wakabayashi H, Takase M, Kawase K, Shimamura S, Tomita M. Role of cell-binding in the antibacterial mechanism of lactoferricin B. J Appl Bacteriol 1993; 75: 478–484.PubMedGoogle Scholar
  15. 15.
    Yamauchi K, Tomita M, Giehl TJ, Ellison RT. Antibacterial activity of lactoferrin and a pepsin-derived lactoferrin peptide fragment. Infect Immun 1993; 61: 719–728.PubMedGoogle Scholar
  16. 16.
    Ellison RT, Giehl TJ. Killing of Gram-negative bacteria by lactoferrin and lysozyme. J Clin Invest 1991; 88; 1080–1091.PubMedCrossRefGoogle Scholar
  17. 17.
    Law BA, Reiter B. The isolation bacteriostatic properties of lactoferrin from bovine milk whey. J Dairy Res 1977; 44: 595–561.PubMedCrossRefGoogle Scholar
  18. 18.
    Suzuki T, Yamauchi K, Kawase K, Tomita M, Kiyosawa I, Okonogi S. Collaborative bacteriostatic activity of bovine lactoferrin with lysozyme againstEscherichia coli O111. Agric Biol Chem 1989; 53: 1705–1706.Google Scholar
  19. 19.
    Olivieri R, Bugnoli M, Almellini D, Bianciardi S, Rappuoli R, Bayeli PF et al. Growth ofHelicobacter pylori in media containing cyclodextrins. J Clin Microbiol 1993; 31: 160–162.PubMedGoogle Scholar
  20. 20.
    Takebe S, Kobashi K. Acid urease fromLactobacillus of rat intestine. Chem Pharm Bull 1988; 36: 693–699.PubMedGoogle Scholar
  21. 21.
    Miehlke S, Reddy R, Osato MS, Ward PP, Conneely OM, Graham DY. Direct activity of recombinant human lactoferrin againstHelicobacter pylori. J Clin Microbiol 1996; 34: 2593–2594.PubMedGoogle Scholar
  22. 22.
    Nagasawa T, Kiyosawa I, Kuwahara K. Amount of lactoferrin in human colostrum and milk. J Dairy Sci 1972; 55: 1651–1659.PubMedCrossRefGoogle Scholar
  23. 23.
    Hennart PF, Brasseur DJ, Delogne-Desnoeck JB, Dramaix MM, Robyn CE. Lysozyme, lactoferrin, and secretory immunoglobulin A content in breast milk: influence of duration of lactation, nutrition status, prolactin status, and parity of mother. Am J Nutr 1991; 53: 32–39.Google Scholar
  24. 24.
    Masson PL, Heremans JF. Lactoferrin in milk from different species. Comp Biochem Physiol 1971; 39: 119.Google Scholar
  25. 25.
    Sánchez L, Aranda P, Pérez MD, Calvo M. Concentration of lactoferrin and transferrin throughout lactation in cow's colostrum and milk. Biol Chem Hoppe Seyler 1988; 369: 1005–1008.PubMedGoogle Scholar
  26. 26.
    Anderson BF, Baker HM, Norris GE, Rumball SV, Baker EN. Apolactoferrin structure demonstrates ligand-induced conformational change in transferrins. Nature 1990; 344: 784–787.PubMedCrossRefGoogle Scholar
  27. 27.
    Baker EN, Anderson BF, Baker HM, Haridas M, Jameson GB, Norris GE et al. Structure, function and flexibility of human lactoferrin. Int J Biol Macromol 1991; 13: 122–129.PubMedCrossRefGoogle Scholar
  28. 28.
    Arnold RR, Russell JE, Champion WJ, Gauthier JJ. Bactericidal activity of human lactoferrin: influence of physical condition and metabolic state of the target microorganism. Infect Immun 1981; 32: 655–660.PubMedGoogle Scholar
  29. 29.
    Husson MO, Legrand D, Spik G, Leclerc H. Iron acquisition byHelicobacter pylori: importance of human lactoferrin. Infect Immun 1993; 61: 2694–2697.PubMedGoogle Scholar
  30. 30.
    Worst DJ, Otto BR, Graaff J. Iron-repressible outer membrane proteins ofHelicobacter pylori involved in heme uptake. Infect Immun 1995; 63: 461–465.Google Scholar
  31. 31.
    Catrenich CE, Makin KM. Characterization of the morphologic conversion ofHelicobacter pylori from bacillary to coccoid forms. Scand J Gastroenterol 1991; 26(suppl 181): 58–64.Google Scholar
  32. 32.
    Berry V, Jennings K, Woodnutt G. Bactericidal and morphological effects of amoxicillin onHelicobacter pylori. Antimicrob Agents Chemother 1995; 39: 1859–1861.PubMedGoogle Scholar
  33. 33.
    Tomita M, Takase M, Bellamy W, Shimamura S. A review: the active peptide of lactoferrin. Acta Paediatr Jpn 1994; 36: 585–591.PubMedGoogle Scholar

Copyright information

© JSC/CLJ 1997

Authors and Affiliations

  • Natsuko Yamazaki
    • 1
  • Koji Yamauchi
    • 1
  • Kouzou Kawase
    • 1
  • Hirotoshi Hayasawa
    • 1
  • Kazuyuki Nakao
    • 2
  • Ichiro Imoto
    • 2
  1. 1.Nutritional Science LaboratoryMorinaga Milk Industry, Co., Ltd.ZamaJapan
  2. 2.Third Department of Internal MedicineMie University School of MedicineMieJapan

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