- Polymorphonuclear leukocytes
- Innate immunity
- Mucosal immunity
- Fungal infections
- Bacterial infections
- Moraxella catarrhalis
- Staphylococcus aureus
- Haemophilus influenzae
- Candida albicans
Neutrophil-derived antimicrobial peptides and proteins (AMPs) play an important role in the defense against microbes. Absence of defense is illustrated by neutropenic patients with frequent bacterial and fungal infections. However, the specificity of the antimicrobial effects has not been adequately described. We set out to determine the specific antimicrobial pattern of polypeptides in neutrophils (polymorphonuclear leukocytes, PMNs) against 4 potential human pathogens: Moraxella catarrhalis, Staphylococcus aureus, Haemophilus influenzae and Candida albicans. Protein extracts of human PMNs were separated using high-performance liquid chromatography and fractions were assayed for antimicrobial activity. Fractions displaying antimicrobial activity were separated on SDS-PAGE and characterized using MALDI-MS. Depletion experiments were utilized to determine the contribution of each AMP to the antimicrobial effect. Among the identified AMPs, α-defensins 1–3, azurocidin, LL-37, lysozyme, calprotectin and lactotransferrin were studied in detail. We found a divergent pattern of killing, that is, certain peptides and proteins exhibited selective activity against specific pathogens, while others displayed a broader antimicrobial activity. α-Defensins, LL-37 and calprotectin were active against all species, while lactotransferrin exclusively inhibited growth of S. aureus. Conversely, azurocidin was active against all species except S. aureus. Our observations may shed light on bacterial resistance to AMPs and on the elimination of specific bacterial communities on mucosal surfaces.
Copyright © 2010 S. Karger AG, Basel
- Kennedy AD, DeLeo FR: Neutrophil apoptosis and the resolution of infection. Immunol Res 2009;43:25–61.
- Craig A, Mai J, Cai S, Jeyaseelan S: Neutrophil recruitment to the lungs during bacterial pneumonia. Infection and Immunity 2009;77:568–575.
- Schwartzberg LS: Neutropenia: etiology and pathogenesis. Clin Cornerstone 2006;8(suppl 5):S5–S11.
- Zeidler C, Boxer L, Dale DC, Freedman MH, Kinsey S, et al: Management of Kostmann syndrome in the G-CSF era. Br J Haematol 2000;109:490–495.
- Carlsson G, Andersson M, Putsep K, Garwicz D, Nordenskjold M, et al: Kostmann syndrome or infantile genetic agranulocytosis, part one: celebrating 50 years of clinical and basic research on severe congenital neutropenia. Acta Paediatr 2006;95:1526–1532.
- Putsep K, Carlsson G, Boman HG, Andersson M: Deficiency of antibacterial peptides in patients with morbus Kostmann: an observation study. Lancet 2002;360:1144–1149.
- Ganz T, Metcalf JA, Gallin JI, Boxer LA, Lehrer RI: Microbicidal/cytotoxic proteins of neutrophils are deficient in two disorders: Chediak-Higashi syndrome and ‘specific’ granule deficiency. J Clin Invest 1988;82:552–556.
- Ganz T: Defensins: antimicrobial peptides of innate immunity. Nat Rev Immunol 2003;3:710–720.
- Agerberth B, Gudmundsson GH: Host antimicrobial defence peptides in human disease. Curr Top Microbiol Immunol 2006;306:67–90.
- Campanelli D, Detmers PA, Nathan CF, Gabay JE: Azurocidin and a homologous serine protease from neutrophils. Differential antimicrobial and proteolytic properties. J Clin Invest 1990;85:904–915.
- Steinbakk M, Naess-Andresen CF, Lingaas E, Dale I, Brandtzaeg P, et al: Antimicrobial actions of calcium binding leucocyte L1 protein, calprotectin. Lancet 1990;336:763–765.
- Houghton AM, Hartzell WO, Robbins CS, Gomis-Ruth FX, Shapiro SD: Macrophage elastase kills bacteria within murine macrophages. Nature 2009;460:637–641.
