title: Development of a Hybrid Tracer for SPECT and Optical Imaging of Bacterial Infections.
abstract: In trauma and orthopedic surgery, infection of implants has a major impact on the outcome for patients. Infections may develop either during the initial implantation or during the lifetime of an implant. Both infections, as well as aseptic loosening of the implant, are reasons for revision of the implants. Therefore, discrimination between aseptic-mechanical-loosening and septic-bacterial-loosening of implants is critical during selection of a patient-tailored treatment policy. Specific detection and visualization of infections is a challenge because it is difficult to discriminate infections from inflammation. An imaging tracer that facilitates bacterial identification in a pre- and intraoperative setting may aid the workup for patients suspicious of bacterial infections. In this study we evaluated an antimicrobial peptide conjugated to a hybrid label, which contains both a radioisotope and a fluorescent dye. After synthesis of DTPA-Cy5-UBI29-41 and-when necessary-radiolabeling with (111)In (yield 96.3 ± 2.7%), in vitro binding to various bacterial strains was evaluated using radioactivity counting and confocal fluorescence microscopy. Intramuscular bacterial infections (S. aureus or K. pneumoniae) were also visualized in vivo using a combined nuclear and fluorescence imaging system. The indium-111 was chosen as label as it has a well-defined coordination chemistry, and in pilot studies labeling DTPA-Cy5-UBI29-41 with technetium-99m, we encountered damage to the Cy5 dye after the reduction with SnCl2. As a reference, we used the validated tracer (99m)Tc-UBI29-41. Fast renal excretion of (111)In-DTPA-Cy5-UBI29-41 was observed. Target to nontarget (T/NT) ratios were highest at 2 h post injection: radioactivity counting yielded T/NT ratios of 2.82 ± 0.32 for S. aureus and 2.37 ± 0.05 for K. pneumoniae. Comparable T/NT ratios with fluorescence imaging of 2.38 ± 0.09 for S. aureus and 3.55 ± 0.31 for K. pneumoniae were calculated. Ex vivo confocal microscopy of excised infected tissues showed specific binding of the tracer to bacteria. Using a combination of nuclear and fluorescence imaging techniques, the hybrid antimicrobial peptide conjugate DTPA-Cy5-UBI29-41 was shown to specifically accumulate in bacterial infections. This hybrid tracer may facilitate integration of noninvasive identification of infections and their extent as well as real-time fluorescence guidance during surgical resection of infected areas.
authors: Mick M
authors: MM
authors: Welling
authors: Anton
authors: A
authors: Bunschoten
authors: Joeri
authors: J
authors: Kuil
authors: Rob G H H
authors: RG
authors: Nelissen
authors: §Delft University of Technology, 2628 CD Delft, The Netherlands.|∥MILabs, 3584 CX Utrecht, The Netherlands.
authors: Freek J
authors: FJ
authors: Beekman
authors: Tessa
authors: T
authors: Buckle
authors: Fijs W B
authors: FW
authors: van Leeuwen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002232:Carbocyanines / Q000737:chemistry; D002460:Cell Line; D004396:Coloring Agents / Q000737:chemistry; D006801:Humans; D007205:Indium Radioisotopes; D007710:Klebsiella Infections / Q000000981:diagnostic imaging*; D007711:Klebsiella pneumoniae / Q000502:physiology; D051379:Mice; D061848:Optical Imaging / Q000379:methods*; D004369:Pentetic Acid / Q000737:chemistry; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry* / Q000493:pharmacokinetics / Q000633:toxicity; D011849:Radioactive Tracers; D012269:Ribosomal Proteins / Q000737:chemistry; D013203:Staphylococcal Infections / Q000000981:diagnostic imaging*; D013211:Staphylococcus aureus / Q000502:physiology; D015899:Tomography, Emission-Computed, Single-Photon / Q000379:methods*
chemical_list: D002232:Carbocyanines; D004396:Coloring Agents; D007205:Indium Radioisotopes; D010446:Peptide Fragments; D011849:Radioactive Tracers; D012269:Ribosomal Proteins; C085321:cyanine dye 5; C040469:ribosomal protein S30; D004369:Pentetic Acid