Abstract
Four avian heterophil antimicrobial cationic peptides (Chicken Heterophil Peptides 1 and 2, and Turkey Heterophil Peptides 1 and 3) were evaluated for in vitro microbicidal activity against selected avian pathogens and human pathogens which are harbored by birds. At concentrations of 16-2 μg/ml, all four avian peptides effected a greater than 90% reduction in the survival of Candida albicans, Salmonella enteriditis, and Campylobacter jejuni. None of the peptides, including the known antimicrobial peptide protamine (used as a positive control), were able to reduce the survival of Pasteurella multocida by 90% at the maximum peptide concentration (16 μg/ml) tested. At 16 μ/ml, the turkey peptide THP3 did not effect a 90% reduction in survival of Bordetella avium, Escherichia coli, or Salmonella typhimurium, while all of the other peptides tested were effective at this concentration or less. This peptide, THP3, does not share the same homologous amino acid sequence shared by the other three peptides. Under our experimental conditions, none of the peptides neutralized Infectious Bronchitis Virus, an enveloped coronavirus of chickens.
Keywords: Beta-defensin, Avian heterophils, Chicken, Turkey, Antimicrobial peptides
References
- Ahmad I., Kleven S.H., Avakian A.P., Glisson J.R. Sensitivity and specificity of Mycoplasma gallisepticum agglutination antigens prepared from medium with artificial liposomes substituting for serum. Avian Dis. 1988;32:519–526. [PubMed] [Google Scholar]
- Black R.E., Levine M.M., Clements M.L., Hughes T.P., Blaser M.J. Experimental Campylobacter jejuni infection in humans. J. Infect. Dis. 1988;157:472–479. doi: 10.1093/infdis/157.3.472. [DOI] [PubMed] [Google Scholar]
- Breton-Gorius J., Coquin Y., Guichard J. Cytochemical distinction between azurophils and catalase-containing granules in leukocytes. Lab. Invest. 1978;38:21–31. [PubMed] [Google Scholar]
- Diamond G., Zasloff M., Eck H., Brasseur M., Maloy W.L., Bevins C.L. Tracheal antimicrobial peptide, a cysteine-rich peptide from mammalian tracheal mucosa: peptide isolation and cloning of a cDNA. Biochemistry. 1991;88:3952–3956. doi: 10.1073/pnas.88.9.3952. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Duvick J.P., Rood T., Rao A.G., Marshak D.R. Purification and characterization of a novel antimicrobial peptide from Maize (Zea mays L.) kernels. J. Biol. Chem. 1992;267:18814–18820. [PubMed] [Google Scholar]
- Eisenhauer P.B., Harwig S.S.L., Szklarek D., Ganz T., Selsted M.E., Lehrer R.I. Purification and antimicrobial properties of three defensins from rat neutrophils. Infect. Immun. 1989;57:2021–2027. doi: 10.1128/iai.57.7.2021-2027.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Evans E.W., Beach F.G., Wunderlich J., Harmon B.G. Isolation of antimicrobial peptides from avian heterophils. J. Leukoc. Biol. 1994;56:661–665. doi: 10.1002/jlb.56.5.661. [DOI] [PubMed] [Google Scholar]
- Gelb J., Jr., Perkins B.E., Rosenberger J.K., Allen P.H. Serologic and cross-protection studies with several infectious bronchitis virus isolates from Delmarva-reared broiler chickens. Avian Dis. 1981;25:655–666. [PubMed] [Google Scholar]
- Harmon B.G., Glisson J.R., Nunnally J.C. Turkey macrophage and heterophil bactericidal activity against Pasteurella multocida. Avian Dis. 1992;36:986–991. [PubMed] [Google Scholar]
- Harwig S.S.L., Swiderek K.M., Kokryakov V.N., Tan L., Lee T.D., Panyutich E.A., Aleshina G.M., Shamova O.V., Lehrer R.I. Gallinacins: Cysteine-rich antimicrobial peptides of chicken leukocytes. FEBS Lett. 1994;342:281–285. doi: 10.1016/0014-5793(94)80517-2. [DOI] [PubMed] [Google Scholar]
- Jones D.E., Bevins C.L. Defensin-6 mRNA in human Paneth cells: implications for antimicrobial peptides in host defense of the human bowel. FEBS Lett. 1993;315:187–192. doi: 10.1016/0014-5793(93)81160-2. [DOI] [PubMed] [Google Scholar]
- Kimbrell D.A. Insect antibacterial proteins: not just for insects and against bacteria. Bioessays. 1991;13:657–663. doi: 10.1002/bies.950131207. [DOI] [PubMed] [Google Scholar]