- de Haar SF, Hiemstra PS, van Steenbergen MT, Everts V, Beertsen W: Role of polymorphonuclear leukocyte-derived serine proteinases in defense against Actinobacillus actinomycetemcomitans. Infect Immun 2006;74:5284–5291.
- Di Gennaro A, Kenne E, Wan M, Soehnlein O, Lindbom L, et al: Leukotriene B4-induced changes in vascular permeability are mediated by neutrophil release of heparin-binding protein (HBP/CAP37/azurocidin). FASEB J 2009;23:1750–1757.
- Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248–254.
- Lehrer RI, Rosenman M, Harwig SS, Jackson R, Eisenhauer P: Ultrasensitive assays for endogenous antimicrobial polypeptides. J Immunol Methods 1991;137:167–173.
- Yoshio H, Tollin M, Gudmundsson GH, Lagercrantz H, Jornvall H, et al: Antimicrobial polypeptides of human vernix caseosa and amniotic fluid: implications for newborn innate defense. Pediatr Res 2003;53:211–216.
- Turner J, Cho Y, Dinh NN, Waring AJ, Lehrer RI: Activities of LL-37, a cathelin-associated antimicrobial peptide of human neutrophils. Antimicrob Agents Chemother 1998;42:2206–2214.
- Sorensen O, Arnljots K, Cowland JB, Bainton DF, Borregaard N: The human antibacterial cathelicidin, hCAP-18, is synthesized in myelocytes and metamyelocytes and localized to specific granules in neutrophils. Blood 1997;90:2796–2803.
- Corbin BD, Seeley EH, Raab A, Feldmann J, Miller MR, et al: Metal chelation and inhibition of bacterial growth in tissue abscesses. Science 2008;319:962–965.
- Murthy AR, Lehrer RI, Harwig SS, Miyasaki KT: In vitro candidastatic properties of the human neutrophil calprotectin complex. J Immunol 1993;151:6291–6301.
- Arnold RR, Russell JE, Champion WJ, Brewer M, Gauthier JJ: Bactericidal activity of human lactoferrin: differentiation from the stasis of iron deprivation. Infect Immun 1982;35:792–799.
- Kai-Larsen Y, Bergsson G, Gudmundsson GH, Printz G, Jornvall H, et al: Antimicrobial components of the neonatal gut affected upon colonization. Pediatr Res 2007;61:530–536.
- Lee DY, Huang CM, Nakatsuji T, Thiboutot D, Kang SA, et al: Histone H4 is a major component of the antimicrobial action of human sebocytes. J Invest Dermatol 2009;129:2489–2496.
- Hirsch JG: Bactericidal action of histone. J Exp Med 1958;108:925–944.
- Bals R, Wang X, Zasloff M, Wilson JM: The peptide antibiotic LL-37/hCAP-18 is expressed in epithelia of the human lung where it has broad antimicrobial activity at the airway surface. Proc Natl Acad Sci USA 1998;95:9541–9546.
- Davis KM, Akinbi HT, Standish AJ, Weiser JN: Resistance to mucosal lysozyme compensates for the fitness deficit of peptidoglycan modifications by Streptococcus pneumoniae. PLoS Pathog 2008;4:e1000241.
- Urban CF, Ermert D, Schmid M, Abu-Abed U, Goosmann C, et al: Neutrophil extracellular traps contain calprotectin, a cytosolic protein complex involved in host defense against Candida albicans. PLoS Pathog 2009;5:e1000639.
- von Kockritz-Blickwede M, Nizet V: Innate immunity turned inside-out: antimicrobial defense by phagocyte extracellular traps. J Mol Med 2009;87:775–783.
- Lominadze G, Powell DW, Luerman GC, Link AJ, Ward RA, et al: Proteomic analysis of human neutrophil granules. Mol Cell Proteomics 2005;4:1503–1521.
