Release of biologically active CD154 during collection and storage of platelet concentrates prepared for transfusion

Abstract

To cite this article: Kaufman J, Spinelli SL, Schultz E, Blumberg N, Phipps RP. Release of biologically active CD154 during collection and storage of platelet concentrates prepared for transfusion. J Thromb Haemost 2007; 5; 788-96.Summary. Background: Millions of platelet transfusions are given each year. Transfusion reactions occur in as many as 30% of patients receiving unmodified platelet transfusions. The cause of some transfusion reactions remains unclear. The current paradigm suggests that platelet concentrates (PC) contain proinflammatory mediators that are released by white blood cells during collection, processing and storage. CD154 (CD40 ligand, CD40L) is a potent inflammatory mediator, normally sequestered inside the resting platelet, that is known to translocate to the platelet membrane and be shed into plasma in response to agonist activation. We hypothesized that plateletsoluble CD154 (sCD154) is ÔspontaneouslyÕ released by transfused platelets and plays a major role in transfusion reactions. Objectives: To determine the time course and biological properties of CD154 translocation and release during collection and storage of platelets for transfusion. Methods: We measured surface and sCD154 in platelets prepared by the platelet-rich plasma method or apheresis by fluorescence-activated cell sorting and enzyme-linked immunosorbent assay, respectively. The specific biological activity of platelet sCD154 was assayed by stimulation of the CD154/CD40 pathway in known CD40-positive cells with PC-derived supernatants. Results and conclusions: We demonstrate that PCs prepared for transfusion have high levels of membrane-bound CD154 and sCD154, with maximum levels being seen 72 h after platelet collection. Importantly, we show that platelet-derived sCD154 potently stimulates CD40-positive cells. We propose that plateletderived CD154 is a key ÔcytokineÕ responsible for adverse reactions associated with platelet transfusions. Improved methods of platelet collection and/or storage, which limit CD154 expression, could reduce the risks of transfusion reaction.

Plateletpheresis: the Process, Devices, and Indicators of Product Quality

“…Various studies have investigated platelet activation, which has been used to evaluate platelet quality during processing and storage of plateletpheresies products. Activated platelet markers that have been used in studies of plateletpheresis include P-selectin (CD62P) [2,27,30], CD63 [20,22,47], gly- [12,13,19,28,62], and coated platelets [1,11,57]. Trima Accel: this suggests that increased P-selectin expression may result from delayed processing times within the apheresis device [27,30].…”

“…CD40L, a member of the tumor necrosis factor family, is a trimeric transmembrane protein that is mainly contained within platelets [12,28]. CD40L is generally sequestered inside resting platelets.…”

“…Therefore, increased sCD40L levels can provide a marker of platelet activation [12,13,19,28,62]. Platelet products that are collected with MCS+, Trima Accel, and Amicus instruments show increased sCD40L levels during storage [19,28,62].…”

Allosensitization and the Ventricular Assist Device: Dual Evolution of Technology

“…Our group has observed that soluble immune modulatory factors (sCD40L, IL-8, and RANTES) are present and biologically active in platelet concentrates [33][34][35] and non-leukoreduced red cells, and to a lesser extent this may be true of FFP as well. We hypothesize that intravenously administered blood components (including FFP), administered as a bolus (as opposed to being produced in a paracrine manner) access the lymphatic system where immune effectors reside, and modulate their responses.…”

Response: Are PDMVs the biologically active source of CD154 in ATR?

“…Indeed, recent reports indicate that acute transfusion reactions (ATRs) may result from soluble CD154 (sCD154) released from platelets during storage. 5,[8][9][10] By demonstrating that only platelet concentrates (PCs) that resulted in clinical ATR can stimulate CD154-specific B cell IL-6 production, Cognasse and colleagues make a compelling case for investigation into improved platelet storage that minimizes platelet release of CD154. Upon activation, platelets can release microparticles and exosomes 11 (collectively referred to as platelet-derived membrane vesicles [PDMVs]) that can deliver platelet-derived signals in vitro [12][13][14] or in vivo.…”