The human antimicrobial and chemotactic peptides LL-37 and α-defensins are expressed by specific lymphocyte and monocyte populations

Blood - Tập 96 - Trang 3086-3093 - 2000
Birgitta Agerberth, Jehad Charo, Joachim Werr, Berit Olsson, Farah Idali, Lennart Lindbom, Rolf Kiessling, Hans Jörnvall, Hans Wigzell, Gudmundur H. Gudmundsson

Tóm tắt

AbstractWe identified antibacterial components in human T and natural killer (NK) cells by using freshly isolated lymphocytes enriched for T and NK cells as starting material. After growing these lymphocytes for 5 days in the presence of interleukin (IL)–2, we isolated and characterized several antibacterial peptides/proteins from the supernatant—α-defensins (HNP 1-3), LL-37, lysozyme, and a fragment of histone H2B—although other active components were also present. We then used reverse transcriptase–polymerase chain reaction to search for expression of the gene coding for LL-37 in several B-cell lines, γδ T-cell lines, NK clones, and one monocytic cell line, with positive results, but found no expression in several αβ T-cell lines. The α-defensins (HNP 1-3) were also found to be expressed in several of these cell lines. To confirm the presence of these antibacterial peptides in lymphocytes, we localized them to NK, γδ T cells, B cells, and monocytes/macrophages by using double-staining immunohistochemical analysis of freshly isolated lymphocytes. We also found that primary cultures of lymphocytes transcribe and secrete LL-37 and that these processes are affected by IL-6 and interferon-γ. In addition, we demonstrated that LL-37 has chemotactic activity for polymorphonuclear leukocytes and CD4 T lymphocytes, whereas others have shown chemotactic activity for human α-defensins (HNP 1-2). These findings suggest that microbicidal peptides are effector molecules of lymphocytes and that antibacterial activity previously shown to be derived from T and NK cells may be partly mediated by the antibacterial peptides LL-37 and HNP 1-3.

