99mTc-labeled antimicrobial peptides for detection of bacterial and Candida albicans infections.
This study compared the possibilities and limitations of 99mTc-labeled synthetic peptides derived from two human antimicrobial peptides, namely, ubiquicidin (UBI) and lactoferrin (hLF), for the scintigraphic detection of bacterial and fungal infections in mice and rabbits. The rationale of our approach was that selected peptides accumulate in infected areas but not in sterile inflammatory lesions, because they bind preferentially to microorganisms. 99mTc-labeled human neutrophil peptides (defensins), ciprofloxacin, and human polyclonal IgG were included as control agents.


METHODS
99mTc-labeled peptides and control agents were injected intravenously into animals that had been injected intramuscularly 18 h earlier with multidrug-resistant Staphylococcus aureus, Klebsiella pneumoniae, or fluconazole-resistant Candida albicans. Sterile inflammatory sites were induced by the injection of heat-killed microorganisms or lipopolysaccharide (LPS) into the thigh muscle. Up to 4 h after injection, the accumulation of 99mTc-labeled compounds in the infected/inflamed thigh muscles was determined using scintigraphic techniques and radioactivity counts in dissected tissues.


RESULTS
Scintigraphy revealed that 99mTc-labeled peptides UBI 29-41, UBI 18-35, UBI 31-38, hLF 1-11, and defensins, which showed preferential in vitro binding to microorganisms in a former study, accumulated at a significantly higher rate (P < 0.01) in bacterial and C. albicans infections in mice and rabbits than in inflamed tissues induced by heat-killed microorganisms or by LPS. No significant difference in the accumulation of 99mTc-labeled ciprofloxacin was observed between infected and sterile inflamed thigh muscles in mice.


CONCLUSIONS
99mTc-labeled antimicrobial peptides UBI 29-41, UBI 18-35, UBI 31-38, hLF 1-11, and defensins accumulate significantly in tissues infected with gram-positive and gram-negative bacteria and C. albicans. Significantly lower (P < 0.01) accumulation of these peptides occurs in sterile inflamed tissues. These data indicate that the peptides preferentially tag microorganisms at the site of infection, which is in agreement with their preferential binding to the microorganisms in vitro and in vivo. 99mTc-labeled ciprofloxacin does not distinguish between infections and sterile inflammatory lesions, which implies that its specificity for the detection of bacterial infections is not warranted.
Welling
M M
MM
Department of Radiology, Leiden University Medical Center, Leiden, The Netherlands.
Lupetti
A
A
Balter
H S
HS
Lanzzeri
S
S
Souto
B
B
Rey
A M
AM
Savio
E O
EO
Paulusma-Annema
A
A
Pauwels
E K
EK
Nibbering
P H
PH
D000818:Animals; D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; D001424:Bacterial Infections / Q000000981:diagnostic imaging*; D002177:Candidiasis / Q000000981:diagnostic imaging*; D002939:Ciprofloxacin; D023082:Defensins; D018432:Drug Resistance, Multiple; D007074:Immunoglobulin G; D007249:Inflammation; D007710:Klebsiella Infections / Q000175:diagnosis; D007781:Lactoferrin; D008297:Male; D051379:Mice; D011817:Rabbits; D011877:Radionuclide Imaging; D019275:Radiopharmaceuticals; D012269:Ribosomal Proteins; D013203:Staphylococcal Infections / Q000000981:diagnostic imaging; D013211:Staphylococcus aureus / Q000187:drug effects; D013667:Technetium
D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; D023082:Defensins; D007074:Immunoglobulin G; D019275:Radiopharmaceuticals; D012269:Ribosomal Proteins; C040469:ribosomal protein S30; D002939:Ciprofloxacin; D013667:Technetium; D007781:Lactoferrin