Soluble P-selectin as Parameter for Platelet Activation during Storage

Summary

Platelet concentrates stored at room temperature deteriorate. The so-called storage lesion is characterised by morphological changes and a loss of functionality. To find an assay for early platelet activation in platelet concentrates the morphological score, β-TG release and P-selectin expression were determined, and compared with the amount of soluble P-selectin. An ELISA was used to quantify soluble P-selectin in the storage medium. We found a significant correlation between the amount of soluble P-selectin and the percentage of P-selectin positive platelets (flow-cytometric analysis) (r = 0.7449; p <0.0001) or the amount of β-TG release (r = 0.6837; p<0.0001). The morphological score also correlated significantly (negative) with the amount of soluble P-selectin (r = -0.7669; p = 0.0002). From day 0 till day 8, the amount of soluble P-selectin increased constantly from 219 ± 49.2ng/ml to 556 ± 102.3 ng/ml. The detection of soluble P-selectin can be used to quantify activation of platelets during storage. The immuno-assay for soluble P-selectin is more sensitive than flow-cytometric analysis of the percentage of P-selectin-positive cells and allows earlier detection of platelet activation.

• References

• 1 Rinder HM, Snyder EL. Activation of platelet concentrates during preparation and storage. Blood Cells 1992; 18: 445-456
• 2 Fijnheer R, Modderman PW, Veldman H, Ouwehand WH, Nieuwenhuis HK, Roos D, de Korte D. Detection of platelet activation with monoclonal antibodies and flow cytometry: changes during platelet storage. Transfusion 1990; 30: 20-25
• 3 Kunicki TJ, Tuccelli M, Becker GA, Aster RH. A study of the variables affecting the quality of platelets stored at room temperature. Transfusion 1975; 15: 414-421
• 4 Berman CL, Yeo EL, Wencel-Drake JD, Furie BC, Ginsberg MH, Furie B. A platelet alpha granule membrane protein that is associated with the plasma membrane after activation. J Clin Invest 1986; 78: 130-137
• 5 Cramer EM, Berger G, Bemdt MC. Platelet a-granule and plasma membrane share two new components: CD9 and PECAM-1. Blood 1994; 84: 1722-1730
• 6 Johnston GI, Bliss GA, Newman PJ, McEver RP. Structure of the human gene encoding granule membrane protein-140, a member of the selectin family of adhesion receptors for leukocytes. J Biol Chem 1990; 265: 21381-21385
• 7 Hamburger SA, McEver RP. GMP-140 mediates adhesion of stimulated platelets to neutrophils. Blood 1990; 75: 550-554
• 8 McEver RP. GMP-140: A receptor for neutrophils and monocytes on activated platelets and endothelium. J Cell Biochem 1991; 45: 156-161
• 9 Ishiwata N, Takio K, Katayama M, Watanabe K, Titani K, Ikeda Y, Handa M. Alternatively spliced isoform of P-selectin is present in vivo as a soluble molecule. J Biol Chem 1994; 269: 23708-23715
• 10 Dunlop LC, Skinner MP, Bendall LJ, Favaloro EJ, Castaldi PA, Gorman JJ, Gamble JR, Vadas MA, Bemdt MC. Characterization of GMP-140 (P-selectin) as a circulating plasma protein. J Exp Med 1992; 175: 1147-1150
• 11 Katayama M, Handa M, Ambo H, Araki Y, Hirai S, Kato I, Kawai Y, Watanabe K, Ikeda Y. A monoclonal antibody-based enzyme immunoassay for human GMP-140/P-selectin. J Immunol Methods 1992; 153: 41-48
• 12 Slichter SJ, Harker LA. Preparation and storage of platelet concentrates. II Storage variables influencing platelet viability and function. Br J Haematol 1976; 34: 403-419
• 13 Pietersz RNI, Loos JA, Reesink HW. Platelet concentrates stored in plasma for 72 hours at 22° C prepared from buffycoats of citrate-phosphate-dex-trose blood collected in a quadruple-bag saline-adenine-glucose-mannitol system. Vox Sang 1985; 49: 81-85
• 14 Pieters RNI, de Korte D, Reesink HW, Dekker WJA, van den Ende A, Loos JA. Storage of whole blood for up to 24 h at ambient temperature prior to component preparation. Vox Sang 1989; 56: 145-150
• 15 Loos JA. Automation in blood component preparation. In: Transfusion Medicine - Recent Technological.Advances Murawski K, Peetoom F. (eds) Alan R; Liss, Inc, New York: 1985. pp 333-341
• 16 Boomgaard MN, Gouwerok CWN, de Korte D, Loos JA. Platelets stored for 6 days in a polyolefin container in a synthetic medium with minimal plasma carry-over. Vox Sang 1994; 66: 18-24
• 17 Rinder HM, Snyder EL, Bonan JL, Napychank PA, Malkus H, Smith BR. Activation in stored platelet concentrates: correlation between membrane expression of P-selectin, glycoprotein IIb/IIIa, and beta-thromboglobulin release. Transfusion 1993; 33: 25-29
• 18 Fijnheer R, Pieters RNI, de Korte D, Gouwerok CWN, Dekker WJA, Reesink HW, Roos D. Platelet activation during preparation of platelet concentrates: A comparison of the platelet-rich plasma and the buffy coat methods. Transfusion 1990; 30: 634-638
• 19 Divers SG, Kannan K, Stewart RM, Betzing KW, Dempsey D, Fukuda M, Chervenak R, Holcombe RF. Quantitation of CD62, soluble CD62, lyso-some-associated membrane proteins 1 and 2 for evaluation of quality of stored platelet concentrates. Transfusion 1995; 35: 292-297
• 20 Wagner DD. P-selectin chases a butterfly. J Clin Invest 1995; 95: 1955-1956
• 21 Stoolman LM. Adhesion Molecules Controlling Lymphocyte Migration. Cell 1989; 56: 907-910
• 22 Goldberger A, Middleton KA, Oliver JA, Paddock C, Yan HC, Delisser HM, Albeda SM, Newman PJ. Biosynthesis and processing of the cell adhesion molecule PECAM-1 includes production of a soluble form. J Biol Chem 1994; 269: 17183-17191