Vox Sanguinis (2015) 108, 205–207 © 2014 International Society of Blood Transfusion DOI: 10.1111/vox.12203
SHORT REPORT
Amotosalen-HCl-UVA pathogen reduction does not alter poststorage metabolism of soluble CD40 ligand, Ox40 ligand and interkeukin-27, the cytokines that generally associate with serious adverse events H. Hamzeh-Cognasse,1 S. Laradi,1,2 J. C. Osselaer,3 F. Cognasse1,2 & O. Garraud1,4 1
Universite de Lyon, GIMAP-EA3064, Saint Etienne, France EFS Auvergne-Loire, Saint Etienne, France 3 Service re gional vaudois de transfusion sanguine, Epalinges, Switzerland 4 Institut National de Transfusion Sanguine (INTS), Paris, France 2
Received: 6 July 2014, revised 28 August 2014, accepted 5 September 2014, published online 21 October 2014
Platelets in therapeutic platelet concentrates are commonly acknowledged to release biologically active constituents during storage. This study examined the influence of photochemical pathogen reduction treatment (PRT) using amotosalen-HCl and UVA light vs. untreated control platelet components, on three factors recently reported to be associated with serious adverse events associated with platelet component (PC) transfusions: sCD40L, IL-27 and sOX40 ligand. Levels of such cytokine-like factors increased significantly during storage, but no significant difference was detected between PRT- and control PCs. This suggests that occurrences of AEs are not directly influenced by PRT but rather may depend on alternate determinants. Key words: cytokine, inflammation, pathogen reduction treatment, platelet storage.
Introduction Pathogen reduction technologies (PRTs) are applicable to platelet components (PCs) with the aim of providing three main benefits: eradication of bacterial contaminations, reduction/inactivation of most known and emergent viruses and inactivation of residual leucocytes. Over a near 10-year experience with this process in some blood establishments (BEs), the Intercept system (amotosalen/HCl/UVA illumination) proved successful on all three targets [1]. PRTs have not been implemented universally yet, mainly because of cost impact, loss of platelets in the PCs and residual concerns on platelet function alteration. It remains relevant to examine whether there is a variation in the platelet capacity for secretion of cytokines and other biological response modifiers (BRMs) that are characteristic of Correspondence: Olivier Garraud, INTS and GIMAP-EA 3064, Universite de Saint-Etienne, Faculte de Medecine, 15 rue Ambroise Pare, 42023 Saint-Etienne Cedex 2, France E-mail: [email protected]
platelet physiology and also present in PCs associated with grade 2–3 serious adverse events, and principally those that are inflammatory in nature [2]. It is worth noticing that reduction of inflammation, suggested as a potential benefit by manufacturers, is not the primary goal of PRTs [3]. Because of novel evidence that certain cytokine-like BRMs are specifically associated with AEs related to PC transfusions, namely sOx40L and IL-27 (along with sCD40L) [4], we aimed at re-evaluating the potential of amotosalenUVA treatment to impact (increase/decrease vs. steady state conditions) BRMs during preparation and storage of PCs, with shelf-life extended to 7 days.
Materials and methods Blood collection using platelet apheresis and PRT were performed (n = 15) as reported previously [5]. Quality control parameters were as follows: mean volume (419 – 30 ml); mean platelet count (5.6 – 0.6 9 1011 platelets/component); mean residual leucocyte count (0.095 – 0.087 9 106 leucocytes/component); and mean 205
