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A new 99mTc labelling method for leucocytes: in vitro and in vivo comparison with 99mTc-HMPAO.

Abstract
A new method for the labelling of mixed leucocytes with 99mTc-tropolone was optimized and compared with a 99mTc-HMPAO leucocyte labelling procedure in vitro and in vivo. In the present study, leucocytes obtained from patients suffering from Crohn's disease, were isolated and labelled with 99mTc-HMPAO or labelled according the new 99mTc-tropolone procedure using 9.8 mM tropolone, 1 microM stannous chloride and 0.8 mM potassium borohydride (KBH4) at pH 5.5-6. Labelling efficiency with 99mTc-tropolone yielded 92 +/- 3%, which is higher compared to the 99mTc-HMPAO labelling procedure (64 +/- 13%) using 10(8) of leucocytes. In vitro stability and viability of both the tropolone and the HMPAO labelled cells was investigated. The viability test of the 99mTc-labelled leucocytes was performed in autologous plasma at 37 degrees C and compared with unlabelled leucocytes. After 18 hours of incubation a significant (P < 0.05) higher stability was observed for 99mTc-tropolone labelled leucocytes (84 +/- 5%) compared with that of 99mTc-HMPAO labelled leucocytes (73 +/- 5%). The viability of the 99mTc-labelled leucocytes observed for both labelling procedures was similar to unlabelled leucocytes. In vivo experiments were performed in mice. 99mTc-tropolone or 99mTc-HMPAO labelled murine mixed leucocytes were injected in mice, with a Staphylococcus aureus ATCC 25923 thigh infection. Analysis of scintigraphic images yielded a faster clearance of the 99mTc-tropolone labelled leucocytes. This was most likely due to a significant (P < 0.02) higher liver uptake at 4 hours after administration of the 99mTc-tropolone labelled leucocytes (19%) in comparison with 99mTc-HMPAO labelled cells (9%). Faster and significant (P < 0.02) higher accumulation of the 99mTc-tropolone labelled leucocytes was observed at the site of infection compared with 99mTc-HMPAO labelled leucocytes at all time-intervals after the administration of the 99mTc-labelled leucocytes. The new 99mTc-tropolone leucocyte labelling procedure, offers an attractive low-cost agent for research purposes.
AuthorsM Welling, H I Feitsma, D Blok, W Calame, G J Ensing, W Goedemans, E K Pauwels
JournalThe quarterly journal of nuclear medicine : official publication of the Italian Association of Nuclear Medicine (AIMN) [and] the International Association of Radiopharmacology (IAR) (Q J Nucl Med) Vol. 39 Issue 2 Pg. 89-98 (Jun 1995) ISSN: 1125-0135 [Print] ITALY
PMID8574810 (Publication Type: Comparative Study, Journal Article)
Chemical References
  • Borohydrides
  • Organotechnetium Compounds
  • Oximes
  • Tin Compounds
  • potassium borohydride
  • stannous chloride
  • Technetium Tc 99m Exametazime
  • Tropolone
Topics
  • Abscess (radionuclide imaging)
  • Animals
  • Borohydrides
  • Cell Survival
  • Chromatography
  • Humans
  • Hydrogen-Ion Concentration
  • In Vitro Techniques
  • Isotope Labeling
  • Leukocytes
  • Male
  • Mice
  • Organotechnetium Compounds
  • Oximes
  • Staphylococcal Infections (radionuclide imaging)
  • Technetium Tc 99m Exametazime
  • Tin Compounds
  • Tropolone

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