Enhanced platelet responsiveness due to chilling and its relation to CD40 ligand level and platelet–leukocyte aggregate formation

Abstract

To investigate platelet responsiveness during cold storage of whole blood, we examined platelet aggregation, expression of CD40 ligand (CD40L) on platelets, the plasma levels of soluble form of CD40L (sCD40L) as well as platelet-leukocyte aggregates. Flow cytometry analysis was performed to investigate platelet-leukocyte aggregate formation using antibodies against CD42b and CD45 and platelet activation using antibodies against P-selectin and PAC-1. Blood samples were collected from healthy volunteers, patients with cardiovascular diseases, or both. In the healthy volunteers' blood samples stored at 4 degrees C for 6 h, platelet aggregation in response to 1 micromol/l ADP was enhanced, and released levels of soluble form of P-selectin and thromboxane B2 in response to 1 micromol/l ADP markedly increased. In the samples stored at 4 degrees C for 6 h but not stimulated by any agonists, CD40L expression on the platelets was increased, and plasma levels of sCD40L were also elevated. Under the same condition, the increase in simultaneous expression of CD45 and CD42b was observed. In patients with cardiovascular diseases, the platelet aggregability, coexpression of P-selectin and PAC-1, expression of CD40L on platelets and both CD45-bound and CD42b-bound subsets were all comparable to those of healthy volunteers' samples stored at 4 degrees C for 6 h. Plasma levels of sCD40L in patients were higher than those in healthy volunteers' control. Taken together, storage of whole blood at 4 degrees C for 6 h caused platelet activation comparable to that of patients with cardiovascular diseases, and enhanced platelet activity in such patients may be involved in increased risk for thromboembolic events.

Mechanical properties and fibrin characteristics of endovascular coil–clot complexes: relevance to endovascular cerebral aneurysm repair paradigms

“…Furthermore, cellular metabolism is decreased at 5°C, reducing the formation of echinocytes. However, storage at 5°C does result in platelet activation in a manner similar to that observed in patients with cardiovascular disease 12. Although, as noted above, the major diameter of the smaller coils changed between samples, the changes were the same between each age group so that the average stiffness and fiber characteristics could be compared between age groups.…”

Release of transforming growth factor beta 1 and platelet derived growth factor type AB from canine platelet gels obtained by the tube method and activated with calcium salts

“…This methodology has been used to determine plasma TGF-β 1 concentration without interference of platelet activation (Heinemeier et al 2003). However, it is very important to consider that when cold storage or centrifugation time is prolonged platelets are activated and could develop cold storage-related lesions (Ayukawa et al 2009, Shrivastava 2009). …”

GPI-80 as a Useful Index for Myeloid Cell Heterogeneity and a Potential Prognostic Biomarker for Metastatic Renal Cell Carcinoma

“…In order to avoid neutrophil activation by the leukocyte separation, antibody reactions were performed in whole blood as described previously (Takeda et al 2014). Furthermore, it is known that platelets are activated by cold stock (4℃), which form a complex of leukocytes with platelets (Duvigneau et al 2003;Ayukawa et al 2009). Due to this, we chose a method that produces less temperature change.…”