206 H. Hamzeh-Cognasse et al.
pH (7.3 – 0.1; d1). All PCs were automatically resuspended in nominal 35% donor plasma and 65% platelet additive solution (InterSol, Fenwal-Europe, Belgium) and divided into paired units (n = 15) of equal volume and platelet content. One component was retained as an untreated control, and the other treated with PRT with 150 lmol/l amotosalen, 3 J/cm2 UVA treatment and CAD absorption). Platelet concentration (106/ml), pH and levels of sCD40L, IL-27 and sOX40L were measured on Days 0–7. Endotoxin levels, sCD40L, IL-27 and sOX40L were measured as described previously and adjusted to 109 platelets [4]. Differences between cytokine levels over time were evaluated by one-way analysis of variance (ANOVA) followed by Bonferroni’s correction. As values were not normally distributed, Interexperiment comparisons of marker expression and cytokine and/or chemokine concentrations were analysed with the nonparametric analysis of variance with the Kruskal–Wallis test using STATVIEW (SAS Institute, Cary, NC, USA) and P values of 0.05) from untreated PCs during 7 days of storage. From Day 1 to 7, the pH of PRTand control PCs decreased similarly, from 7.1 – 0.1 in both D1, to 6.9 – 0.1 and 6.8 – 0.1, respectively, on D7 (data not shown), but remained within acceptable ranges for therapeutic transfusion. IL-27 secretion by platelets in untreated PCs was similar to the sCD40L secretion pattern overtime as previously observed (Fig. 1c), [4] and with a significant increase by day 3 (28600 – 1872 pg/ml *P < 0.05) and a twofold amount of this cytokine on days 5 and 7 (37921 – 2071 and 41234 – 1004 pg/ml *P < 0.05) (Fig. 1a). Soluble (s)OX40L secretion was objectively stable in untreated PCs until day 5 but increased when PCs were stored for another 2 days (d7: 864 – 27 pg/ml, compared to d0: 382 – 12 pg/ml *P < 0.05; Fig. 1b). No differences were detected, however, between PRT- and control PCs regarding all three BRMs, at all times of storage. However, there was a strong trend, concerning elevated IL-27 secretion by platelets in PRT- vs. control PCs, from day 2 of storage (d2, P = 0.066; d3, P = 0.11; d4, P = 0.12; d5, P = 0.075). Several investigators reported that PRT variably affect the levels of platelet-derived cytokines in PCs [6]. Recent investigations, however, show that PRT-PCs and conventional PCs have similar therapeutic functions and that platelets within components have comparable functional, morphologic and proteomic characteristics [7].
(c)
Fig. 1 The effects of photochemical pathogen reduction treatment (PCT) on IL-27 (a), sOX40L (b) and sCD40L (c) release by resting platelets upon storage. PC samples, untreated control (n = 15) and treated with PCT (n = 15) were kept in suspension medium at 22 – 2°C under constant and gentle agitation and rotation for 7 days (a, b, c). Results are presented as means – standard deviations. *P < 0.05 in comparison with day 0 (One-way ANOVA). No analyte/time-point demonstrated a significantly different result between the control and treated units (Kruskal– Wallis Test).
Several groups addressed the issue of the influence of amotosalen of the proteomics of PRT treated vs. untreated platelets [8, 9]. Because some debate persists on the broad implementation of PRT [6] regardless of consensus [10], it appears valuable to carry on investigations on both the physiopathology of intercept treated-platelets and post marketing clinical trials. © 2014 International Society of Blood Transfusion Vox Sanguinis (2015) 108, 205–207
Platelet and cytokines 207
Acknowledgements The authors acknowledge the support of all personnel at the Etablissement Francßais du Sang Auvergne-Loire and the anonymous blood donors. Financial support was
received through grants from the EFS Auvergne-Loire, France, the ANR (ANR-12-JSV1-0012-01), the ANSM (AAP-2012-011), the ART and the Association ‘Les Amis de Remi’. The authors reported no potential conflicts of interest.
