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¹³¹I-Labeled Peptides as Caspase Substrates for Apoptosis Imaging

¹ Department of Radiopharmaceutical Chemistry, German Cancer Research Center, Heidelberg, Germany
² Department of Nuclear Medicine, University of Heidelberg, Heidelberg, Germany

View larger version (16K): [in a new window]   FIGURE 1. Time-course development of apoptosis obtained with camptothecin-treated Jurkat cells. (A and B) Flow cytometry using Alexa Fluor 488-labeled annexin V showing phosphatidylserine externalization (A) and formation of fluorescent rhodamine 110 mediated through caspase-3 hydrolysis of Z-DEVD-R110 (B) (n = 3). Data are expressed as mean ± SD. arb. units = arbitrary units.

View larger version (29K): [in a new window]   FIGURE 2. Propidium iodide (FL-III)/Alexa Fluor 488-annexin V (FL-I) flow cytometry of Jurkat cells treated with 10 µmol/L camptothecin for 4 h (A) and 1 µmol/L staurosporine for 3 h (B). Q1 = apoptotic cells; Q2 = apoptotic and necrotic cells; Q3 = living cells; Q4 = dead cells.

View larger version (15K): [in a new window]   FIGURE 3. Uptake kinetics of 99mTc-HYNIC-annexin V in apoptotic and control cells. Cells showed 62% apoptosis with Alexa Fluor 488-annexin V measured by flow cytometry (n = 3). Data are expressed as mean ± SD.

View larger version (16K): [in a new window]   FIGURE 4. Uptake in apoptotic and control cells of 131I-labeled DEVDGY-NH2 (1) and YDEVDG-NH2 (2) (A) as well as NQVNGY-NH2 (3) and YNQVNG-NH2 (4) (B) (n = 3). Data are expressed as mean ± SD.

View larger version (32K): [in a new window]   FIGURE 5. Uptake in apoptotic and control cells of yDEVDG-conjugated Tat sequences (5–10) (n = 3). Data are expressed as mean ± SD.

View larger version (19K): [in a new window]   FIGURE 6. Rhodamine 110 formation through caspase-mediated hydrolysis of Z-DEVD-R110 with increasing amounts of peptides 2 and 7 (n = 3). Data are expressed as mean ± SD.

View larger version (18K): [in a new window]   FIGURE 7. MALDI mass spectra of peptide 7 incubated with (A) and without (B) commercial caspase-3 (m/z represents molecular mass/charge).