- King D.J., Cavanagh D. Infectious bronchitis. In: Calnek B.W., editor. Diseases of poultry. Ninth Edn. Iowa State University Press; Ames, IA: 1991. pp. 471–484. [Google Scholar]
- Lehrer R.I., Lichtenstein A.K., Ganz T. Defensins: antimicrobial and cytotoxic peptides of mammalian cells. Annu. Rev. Immunol. 1993;11:105–128. doi: 10.1146/annurev.iy.11.040193.000541. [DOI] [PubMed] [Google Scholar]
- Lockman H.A., Curtiss R., III Virulence of non-type 1-fimbriated and nonfimbriated nonflagellated Salmonella typhimurium mutants in murine typhoid fever. Infect. Immun. 1992;60:491–496. doi: 10.1128/iai.60.2.491-496.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MacRae E.K., Powell R.E. Cytochemical reaction for cationic proteins as a marker of primary granules during development in chick heterophils. Histochemistry. 1979;60:295–308. doi: 10.1007/BF00500657. [DOI] [PubMed] [Google Scholar]
- Muta T., Fujimoto T., Nakajima H., Iwanaga S. Tachyplesins isolated from hemocytes of southeast Asian horseshoe crabs (Carcinoscorpius rotundicauda and Tachypleus gigas): identification of a new tachyplesin, tachyplesin III, and a processing intermediate of its precursor. J. Biochem. 1990;106:261–266. doi: 10.1093/oxfordjournals.jbchem.a123191. [DOI] [PubMed] [Google Scholar]
- Ouelette A.J., Greco R.M., James M., Frederick D., Naftilan J., Fallon J.T. Developmental regulation of cryptdin, a corticostatin/defensin precursor mRNA in mouse small intestinal crypt epithelium. J. Cell. Biol. 1989;108:1687–1695. doi: 10.1083/jcb.108.5.1687. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ozaki Y., Wada K., Hase T., Matsubara H., Nakanishi T., Yoshizumi H. Amino acid sequence of a purothionin homolog from barley flour. J. Biochem. 1980;87:549–555. doi: 10.1093/oxfordjournals.jbchem.a132777. [DOI] [PubMed] [Google Scholar]
- Peck R. A one-plate assay for macrophage bactericidal activity. J. Immunol. Methods. 1985;82:131–140. doi: 10.1016/0022-1759(85)90232-7. [DOI] [PubMed] [Google Scholar]
- Penniall R., Spitznagel J.K. Vol. 72. 1975. Chicken neutrophils: oxidative metabolism in phagocytic cells devoid of myeloperoxidase; pp. 5012–5015. (Proc. Natl. Acad. Sci. USA). [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sabet T., Hsia W., Stanisz M., El-domeiri A., Van Alten P. A simple method for obtaining peritoneal macrophages from chickens. J. Immunol. Methods. 1977;14:103–110. doi: 10.1016/0022-1759(77)90001-1. [DOI] [PubMed] [Google Scholar]
- Schat K.A., Purchase H.G. Cell-culture methods. In: Purchase H.G., Arp L.H., Domermuth C.H., Pearson J.E., editors. A Laboratory Manual for the Isolation and Identification of Avian Pathogens. Third Edition. Kendall/Hunt Publishing Company; Dubuque, IA: 1989. pp. 167–175. [Google Scholar]
- Selsted M.E., Szklarek D., Lehrer R.I. Purification and antibacterial activity of antimicrobial peptides of rabbit granulocytes. Infect. Immun. 1984;45:150–154. doi: 10.1128/iai.45.1.150-154.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Selsted M.E., Szklarek D., Ganz T., Lehrer R.I. Activity of rabbit leukocyte peptides against Candida albicans. Infect. Immun. 1985;49:202–206. doi: 10.1128/iai.49.1.202-206.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Selsted M.E., Tang Y., Morris W.L., McGuire P.A., Novotny M.J., Smith W., Henschen A.H., Cullor J.S. Purification, primary structure, and antibacterial activities of beta-defensins, a new family of antimicrobial peptides from bovine neutrophils. J. Biol. Chem. 1993;268:6641–6648. [PubMed] [Google Scholar]
- Stoscheck C.M. Quantitation of Protein. In: Deutscher M.P., editor. Guide to Protein Purification. Academic Press; San Diego, CA: 1990. pp. 50–68. [Google Scholar]
- Yamashita T., Saito K. Purification, primary structure, and biological activity of guinea pig neutrophil cationic peptides. Infect. Immun. 1989;57:2405–2409. doi: 10.1128/iai.57.8.2405-2409.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zasloff M. Vol. 84. 1987. Magainins, a class of antimicrobial peptides from Xenopus skin: isolation, characterization of two active forms, and partial cDNA sequence of a precursor; pp. 5449–5453. (Proc. Natl. Acad. Sci. USA). [DOI] [PMC free article] [PubMed] [Google Scholar]