Translation of glycoprotein IIIa in stored blood platelets.
BACKGROUND
Platelet (PLT) products have a short shelf life (5 days) owing in part to the deterioration of the quality of PLTs stored at 22 degrees C. This creates significant inventory challenges, and blood banks may suffer shortages and high wastage as a result. The precise biochemical pathways involved in the PLT storage lesion are unknown and must be understood before storage time can be extended.


METHODS
Informed by previous proteomics analysis, specific PLT glycoprotein (GP) concentration and surface expression were examined by Western blot and flow cytometry. mRNA concentration was determined by Northern blot and real-time polymerase chain reaction. Protein synthesis was confirmed by [(35)S]methionine labeling.


RESULTS
Western blots of GPIIIa revealed a twofold increase in concentration on Day 7 of storage and a fourfold increase on Day 10. By flow cytometry, surface expression of the GPIIb/IIIa increased by 13.4 percent on Day 7 and 41.9 percent on Day 10. Full-length GPIIIa mRNA was present throughout this storage period and was shown to have a half-life of approximately 2.9 days. Translation of GPIIb and IIIa during storage was confirmed by [(35)S]methionine labeling.


CONCLUSIONS
This article confirms that PLTs are capable of synthesizing biologically relevant proteins ex vivo throughout a 10-day storage period with particularly long-lived mRNA and provides a framework through which the biochemical mechanisms involved in the translational regulation of proteins thought to be involved in the initiation or exacerbation of the PLT storage lesion can be investigated.
Thon
Jonathan N
JN
Department of Biochemistry and Molecular Biology and the Center of Blood Research, University of British Columbia, Canada.
Devine
Dana V
DV
D001792:Blood Platelets / Q000378:metabolism*; D015152:Blotting, Northern; D015153:Blotting, Western; D005434:Flow Cytometry; D006801:Humans; D047468:Immunoprecipitation; D039661:Integrin beta3 / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013997:Time Factors
D039661:Integrin beta3; D012333:RNA, Messenger