References 1 Osselaer JC, Messe N, Hervig T, et al.: A prospective observational cohort safety study of 5106 platelet transfusions with components prepared with photochemical pathogen inactivation treatment. Transfusion 2008; 48:1061–1071 2 Cognasse F, Osselaer JC, Payrat JM, et al.: Release of immune modulation factors from platelet concentrates during storage after photochemical pathogen inactivation treatment. Transfusion 2008; 48:809–813 3 Lin L, Dikeman R, Molini B, et al.: Photochemical treatment of platelet concentrates with amotosalen and long-wavelength ultraviolet light inactivates a broad spectrum of pathogenic bacteria. Transfusion 2004; 44:1496– 1504 4 Hamzeh-Cognasse H, Damien P, Nguyen KA, et al.: Immune-reactive
soluble OX40 ligand, soluble CD40 ligand, and interleukin-27 are simultaneously oversecreted in platelet components associated with acute transfusion reactions. Transfusion 2014; 54:613–625 5 Lin L, Cook DN, Wiesehahn GP, et al.: Photochemical inactivation of viruses and bacteria in platelet concentrates by use of a novel psoralen and longwavelength ultraviolet light. Transfusion 1997; 37:423–435 6 Hervig T, Seghatchian J, Apelseth TO: Current debate on pathogen inactivation of platelet concentrates–to use or not to use? Transfus Apher Sci 2010; 43:411–414 7 Vetlesen A, Mirlashari MR, Akkok CA, et al.: Biological response modifiers in photochemically pathogen-reduced versus untreated apheresis platelet
© 2014 International Society of Blood Transfusion Vox Sanguinis (2015) 108, 205–207
concentrates. Transfusion 2013; 53:147–155 8 Hechler B, Ohlmann P, Chafey P, et al.: Preserved functional and biochemical characteristics of platelet components prepared with amotosalen and ultraviolet A for pathogen inactivation. Transfusion 2013; 53:1187– 1200 9 Jackman RP, Muench MO, Heitman JW, et al.: Immune modulation and lack of alloimmunization following transfusion with pathogen-reduced platelets in mice. Transfusion 2013; 53:2697–2709 10 Cook RJ, Heddle NM: Clinical trials evaluating pathogen-reduced platelet products: methodologic issues and recommendations. Transfusion 2013; 53:1843–1855
Copyright of Vox Sanguinis is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use.
SHORT REPORT
Amotosalen-HCl-UVA pathogen reduction does not alter poststorage metabolism of soluble CD40 ligand, Ox40 ligand and interkeukin-27, the cytokines that generally associate with serious adverse events H. Hamzeh-Cognasse,1 S. Laradi,1,2 J. C. Osselaer,3 F. Cognasse1,2 & O. Garraud1,4 1
Universite de Lyon, GIMAP-EA3064, Saint Etienne, France EFS Auvergne-Loire, Saint Etienne, France 3 Service re gional vaudois de transfusion sanguine, Epalinges, Switzerland 4 Institut National de Transfusion Sanguine (INTS), Paris, France 2
Received: 6 July 2014, revised 28 August 2014, accepted 5 September 2014, published online 21 October 2014
Platelets in therapeutic platelet concentrates are commonly acknowledged to release biologically active constituents during storage. This study examined the influence of photochemical pathogen reduction treatment (PRT) using amotosalen-HCl and UVA light vs. untreated control platelet components, on three factors recently reported to be associated with serious adverse events associated with platelet component (PC) transfusions: sCD40L, IL-27 and sOX40 ligand. Levels of such cytokine-like factors increased significantly during storage, but no significant difference was detected between PRT- and control PCs. This suggests that occurrences of AEs are not directly influenced by PRT but rather may depend on alternate determinants. Key words: cytokine, inflammation, pathogen reduction treatment, platelet storage.
