Dekker, B.; Keen, H.; Lyons, S.; Smith, N.; Disley, L.; Hastings, D.; Williams, G.; Zweit, J.; Watson, A.
April 2003
Gut;Apr2003 Supplement 1, Vol. 52, pA105
Academic Journal
Aim and Background: We are developing molecular imaging techniques to study apoptosis (programmed cell death) in vivo directly without the need for biopsy. During apoptosis phosphatidylserine (PS) translocates from the inner to the outer leaflet of the plasma membrane. Annexin V, a protein that binds with high specificity to PS, has been shown to bind specifically to apoptotic cells in vitro and in vivo. We are investigating the use of annexin V, radiobbelled with iodine-124, as a probe for the measurement of apoptosis in vivo using PET. Methods: The probes developed in this investigation were evaluated in an in vitro (Jurkat cells were treated with camptothecin (2 µg/mL, 5 hours) and apoptosis was measured using an annexin V-FITC/propidium iodide flow cytometry assay. An in vivo model of hepatic apoptosis was developed using BDF1 mice were treated with anti-Fas (50 µg, 4 hours). Normal BDF1 mice and those treated with anti-Fas were injected with [sup 12]I--labelled proteins, and the distribution of the radiotracer measured by PET scanning. Three dimensional images were generated for analysis. Results: Significant apoptosis (p < 0.0001) was achieved in Jurkat cells after treatment with camptothecin. Eightfold increases in the radioactivity ([sup 124]I-MBP-Anx5 and [sup 124]I-annexin, but not any of the negative controls) of apoptotic cells were achieved. In normal mice, the probe was rapidly cleared, but in vivo dehalogenation resulted in the accumulation of activity in the thyroid, stomach and bladder. In mice treated with anti-Fas, radiolabelled annexin V and MBP-Anx5 (but not any of the negative control proteins) accumulated in the liver. Conclusion: MBP-Anx5 and annexin V retain their affinity for PS and apoptotic cells after direct iodination. Results of the animal experiments indicate the feasibility of using radiolabelled annexin V to detect regions of cell death in the liver in vivo.


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