Localization and stability of technetium-99m-Sn-pyrophosphate in rat neutrophils

Technetium-99m has been suggested as an alternative radiolabel for white cells, and while its physical characteristics are nearly ideal, its stability and site of localization in this procedure are unclear. We examined these parameters by radiolabeling 10(8) neutrophils from rat peritoneum with 74 to 370 MBq technetium-99m-Sn-pyrophosphate. We found that the percentage of initial activity bound to neutrophils was quite variable, possibly because the radiolabel associated with several subfractions: 19.8 +/- 11.5% (mean +/- s.d.) with nuclei and plasma membranes, 25.6 +/- 3.9% with mitochondria, 26.6 +/- 9.8% with microsomes, and 29.2 +/- 6.9% with cytosol. Approximately 80-90% of the radioactivity associated with neutrophils was not bound to protein and only about one-half of the activity localized to cell membranes was removable over 4 hr by pepsin digestion. We concluded that the variable labeling efficiency was due to the radiolabel's rather loose association with several cellular subfractions rather than specific binding to a unique substrate.