- Edgeworth J, Gorman M, Bennett R, Freemont P, Hogg N: Identification of p8,14 as a highly abundant heterodimeric calcium binding protein complex of myeloid cells. J Biol Chem 1991;266:7706–7713.
- McNamara MP, Wiessner JH, Collins-Lech C, Hahn BL, Sohnle PG: Neutrophil death as a defence mechanism against Candida albicans infections. Lancet 1988;2:1163–1165.
- Aguila A, Herrera AG, Morrison D, Cosgrove B, Perojo A, et al: Bacteriostatic activity of human lactoferrin against Staphylococcus aureus is a function of its iron-binding properties and is not influenced by antibiotic resistance. FEMS Immunol Med Microbiol 2001;31:145–152.
- Lusitani D, Malawista SE, Montgomery RR: Calprotectin, an abundant cytosolic protein from human polymorphonuclear leukocytes, inhibits the growth of Borrelia burgdorferi. Infect Immun 2003;71:4711–4716.
- Clarke SR, Foster SJ: IsdA protects Staphylococcus aureus against the bactericidal protease activity of apolactoferrin. Infect Immun 2008;76:1518–1526.
- Glaser R, Harder J, Lange H, Bartels J, Christophers E, et al: Antimicrobial psoriasin (S100A7) protects human skin from Escherichia coli infection. Nat Immunol 2005;6:57–64.
- Salzman NH, Hung K, Haribhai D, Chu H, Karlsson-Sjoberg J, et al: Enteric defensins are essential regulators of intestinal microbial ecology. Nat Immunol 2010;11:76–83.
- Raqib R, Sarker P, Bergman P, Ara G, Lindh M, et al: Improved outcome in shigellosis associated with butyrate induction of an endogenous peptide antibiotic. Proc Natl Acad Sci USA 2006;103:9178–9183.
- Aujla SJ, Chan YR, Zheng M, Fei M, Askew DJ, et al: IL-22 mediates mucosal host defense against Gram-negative bacterial pneumonia. Nat Med 2008;14:275–281.
- Nograles KE ZL, Guttman-Yassky E, Fuentes-Duculan J, et al: Th17 cytokines interleukin (IL)-17 and IL-22 modulate distinct inflammatory and keratinocyte-response pathways. Br J Dermatol 2008;159:1092–1102.
- Kolls JK, McCray PB Jr, Chan YR: Cytokine-mediated regulation of antimicrobial proteins. Nat Rev Immunol 2008;8:829–835.
- Ferwerda B, Ferwerda G, Plantinga TS, Willment JA, van Spriel AB, et al: Human dectin-1 deficiency and mucocutaneous fungal infections. N Engl J Med 2009;361:1760–1767.
- Glocker EO, Hennigs A, Nabavi M, Schaffer AA, Woellner C, et al: A homozygous CARD9 mutation in a family with susceptibility to fungal infections. N Engl J Med 2009;361:1727–1735.
- Milner JD, Brenchley JM, Laurence A, Freeman AF, Hill BJ, et al: Impaired T(H)17 cell differentiation in subjects with autosomal dominant hyper-IgE syndrome. Nature 2008;452:773–776.
Dr. Peter Bergman
Department of Laboratory Medicine, Division of Clinical Microbiology F68
Karolinska Institutet and Karolinska University Hospital, Huddinge
SE–14186 Stockholm (Sweden)
Tel. +46 8 5858 0000, Fax +46 8 5858 1305, E-Mail firstname.lastname@example.org
Received: April 20, 2010
Accepted after revision: June 22, 2010
Published online: September 4, 2010
Number of Print Pages : 14
Number of Figures : 6, Number of Tables : 2, Number of References : 44
Journal of Innate Immunity
Vol. 2, No. 6, Year 2010 (Cover Date: October 2010)
Journal Editor: Herwald H. (Lund), Egesten A. (Lund)
ISSN: 1662-811X (Print), eISSN: 1662-8128 (Online)
For additional information: http://www.karger.com/JIN
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.