Introduction Pathogen reduction technologies (PRTs) are applicable to platelet components (PCs) with the aim of providing three main benefits: eradication of bacterial contaminations, reduction/inactivation of most known and emergent viruses and inactivation of residual leucocytes. Over a near 10-year experience with this process in some blood establishments (BEs), the Intercept system (amotosalen/HCl/UVA illumination) proved successful on all three targets [1]. PRTs have not been implemented universally yet, mainly because of cost impact, loss of platelets in the PCs and residual concerns on platelet function alteration. It remains relevant to examine whether there is a variation in the platelet capacity for secretion of cytokines and other biological response modifiers (BRMs) that are characteristic of Correspondence: Olivier Garraud, INTS and GIMAP-EA 3064, Universite de Saint-Etienne, Faculte de Medecine, 15 rue Ambroise Pare, 42023 Saint-Etienne Cedex 2, France E-mail: [email protected]
platelet physiology and also present in PCs associated with grade 2–3 serious adverse events, and principally those that are inflammatory in nature [2]. It is worth noticing that reduction of inflammation, suggested as a potential benefit by manufacturers, is not the primary goal of PRTs [3]. Because of novel evidence that certain cytokine-like BRMs are specifically associated with AEs related to PC transfusions, namely sOx40L and IL-27 (along with sCD40L) [4], we aimed at re-evaluating the potential of amotosalenUVA treatment to impact (increase/decrease vs. steady state conditions) BRMs during preparation and storage of PCs, with shelf-life extended to 7 days.
Materials and methods Blood collection using platelet apheresis and PRT were performed (n = 15) as reported previously [5]. Quality control parameters were as follows: mean volume (419 – 30 ml); mean platelet count (5.6 – 0.6 9 1011 platelets/component); mean residual leucocyte count (0.095 – 0.087 9 106 leucocytes/component); and mean 205
206 H. Hamzeh-Cognasse et al.
pH (7.3 – 0.1; d1). All PCs were automatically resuspended in nominal 35% donor plasma and 65% platelet additive solution (InterSol, Fenwal-Europe, Belgium) and divided into paired units (n = 15) of equal volume and platelet content. One component was retained as an untreated control, and the other treated with PRT with 150 lmol/l amotosalen, 3 J/cm2 UVA treatment and CAD absorption). Platelet concentration (106/ml), pH and levels of sCD40L, IL-27 and sOX40L were measured on Days 0–7. Endotoxin levels, sCD40L, IL-27 and sOX40L were measured as described previously and adjusted to 109 platelets [4]. Differences between cytokine levels over time were evaluated by one-way analysis of variance (ANOVA) followed by Bonferroni’s correction. As values were not normally distributed, Interexperiment comparisons of marker expression and cytokine and/or chemokine concentrations were analysed with the nonparametric analysis of variance with the Kruskal–Wallis test using STATVIEW (SAS Institute, Cary, NC, USA) and P values of 0.05) from untreated PCs during 7 days of storage. From Day 1 to 7, the pH of PRTand control PCs decreased similarly, from 7.1 – 0.1 in both D1, to 6.9 – 0.1 and 6.8 – 0.1, respectively, on D7 (data not shown), but remained within acceptable ranges for therapeutic transfusion. IL-27 secretion by platelets in untreated PCs was similar to the sCD40L secretion pattern overtime as previously observed (Fig. 1c), [4] and with a significant increase by day 3 (28600 – 1872 pg/ml *P < 0.05) and a twofold amount of this cytokine on days 5 and 7 (37921 – 2071 and 41234 – 1004 pg/ml *P < 0.05) (Fig. 1a). Soluble (s)OX40L secretion was objectively stable in untreated PCs until day 5 but increased when PCs were stored for another 2 days (d7: 864 – 27 pg/ml, compared to d0: 382 – 12 pg/ml *P < 0.05; Fig. 1b). No differences were detected, however, between PRT- and control PCs regarding all three BRMs, at all times of storage. However, there was a strong trend, concerning elevated IL-27 secretion by platelets in PRT- vs. control PCs, from day 2 of storage (d2, P = 0.066; d3, P = 0.11; d4, P = 0.12; d5, P = 0.075). Several investigators reported that PRT variably affect the levels of platelet-derived cytokines in PCs [6]. Recent investigations, however, show that PRT-PCs and conventional PCs have similar therapeutic functions and that platelets within components have comparable functional, morphologic and proteomic characteristics [7].
