Mirasol Pathogen Reduction Technology treatment does not affect acute lung injury in a two-event in vivo model caused by stored blood components

Abstract

Introduction: Mirasol Pathogen Reduction Technology (PRT) treatment uses riboflavin and UV light to inactivate pathogens in blood components. Neutrophil [polymorphonuclear cells (PMN)] priming activity accumulates during routine storage of cellular blood components, and this activity has been implicated in transfusion-related acute lung injury (TRALI). We hypothesize that PRT-treatment of blood components affects the priming activity generated during storage of packed RBCs (PRBCs) or platelet concentrates (PCs), which can elicit ALI in vivo.

Methods: Plasma, PRBCs and PCs were isolated from healthy donor's whole blood or by apheresis. Half of a collected unit was treated with PRT treatment and the remainder was left as an unmodified control. Supernatant was collected during storage of PCs and PRBCs and assayed for PMN priming activity and used as the second event in a two-event in vivo model of TRALI.

Results: PRT treatment did not induce priming activity in plasma or affect the priming activity generated during storage of PCs or PRBCs as compared with the unmodified controls. The supernatants from stored, but not fresh, PCs and PRBCs did cause ALI as the second event in a two-event animal model of TRALI, which was unaffected by PRT treatment. We conclude that the PRT treatment does not induce priming activity in plasma nor does it affect the priming activity generated during storage of PCs or PRBCs or their ability to cause ALI as the second event in a two-event in vivo model of TRALI. Moreover, the amount of priming activity in TRIMA-isolated PCs was significantly less than SPECTRA-isolated PCs.

Fig. 1

The accumulation of plasma priming activity during routine PRBC storage. Priming activity, the augmentation of the fMLP-activated respiratory burst, is depicted as a function of the storage time. The data are presented as the mean ± the standard deviation. Significant priming activity was present in both the untreated control (no treatment) and PRT-treated (treatment) PRBCs by day 14 of storage compared with supernatants from day 1 and buffer controls. Priming reached a relative maximum by day 28 of storage. PRT treatment did not increase the amount of priming activity that accumulates in the supernatant during routine PRBC storage. This figure represents the priming activity from 5 units of PRBCs from five different donors.

Fig. 2

The accumulation of plasma priming activity during routine platelet storage. Priming activity, the augmentation of the fMLP-activated respiratory burst, is shown as a function of the days of platelet storage. The data are presented as the mean ± the standard deviation. The supernatant from day 0 of both untreated (no treatment) and PRT-treated (treatment) controls did not exhibit any priming activity as compared with the buffer-treated controls. Under routine storage, PRT-treated PCs demonstrated an increase in priming activity by day 3, which was not significantly different as compared to day 0, and priming reached a relative maximum by day 7. By contrast, supernatants from the untreated control PCs did not demonstrate significant priming activity compared to day 0 until day 5 and reached a relative maximum by day 7 of storage. Importantly, there were no statistical differences between the priming activity of the PRT-treated and the untreated control units. This figure represents the data obtained from 5 double units of apheresis platelets from five disparate donors.

Fig. 3

Acute lung injury (ALI) induced by stored PRBCs is not affected by PRT treatment. Evans Blue dye (EBD) leak, a measure of pulmonary oedema, is shown as a function of treatment group. The data are represented as the mean ± SEM. In rats that received normal saline (NS) as the first event, no plasma supernatants caused ALI. However, in animals that received endotoxin (LPS) as the first event, stored (day 42) but not fresh (day 1) supernatants (non-treated) induced ALI as demonstrated by significant EBD leak from the plasma into the bronchalveolar lavage fluid (BALF) (P < 0·05). PRT treatment (treatment) did not increase ALI as quantified by EBD leak. This figure consists of a sample size of 5 with all units being represented. *P < 0·05 compared to day 1.

Fig. 4

Acute lung injury (ALI) from stored PCs in vivo is unaffected by PRT treatment. Evans blue dye (EBD) leak, a measure of pulmonary oedema, is depicted as a function of treatment group. The data are represented as the mean ± SEM. In rats that received normal saline (NS) as the first event, no supernatants caused EBD leak from the plasma into the bronchoalveolar lavage fluid (BALF). By contrast, in endotoxin- (LPS) injected rats day 5 supernatant from untreated (no treatment) and PRT-treated (treatment) stored PCs caused significant EBD leak compared with rats injected with LPS and infused with the day 0 supernatants from identical PCs (P < 0·05). PRT treatment did not increase ALI as quantified by EBD leak. This figure consists of a sample size of 5 with all 5 units being represented. *P < 0·05 compared to day 0.