Evaluation of cytokine concentrations in a trehalose-stabilised lyophilised canine platelet product: a preliminary study

Background: Platelet transfusion is indicated for haemorrhage due to severe thrombocytopenia and for trauma associated coagulopathy. Febrile non-haemolytic transfusion reactions are a common complication of platelet transfusions in people and may be due to accumulated inflammatory cytokines. The present study aimed to determine the cytokine profile of a novel canine lyophilised platelet product following reconstitution, to assess the lyophilised platelets' activation response to physiological platelet agonists and to compare the cytokine profiles of basal and stimulated canine lyophilised platelets.

Methods: Cell counts and biochemical analyses were conducted following reconstitution. Cytokine concentrations were measured with a canine-specific multiplex immunocapture assay and with an electrochemiluminescent ELISA. Aliquots of reconstituted product from three separate vials were activated for 10 minutes under non-stirred conditions using adenosine diphosphate, thrombin or convulxin and their cytokine concentrations compared with unactivated samples. Flow cytometry and light-transmission aggregometry were used to evaluate the product's ability to express a procoagulant surface, degranulate and aggregate. Fresh platelet-rich plasma was used as a positive control.

Results: The product had a mean±SD particle count of 1.23±0.2×10⁹/ml, contained platelets that expressed surface phosphatidylserine before agonist stimulation and was capable of aggregation in response to thrombin stimulation suggesting that the product may have haemostatic potential following in vivo administration. Cytokine concentrations measured by the immunocapture assay were generally low, while twofold to threefold increases relative to published intervals were noted for several cytokines using the ELISA. Concentrations of chemokine (C-X-C) motif ligand 8 and tumour necrosis factor-α were significantly increased as measured by the ELISA, but not by the immunocapture assay, while concentrations of KC-like were significantly increased as measured by the immunocapture assay. Stimulation with platelet agonists did not affect measured cytokine concentrations.

Conclusion: Further study of the effects of administration of this lyophilised platelet product is warranted.

Keywords: cytokines; dogs; lyophilized; platelets; transfusion; trehalose.