(c)
Fig. 1 The effects of photochemical pathogen reduction treatment (PCT) on IL-27 (a), sOX40L (b) and sCD40L (c) release by resting platelets upon storage. PC samples, untreated control (n = 15) and treated with PCT (n = 15) were kept in suspension medium at 22 – 2°C under constant and gentle agitation and rotation for 7 days (a, b, c). Results are presented as means – standard deviations. *P < 0.05 in comparison with day 0 (One-way ANOVA). No analyte/time-point demonstrated a significantly different result between the control and treated units (Kruskal– Wallis Test).
Several groups addressed the issue of the influence of amotosalen of the proteomics of PRT treated vs. untreated platelets [8, 9]. Because some debate persists on the broad implementation of PRT [6] regardless of consensus [10], it appears valuable to carry on investigations on both the physiopathology of intercept treated-platelets and post marketing clinical trials. © 2014 International Society of Blood Transfusion Vox Sanguinis (2015) 108, 205–207
Platelet and cytokines 207
Acknowledgements The authors acknowledge the support of all personnel at the Etablissement Francßais du Sang Auvergne-Loire and the anonymous blood donors. Financial support was
received through grants from the EFS Auvergne-Loire, France, the ANR (ANR-12-JSV1-0012-01), the ANSM (AAP-2012-011), the ART and the Association ‘Les Amis de Remi’. The authors reported no potential conflicts of interest.
References 1 Osselaer JC, Messe N, Hervig T, et al.: A prospective observational cohort safety study of 5106 platelet transfusions with components prepared with photochemical pathogen inactivation treatment. Transfusion 2008; 48:1061–1071 2 Cognasse F, Osselaer JC, Payrat JM, et al.: Release of immune modulation factors from platelet concentrates during storage after photochemical pathogen inactivation treatment. Transfusion 2008; 48:809–813 3 Lin L, Dikeman R, Molini B, et al.: Photochemical treatment of platelet concentrates with amotosalen and long-wavelength ultraviolet light inactivates a broad spectrum of pathogenic bacteria. Transfusion 2004; 44:1496– 1504 4 Hamzeh-Cognasse H, Damien P, Nguyen KA, et al.: Immune-reactive
soluble OX40 ligand, soluble CD40 ligand, and interleukin-27 are simultaneously oversecreted in platelet components associated with acute transfusion reactions. Transfusion 2014; 54:613–625 5 Lin L, Cook DN, Wiesehahn GP, et al.: Photochemical inactivation of viruses and bacteria in platelet concentrates by use of a novel psoralen and longwavelength ultraviolet light. Transfusion 1997; 37:423–435 6 Hervig T, Seghatchian J, Apelseth TO: Current debate on pathogen inactivation of platelet concentrates–to use or not to use? Transfus Apher Sci 2010; 43:411–414 7 Vetlesen A, Mirlashari MR, Akkok CA, et al.: Biological response modifiers in photochemically pathogen-reduced versus untreated apheresis platelet
© 2014 International Society of Blood Transfusion Vox Sanguinis (2015) 108, 205–207
concentrates. Transfusion 2013; 53:147–155 8 Hechler B, Ohlmann P, Chafey P, et al.: Preserved functional and biochemical characteristics of platelet components prepared with amotosalen and ultraviolet A for pathogen inactivation. Transfusion 2013; 53:1187– 1200 9 Jackman RP, Muench MO, Heitman JW, et al.: Immune modulation and lack of alloimmunization following transfusion with pathogen-reduced platelets in mice. Transfusion 2013; 53:2697–2709 10 Cook RJ, Heddle NM: Clinical trials evaluating pathogen-reduced platelet products: methodologic issues and recommendations. Transfusion 2013; 53:1843–1855
Copyright of Vox Sanguinis is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use.
