Relationship between the expression of GPI-anchored proteins and apoptosis in PNH erythroid precursors.

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We investigated apoptosis of erythroblasts derived from BFU-E colonies by the TUNEL method in patients with PNH and healthy volunteers. The results are as follows:1. We found that the proportion of TUNEL-positive erythroblasts was low in PNH patients and healthy volunteers, and that there were no significant differences between them (Shichishima et al, Blood, 1997).2. The fact described above suggests that apoptosis of cultured erythroblasts is not associated with the expression of glycosylphosphatidylinositol (GPI)-anchored membrane proteins in PNH.Recently, it has been reported in some papers that peripheral blood granulocytes derived from PNH patients are resistant to apoptosis compared with those from healthy volunteers.Accordingly, we also investigated apoptosis of peripheral blood granulocytes by the methods using Annexin V expression and 7-AAD in PNH patients and healthy volunteers. The results are as follows:1. We found that each of the granulocyte number after 0, 6, 12, 18, and 24 hours during culture in serum-free medium was similar between PNH patients and healthy volunteers.2. We were not able to find no significant differences between the apoptotic granulocyte number from PNH patients and that from healthy volunteers during 0, 6, 12, 18, and 24 hour culture in serum-free medium by the methods using Annexin V expression and 7-AAD.3. We confirmed that peripheral blood granulocytes derived from patients with myelodysplatic syndrome were resistant to apoptosis compared with those from both PNH patients and healthy volunteers by the methods using Annexin V expression and 7-AAD.The findings described in Items 1 to 3 suggest that caspase-3 activity and CD95 expression of the granulocytes from PNH patients should be investigated in the near future. After the study, we are going to submit our paper to any journal for hematology.