title: Technetium-99m labelled antimicrobial peptides discriminate between bacterial infections and sterile inflammations.
abstract: The aim of this study was to select technetium-99m labelled peptides that can discriminate between bacterial infections and sterile inflammations. For this purpose, we first assessed the binding of various 99mTc-labelled natural or synthetic peptides, which are based on the sequence of the human antimicrobial peptide ubiquicidin (UBI) or human lactoferrin (hLF), to bacteria and to leucocytes in vitro. In order to select peptides that preferentially bind to bacteria over host cells, radiolabelled peptides were injected into mice intraperitoneally infected with Klebsiella pneumoniae (K. pneumoniae) and the amount of radioactivity associated with the bacteria and with the leucocytes was quantitated. The next phase focussed on discrimination between bacterial infections and sterile inflammatory processes using 99mTc-labelled peptides in mice intramuscularly infected with various bacteria (e.g. multi-drug-resistant Staphylococcus aureus) and in animals that had been injected with lipopolysaccharides (LPS) of bacterial origin to create a sterile inflammatory process. Also, we studied the distribution of 99mTc-labelled UBI 29-41 and UBI 18-35 in rabbits having an experimental thigh muscle infection with K. pneumoniae and in rabbits injected with LPS. Based on the results of our in vitro and in vivo binding assays, two peptides, i.e. UBI 29-41 and UBI 18-35, were selected as possible candidates for infection imaging. The radiolabelled peptides can detect infections with both gram-positive and gram-negative bacteria in mice as early as 5-30 min after injection, with a target-to-non-target (T/NT) ratio between 2 and 3; maximum T/NT ratios were seen within 1 h after injection. In rabbits, high T/NT ratios (>5) for 99mTc-labelled UBI 29-41 were observed from 1 h after injection. No accumulation of the selected 99mTc-labelled UBI-derived peptides was observed in thighs of mice and rabbits previously injected with LPS. Scintigraphic investigation into the biodistribution of 99mTc-labelled UBI peptides revealed that these peptides were rapidly removed from the circulation by renal excretion. Similar data were observed for 99mTc-labelled defensin 1-3. Our data for 99mTc-labelled hLF and related peptides indicate that these compounds are less favourable for infection detection. Taken together, 99mTc-labelled UBI 18-35 and UBI 29-41 enable discrimination between bacterial infections and sterile inflammatory processes in both mice and rabbits. Based on their characteristics, we consider these peptides the candidates of preference for detection of bacterial infections in man.
authors.affiliation: Department of Radiology, Leiden University Medical Center, The Netherlands.
authors.forename: M M
authors.initials: MM
authors.lastname: Welling
authors.forename: A
authors.initials: A
authors.lastname: Paulusma-Annema
authors.forename: H S
authors.initials: HS
authors.lastname: Balter
authors.forename: E K
authors.initials: EK
authors.lastname: Pauwels
authors.forename: P H
authors.initials: PH
authors.lastname: Nibbering
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents; D001419:Bacteria / Q000378:metabolism; D001424:Bacterial Infections / Q000000981:diagnostic imaging*; D023082:Defensins; D003937:Diagnosis, Differential; D006801:Humans; D066298:In Vitro Techniques; D007249:Inflammation / Q000139:chemically induced / Q000000981:diagnostic imaging*; D007710:Klebsiella Infections / Q000000981:diagnostic imaging; D007781:Lactoferrin; D008070:Lipopolysaccharides; D008297:Male; D051379:Mice; D011485:Protein Binding; D011506:Proteins; D011817:Rabbits; D011877:Radionuclide Imaging; D012269:Ribosomal Proteins; D013667:Technetium
chemical_list: D000900:Anti-Bacterial Agents; D023082:Defensins; D008070:Lipopolysaccharides; D011506:Proteins; D012269:Ribosomal Proteins; C040469:ribosomal protein S30; D013667:Technetium; D007781:Lactoferrin