On the wave of the encouraging results showed by Z. DARZYNKIEWICZ et al. in leukemia patients we tried to recognize apoptosis in vivo in circulating blasts by using TUNEL in patients affected by leukemias with high number of cells during chemotherapy. Although we were able to detect apoptotic circulating blasts, the maximum number of TUNEL positivecells was 3% even in patients with a rapid decrease in circulating blasts. We studied these patient for several days every 24 hours until aplasia was reached. The results were "disappointing" and the question which arise is: Is the number of circulating apoptotic cells just a matter of induction of apoptosis, or of the effectiveness of the hystiocytes/macrophage tissues which remove those cells from circulation? Probably both! Could we consider these two phenomena simultaneously? Probably not! It's better to work with solid tissues were apoptotic cells stay longer. (See Raza et al.) Good luck. Ugo Consoli, M.D. Institute of Hematology University of Catania, ITALY Fax: ++39/95-7311510 Tel: ++39/95-7435917 ---------------------------------------------------------------------------- ----------------------------------------------- Matthias Haury wrote: Salut tout le monde... Does anybody of you have experience of using the TUNEL technique to test apoptosis in mouse thymocytes directly ex vivo (no incubation in culture) ? I'm just curious if one can detect the apopotosis of double positive thymocytes directly, or if a incubation in culture is necessary to see anything... I know that this subject has been discussed before, but it's very hard to search the archive from here, as the network connections are terribly jammed in the moment... Thanks a lot again, Matthias _____________________________________________________________________________ Matthias Haury Flowcytometry Dept Immunology Institut Pasteur mhaury@pasteur.fr Tel: 33 (01) 40 61 31 29 Fax: 33 (01) 45 68 86 39 _____________________________________________________________________________
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