Tài liệu tham khảo

Stolzenberg, 1997, Epithelial antibiotic induced in states of disease., Proc Natl Acad Sci U S A., 94, 8686, 10.1073/pnas.94.16.8686 Frohm, 1997, The expression of the gene coding for the antibacterial peptide LL-37 is induced in human keratinocytes during inflammatory disorders., J Biol Chem., 272, 15258, 10.1074/jbc.272.24.15258 Sorensen, 1997, An ELISA for hCAP-18, the cathelicidin present in human neutrophils and plasma., J Immunol Methods., 206, 53, 10.1016/S0022-1759(97)00084-7 Lehrer, 1993, Defensins: antimicrobial and cytotoxic peptides of mammalian cells., Annu Rev Immunol., 11, 105, 10.1146/annurev.iy.11.040193.000541 Jones, 1992, Paneth cells of the human small intestine express an antimicrobial peptide gene., J Biol Chem., 267, 23216, 10.1016/S0021-9258(18)50079-X Jones, 1993, Defensin-6 mRNA in human Paneth cells: implications for antimicrobial peptides in host defense of the human bowel., FEBS Lett., 315, 187, 10.1016/0014-5793(93)81160-2 Zhao, 1996, Widespread expression of beta-defensin hBD-1 in human secretory glands and epithelial cells., FEBS Lett., 396, 319, 10.1016/0014-5793(96)01123-4 Harder, 1997, A peptide antibiotic from human skin., Nature., 387, 861, 10.1038/43088 Gudmundsson, 1996, The human gene FALL39 and processing of the cathelin precursor to the antibacterial peptide LL-37 in granulocytes., Eur J Biochem., 238, 325, 10.1111/j.1432-1033.1996.0325z.x Bals, 1998, 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 U S A., 95, 9541, 10.1073/pnas.95.16.9541 Agerberth, 1999, Antibacterial components in bronchoalveolar lavage fluid from healthy individuals and sarcoidosis patients., Am J Respir Crit Care Med., 160, 283, 10.1164/ajrccm.160.1.9807041 Zanetti, 1995, Cathelicidins: a novel protein family with a common proregion and a variable C-terminal antimicrobial domain., FEBS Lett., 374, 1, 10.1016/0014-5793(95)01050-O Ganz, 1997, Antimicrobial peptides of phagocytes and epithelia., Semin Hematol., 34, 343 Smith, 1996, Cystic fibrosis airway epithelia fail to kill bacteria because of abnormal airway surface fluid., Cell., 85, 229, 10.1016/S0092-8674(00)81099-5 Wilson, 1999, Regulation of intestinal α-defensin activation by the metalloproteinase matrilysin in innate host defense., Science., 286, 113, 10.1126/science.286.5437.113 Chertov, 1996, Identification of defensin-1, defensin-2, and CAP37/azurocidin as T-cell chemoattractant proteins released from interleukin-8-stimulated neutrophils., J Biol Chem., 271, 2935, 10.1074/jbc.271.6.2935 Dempsey, 1996, C3d of complement as a molecular adjuvant: bridging innate and acquired immunity., Science., 271, 348, 10.1126/science.271.5247.348 Yang, 1999, β-Defensins: linking innate and adaptive immunity through dendritic and T cell CCR6., Science., 286, 525, 10.1126/science.286.5439.525 Garcia-Penarrubia, 1989, Antibacterial activity of human natural killer cells., J Exp Med., 169, 99, 10.1084/jem.169.1.99 Levitz, 1995, Direct antimicrobial activity of T cells., Immunol Today., 16, 387, 10.1016/0167-5699(95)80007-7 Andersson, 1995, NK-lysin, a novel effector peptide of cytotoxic T and NK cells. Structure and cDNA cloning of the porcine form, induction by interleukin 2, antibacterial and antitumour activity., EMBO J., 14, 1615, 10.1002/j.1460-2075.1995.tb07150.x Stenger, 1998, An antimicrobial activity of cytolytic T cells mediated by granulysin., Science., 282, 121, 10.1126/science.282.5386.121 Agerberth, 1995, FALL-39, a putative human peptide antibiotic, is cysteine-free and expressed in bone marrow and testis., Proc Natl Acad Sci U S A., 92, 195, 10.1073/pnas.92.1.195 Burnette, 1981, “Western blotting”: electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A., Anal Biochem., 112, 195, 10.1016/0003-2697(81)90281-5 Sambrook, 1989, Molecular Cloning: A Laboratory Manual., 2nd ed. Frohman, 1988, Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer., Proc Natl Acad Sci U S A., 85, 8998, 10.1073/pnas.85.23.8998 Johansson, 1998, Conformation-dependent antibacterial activity of the naturally occurring human peptide LL-37., J Biol Chem., 273, 3718, 10.1074/jbc.273.6.3718 Czarniecki, 1993, Interferon-γ and resistance to bacterial infections., APMIS., 101, 1, 10.1111/j.1699-0463.1993.tb00073.x Prokesova, 1994, Antibacterial activity of human mononuclear leukocytes against Staphylococcus aureus., Folia Microbiol (Praha)., 39, 428, 10.1007/BF02814451 Garcia-Penarrubia, 1989, Experimental and theoretical kinetics study of antibacterial killing mediated by human natural killer cells., J Immunol., 142, 1310, 10.4049/jimmunol.142.4.1310 Hiemstra, 1993, Antimicrobial proteins of murine macrophages., Infect Immun., 61, 3038, 10.1128/IAI.61.7.3038-3046.1993 Rose, 1998, Potential role of epithelial cell-derived histone H1 proteins in innate antimicrobial defense in the human gastrointestinal tract., Infect Immun., 66, 3255, 10.1128/IAI.66.7.3255-3263.1998 Schroder, 1999, Epithelial peptide antibiotics., Biochem Pharmacol., 57, 121, 10.1016/S0006-2952(98)00226-3 Akira, 1995, Targeted disruption of the IL-6 related genes: gp130 and NF-IL-6., Immunol Rev., 148, 221, 10.1111/j.1600-065X.1995.tb00100.x Rottenberg, 1999, Role of innate and adaptive immunity in the outcome of primary infection with Chlamydia pneumoniae, as analyzed in genetically modified mice., J Immunol., 162, 2829, 10.4049/jimmunol.162.5.2829 Murray, 1999, Defining the requirements for immunological control of mycobacterial infections., Trends Microbiol., 7, 366, 10.1016/S0966-842X(99)01567-X Andreu, 1998, Animal antimicrobial peptides: an overview., Biopolymers., 47, 415, 10.1002/(SICI)1097-0282(1998)47:6<415::AID-BIP2>3.0.CO;2-D Lillard, 1999, Mechanisms for induction of acquired host immunity by neutrophil peptide defensins., Proc Natl Acad Sci U S A., 96, 651, 10.1073/pnas.96.2.651 Gudmundsson, 1999, Neutrophil antibacterial peptides, multifunctional effector molecules in the mammalian immune system., J Immunol Methods., 232, 45, 10.1016/S0022-1759(99)00152-0