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Translation of glycoprotein IIIa in stored blood platelets

Authors

  • Jonathan N. Thon,

    1. From the Department of Biochemistry and Molecular Biology and the Center of Blood Research, University of British Columbia, and Canadian Blood Services, Vancouver, British Columbia, Canada.
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  • Dana V. Devine

    1. From the Department of Biochemistry and Molecular Biology and the Center of Blood Research, University of British Columbia, and Canadian Blood Services, Vancouver, British Columbia, Canada.
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  • This work is supported by the Canadian Blood Services. JNT is supported by a Canadian Blood Services graduate fellowship program and by a CIHR graduate fellowship in transfusion science.

Dana Devine, Canadian Blood Services, UBC Center for Blood Research, 4th Floor, 2350 Health Sciences Mall, Vancouver, BC, V6T 1Z3, Canada. e-mail: dana.devine@blood.ca.

Abstract

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°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.

STUDY DESIGN AND 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 [35S]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 [35S]methionine labeling.

CONCLUSION: 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.

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