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Apoptotic markers are increased in platelets stored at 37°C
Author(s) -
Bertino A.M.,
Qi X.Q.,
Li J.,
Xia Y.,
Kuter D.J.
Publication year - 2003
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1046/j.1537-2995.2003.t01-4-00431.x
Subject(s) - apoptosis , gelsolin , platelet , viability assay , chemistry , blot , caspase 3 , cleavage (geology) , caspase , andrology , microbiology and biotechnology , biochemistry , programmed cell death , biology , immunology , medicine , paleontology , actin , fracture (geology) , gene
BACKGROUND: PLTs for transfusion lose viability during storage in blood banking. This loss of viability is accelerated at 37°C, as is the risk of bacterial contamination, and has led to the selection of 22°C as the routine storage temperature. Because PLTs contain an intact apoptotic mechanism, we sought to determine whether PLTs undergo apoptosis during storage and whether storage at 37°C accelerated this process. STUDY DESIGN AND METHODS: PLT‐rich plasma from PLT concentrates was stored at 37 or 22°C in small aliquots or whole bags, with and without cell‐permeable caspase inhibitors. Number of PLTs, pH, LDH level, and 3‐(4,5‐dimethylthiazol‐2‐yl)‐5‐(3‐carboxymethoxyphenyl)‐2‐(4‐sulfophenyl)‐2H‐tetrazolium activity were analyzed over time. PLT lysates were prepared and tested for the presence and activation of apoptotic proteins by enzyme assay and Western blotting. RESULTS: PLT viability was greatly reduced after 1 to 2 days of storage at 37°C; however, signs of apoptosis were evident by 3 hours after temperature shift. In temperature‐stressed PLTs only, a gradual rise in caspase‐3 activity was detected that correlated with the appearance of the 17‐ to 20‐kDa cleavage products of caspase‐3. Gelsolin, a caspase‐3 substrate, underwent cleavage within the same time frame. Bcl‐x L and caspase‐2 also declined significantly; caspase‐9 activity rose. Specific caspase inhibitors could prevent caspase activation but did not improve PLT cellular viability at 37°C. CONCLUSIONS: PLTs contain apoptotic proteins that are activated during PLT storage at 37°C and may account for the rapid decline in PLT cellular viability. Although ineffective here, inhibition of PLT apoptosis may improve PLT cellular viability.