Abstract
Innate immunity is important for the integrity of the host against potentially invasive pathogenic microorganisms in the environment. Antibiotic peptides with broad antimicrobial activity are part of the innate immune system. We investigated the presence of the cathelicidin, human cationic antimicrobial protein (hCAP-18), in the male reproductive system. We found strong expression of the hCAP-18 gene by in situ hybridization and hCAP-18 protein, as detected by immunohistochemistry, in the epithelium of the epididymis, but not in the testis. The highest expression in the epididymis was in the caudal part. Western blotting showed a doublet band, the upper part corresponding to the size of hCAP-18 in plasma and neutrophils. Using a specific enzyme-linked immunosorbent assay (ELISA), levels of 86.5 +/- 37.8 microg/ml (mean +/- standard deviation; range, 41.8 to 142.8 microg/ml; n = 10) were detected in seminal plasma from healthy donors, which is 70-fold higher than the level in blood plasma. Flow cytometry and immunocytochemistry revealed the presence of hCAP-18 on spermatozoa. ELISA measurement showed levels of 196 ng/10(6) spermatozoa, corresponding to 6.6 x 10(6) molecules of hCAP-18 per spermatozoon. Our results suggest a key role for hCAP-18 in the antibacterial integrity of the male reproductive system. The attachment of hCAP-18 to spermatozoa may implicate a role for hCAP-18 in conception.
MeSH Terms
Anti-Bacterial Agents/metabolism
Antimicrobial Cationic Peptides
Carrier Proteins/metabolism
Cathelicidins
Cell Membrane/immunology,metabolism
Epididymis/immunology,metabolism
Epithelium/immunology,metabolism
Gene Expression
Humans
Immunohistochemistry
In Situ Hybridization
Male
Semen/immunology,metabolism
Spermatozoa/immunology,metabolism
Chemicals
Anti-Bacterial Agents
Antimicrobial Cationic Peptides
Carrier Proteins
Cathelicidins
ropocamptide
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Malm J
Departments of Clinical Chemistry, Malmö University Hospital, Malmö, Sweden.
[email protected]
Sørensen O
Persson T
Frohm-Nilsson M
Johansson B
Bjartell A
Lilja H
Ståhle-Bäckdahl M
Borregaard N
Egesten A
References (21)
21 references, click to expand
-
The human gene FALL39 and processing of the cathelin precursor to the antibacterial peptide LL-37 in granulocytes.
Eur J Biochem. 1996 Jun 1;238(2):325-32
PMID: 8681941
-
hCAP-18, a cathelin/pro-bactenecin-like protein of human neutrophil specific granules.
FEBS Lett. 1995 Jul 10;368(1):173-6
PMID: 7615076
-
The expression of the gene coding for the antibacterial peptide LL-37 is induced in human keratinocytes during inflammatory disorders.
J Biol Chem. 1997 Jun 13;272(24):15258-63
PMID: 9182550
-
The human antibacterial cathelicidin, hCAP-18, is synthesized in myelocytes and metamyelocytes and localized to specific granules in neutrophils.
Blood. 1997 Oct 1;90(7):2796-803
PMID: 9326247
-
An ELISA for hCAP-18, the cathelicidin present in human neutrophils and plasma.
J Immunol Methods. 1997 Aug 7;206(1-2):53-9
PMID: 9328568
-
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. 1998 Aug 4;95(16):9541-6
PMID: 9689116
-
Activities of LL-37, a cathelin-associated antimicrobial peptide of human neutrophils.
Antimicrob Agents Chemother. 1998 Sep;42(9):2206-14
PMID: 9736536
-
The human cationic antimicrobial protein (hCAP18), a peptide antibiotic, is widely expressed in human squamous epithelia and colocalizes with interleukin-6.
Infect Immun. 1999 May;67(5):2561-6
PMID: 10225921
-
The human antibacterial cathelicidin, hCAP-18, is bound to lipoproteins in plasma.
J Biol Chem. 1999 Aug 6;274(32):22445-51
PMID: 10428818
-
Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g.
Scand J Clin Lab Invest Suppl. 1968;97:77-89
PMID: 4179068
-
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature. 1970 Aug 15;227(5259):680-5
PMID: 5432063
-
Antimicrobial activity of human seminal fluid.
Scand J Urol Nephrol. 1975;9(1):17-23
PMID: 814615
-
Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4
PMID: 388439
-
Stimulus-induced maturation of probactenecins, precursors of neutrophil antimicrobial polypeptides.
J Immunol. 1991 Jun 15;146(12):4295-300
PMID: 2040802
-
The antibacterial peptide seminal plasmin alters permeability of the inner membrane of E. coli.
FEBS Lett. 1992 Jun 1;303(2-3):265-8
PMID: 1535050
-
Expression of 92-kDa type IV collagenase mRNA by eosinophils associated with basal cell carcinoma.
J Invest Dermatol. 1992 Oct;99(4):497-503
PMID: 1402008
-
Proteolytic cleavage by neutrophil elastase converts inactive storage proforms to antibacterial bactenecins.
Eur J Biochem. 1992 Oct 15;209(2):589-95
PMID: 1425666
-
A novel granulocyte-derived peptide with lipopolysaccharide-neutralizing activity.
J Immunol. 1994 Jan 1;152(1):231-40
PMID: 8254193
-
Antimicrobial activity of rabbit CAP18-derived peptides.
Antimicrob Agents Chemother. 1993 Dec;37(12):2534-9
PMID: 8109914
-
FALL-39, a putative human peptide antibiotic, is cysteine-free and expressed in bone marrow and testis.
Proc Natl Acad Sci U S A. 1995 Jan 3;92(1):195-9
PMID: 7529412
-
Widespread expression of beta-defensin hBD-1 in human secretory glands and epithelial cells.
FEBS Lett. 1996 Nov 4;396(2-3):319-22
PMID: 8915011