title: Localization of radiolabeled chemotactic peptide at focal sites of Escherichia coli infection in rabbits: evidence for a receptor-specific mechanism.
abstract: The infection imaging properties of a high-affinity 99mTc-labeled chemotactic peptide receptor agonist (N-formyl-methionyl-leucyl-phenylalanine-lysine; N-For-MLFK) were compared with a low-affinity agonist (N-Acetyl-MLFK; N-Ac-MLFK), a moderate-affinity antagonist (N-isobutyloxycarbonyl-MLFK; N-IBoc-MLFK) and non-specific inflammation imaging agents.
  
  
  METHODS
  All peptides were prepared by solid-phase methods and purified by high-performance liquid chromatography. The products were assayed in vitro for N-formyl-methionyl-leucyl-phenylalanine receptor binding and superoxide production. Three types of studies were performed in rabbits with Escherichia coli infection: (Study A) Four groups of six animals were coinjected with 99mTc-N-For-MLFK-hydrazinonicotinamide (N-For-MLFK-HYNIC) plus 111In-immunoglobulin G, 111In-red blood cells or 111In-diethylene triamine pentaacetic acid. (Study B) Three groups of six rabbits were coinjected with 111In-leukocytes plus 99mTc-N-For-MLFK-HYNIC, 99mTc-N-Ac-MLFK-HYNIC or 99mTc-N-IBoc-MLFK-HYNIC. (Study C) Two groups of six rabbits were injected with 99mTc-N-For-MLFK-HYNIC and 111In-leukocytes with and without an excess of antagonist. In all three studies, the radiopharmaceuticals were injected 24 hr after infection and dual photon (99mTc and 111In) gamma camera images were acquired at 2-3 and 16-18 hr later. Target-to-background (T/B) ratios were calculated for regions of interest drawn over the infected and contralateral normal tissue.
  
  
  RESULTS
  N-For-MLFK, N-Ac-MLFK and N-IBoc-MLFK had EC50s for receptor binding of 2.0, 830 and 150 nM, respectively. The corresponding EC50s for superoxide production were 20.0, approximately 10(3) and > 10(4). Study A demonstrated that the T/B for 99mTc-N-For-MLFK-HYNIC was higher than for any of the nonspecific imaging agents (p < 0.001), and 111In-immunoglobulin G had a higher T/B ratio than 111In-diethylenetriamine pentaacetic acid (p < 0.01) or 111In-red blood cells (p = NS). Study B showed that 99mTc-N-For-MLFK-HYNIC had a higher T/B ratio than the other peptides (p < 0.001). 111In-leukocytes and 99mTc-N-IBoc-MLFK-HYNIC had comparable T/B ratios, which were higher than for 99mTc-N-Ac-MLFK-HYNIC (p < 0.05). Study C demonstrated that coinjection with an antagonist resulted in a significant reduction in the T/B ratio for 99mTc-N-For-MLFK-HYNIC (p < 0.001), but did not affect the T/B ratio for 111In-leukocytes.
  
  
  CONCLUSIONS
  Nonspecific mechanisms contribute minimally to the localization of 99mTc-chemotactic peptide analogs at sites of infection and the majority of the accumulation appears to be receptor mediated. Also, chemotactic peptide receptor antagonists can be used for infection imaging. These results provide important new insights for future radiopharmaceutical development.
authors.affiliation: Department of Radiology, Massachusetts General Hospital, Boston 02114, USA.
authors.forename: J W
authors.initials: JW
authors.lastname: Babich
authors.forename: R G
authors.initials: RG
authors.lastname: Tompkins
authors.forename: W
authors.initials: W
authors.lastname: Graham
authors.forename: S A
authors.initials: SA
authors.lastname: Barrow
authors.forename: A J
authors.initials: AJ
authors.lastname: Fischman
mesh_terms: D000818:Animals; D002630:Chemotactic Factors / Q000493:pharmacokinetics; D004912:Erythrocytes; D004927:Escherichia coli Infections / Q000000981:diagnostic imaging*; D006801:Humans; D007074:Immunoglobulin G; D007205:Indium Radioisotopes; D007962:Leukocytes; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000031:analogs & derivatives* / Q000493:pharmacokinetics; D015609:Organotechnetium Compounds / Q000493:pharmacokinetics; D004369:Pentetic Acid; D011817:Rabbits; D011877:Radionuclide Imaging; D019275:Radiopharmaceuticals / Q000493:pharmacokinetics; D018461:Soft Tissue Infections / Q000000981:diagnostic imaging*; D013667:Technetium; D013997:Time Factors; D014018:Tissue Distribution
chemical_list: D002630:Chemotactic Factors; D007074:Immunoglobulin G; D007205:Indium Radioisotopes; D015609:Organotechnetium Compounds; D019275:Radiopharmaceuticals; C107638:technetium Tc 99m N-acetylmethionyl-leucyl-phenylalanyl-lysyl-hydrazinonicotinamide; C107637:technetium Tc 99m N-isobutyloxycarbonyl-methionyl-leucyl-phenylalanyl-lysyl-hydrazinonicotinamide; C084664:technetium Tc 99m formyl-methionyl-leucyl-phenylalanyl-lysyl-hydrazinonicotinamide; D009240:N-Formylmethionine Leucyl-Phenylalanine; D013667:Technetium; D004369:Pentetic Acid; C049813:N-formylmethionyl-leucyl-phenylalanyl-lysine