Cryopreservation alters the membrane and cytoskeletal protein profile of platelet microparticles

Raynel, Sarah; Padula, Matthew P.; Marks, Denese C.; Johnson, Lacey

doi:10.1111/trf.13165

Cited by 53 publications

(85 citation statements)

References 51 publications

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“…Previous in-vitro studies suggested that this was due to significant changes in cryo-PLT properties and increased soluble factors within the PC resulting in altered platelet functionality such as haemostasis and anticoagulation [172,177,[180][181][182][183][184]. Given the changes in the cryo-PLT product [172,177,[180][181][182][183][184], and the capacity of PC to significantly alter recipients' immune responses [89,162,[166][167][168] The capacity of DC to initiate immunity is dependent upon their maturation stage, accompanied by phenotypic changes associated with antigen presentation [193,195].…”

Section: Discussionmentioning

confidence: 99%

“…The process of platelet cryopreservation for transfusion has been reported to modify cellular function [172,[180][181][182][183][184]. Valeri and colleagues (2005) However, in comparison to conventional PC, in-vitro studies have reported that cryo-PLT are different in a number of parameters.…”

Section: "Cryo-plt Storage Lesion": Biochemical and Biomechanical Modmentioning

confidence: 99%

“…These modifications have been associated with a significantly higher number of nondiscoid platelets [181], as well as microparticles expressing apoptosis mediators, such as PS, and other cytoskeletal membrane proteins within the unit itself [180,183,184]. Studies have reported that the cryo-PLT unit contains significantly higher numbers of PS-and tissue factor-positive microparticles than conventional PC [180,183].…”

Section: "Cryo-plt Storage Lesion": Biochemical and Biomechanical Modmentioning

confidence: 99%

“…During the process of platelet cryopreservation (cooling to very low temperatures, typically -80 o C) and preparation for transfusion (thawing at 37°C) platelet properties and function may be modified [172,[180][181][182][183][184]. In-vitro studies have reported significant reduction in cryo-PLT recovery [177], aggregation [172,180,181] and hypotonic stress response [182] in comparison to conventional PC.…”

Section: Introductionmentioning

confidence: 99%

“…These modifications are associated with a significantly higher proportion of non-discoid platelets [181] as well as microparticles expressing apoptotic mediators including PS, and other cytoskeletal membrane proteins (e.g. including actin, filamin, gelsolin, and tropomyosin) within the unit [180,183,184]. These changes may be due to the process of cryopreservation inducing cryoinjury due to excessive cellular dehydration, osmotic injury and extracellular ice crystal formation [191].…”

Section: Introductionmentioning

confidence: 99%

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Characterisation of dendritic cell immune profile in models of transfusion

Ki¹

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Blood transfusion modulates recipients' immune responses sometimes resulting in poor clinical outcomes. Further, transfusion with blood products at date-of-expiry has been associated with exacerbation of this concerning problem. In-vitro studies have demonstrated exposure to packed red blood cell (PRBC) and platelet products modulate the responses of leukocytes, however, there are limited studies on the impact of transfusion on dendritic cells (DC). Despite DC being central to the immune response, their role in transfusion-related immune modulation remains largely undefined. I hypothesised that exposure to blood components changes DC phenotype and function, which could be one mechanism underpinning transfusion-related immune modulation. The potential for blood components (PRBC, buffy-coat-derived platelet concentrates (PC) and cryopreserved platelets (cryo-PLT)) to modulate responses of myeloid dendritic cells ii whether blood components exposing F-actin for Clec9A ligation were present within the blood product. rhClec9A did not bind fresh or stored PRBC, and this outcome was supported by the lack of detection of F-actin. Although, no binding of rhClec9A to PC or cryo-PLT was demonstrated, F-actin was detected on both types of platelet products. As binding of rhClec9A to PRBC, PC or cryo-PLT was not detected, EDTA blood collection tubes were used for the remainder of the study under the premise that lack of expression of this receptor would not impact on outcomes in my transfusion models.I assessed the impact of fresh and/or stored blood products on mDC and BDCA3 + DC surface antigen and inflammatory profile using a human in-vitro whole blood model of transfusion. In parallel, to model the processes associated with viral or bacterial infection, polyinosinic:polycytidylic acid (polyI:C) or lipopolysaccharide (LPS) was added respectively. Exposure to PRBC and PC predominately suppressed surface antigen expression (CD40, CD80, CD83 and CD86) and inflammatory mediator production (interleukin (IL)-6, IL-8, IL-12, tumour necrosis factor-α and interferon-gamma inducible protein-10 or IL-10) on both DC subsets. These changes were often more evident in the presence of polyI:C and LPS, and when DC were exposed to stored PRBC. Similar modulation was evident for BDCA3 + DC when exposed to cryo-PLT alone and cryo-PLT in the presence of polyI:C and LPS. The impact of blood transfusion on the overall inflammatory response by leukocytes was also examined. The immunomodulatory effect of transfusion in-vitro was more pronounced in the presence of polyI:C and LPS. For PRBC, an additional erythrophagocytosis assay was conducted where the uptake of stored PRBC by mDC and BDCA3 + DC was significantly increased in comparison to fresher PRBC.This study provided the first evidence that exposure of PRBC, PC and cryo-PLT modulates mDC and/or BDCA3 + DC maturation and activation. The changes were more pronounced when modelling processes associated with concurrent viral or bacterial infection.Experimental evidence suggested that mod...

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Section: Discussionmentioning

confidence: 99%

Section: "Cryo-plt Storage Lesion": Biochemical and Biomechanical Modmentioning

confidence: 99%

Section: "Cryo-plt Storage Lesion": Biochemical and Biomechanical Modmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Characterisation of dendritic cell immune profile in models of transfusion

Ki¹

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Cryopreserved Mesenchymal Stromal Cells Are Susceptible to T-Cell Mediated Apoptosis Which Is Partly Rescued by IFNγ Licensing

et al. 2016

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We have previously demonstrated that cryopreservation and thawing lead to altered MSC functionalities. Here, we further analyzed MSC's fitness post freeze-thaw. We have observed that thawed MSC can suppress T-cell proliferation when separated from them by transwell membrane and the effect is lost in a MSC:T-cell coculture system. Unlike actively growing MSCs, thawed MSCs were lysed upon coculture with activated autologous PBMCs and the lysing effect was further enhanced with allogeneic PBMCs. The use of DMSO-free cryoprotectants or substitution of HSA with human platelet lysate in freezing media and use of autophagy or caspase inhibitors did not prevent thaw defects. We tested the hypothesis that IFNγ pre-licensing before cryobanking can enhance MSC fitness post thaw. Post thawing, IFNγ licensed MSCs inhibit T cell proliferation as well as fresh MSCs and this effect can be blocked by 1-methyl Tryptophan, an IDO inhibitor. In addition, IFNγ prelicensed thawed MSCs inhibit the degranulation of cytotoxic T cells while IFNγ unlicensed thawed MSCs failed to do so. However, IFNγ prelicensed thawed MSCs do not deploy lung tropism in vivo following intravenous injection as well as fresh MSCs suggesting that IFNγ prelicensing does not fully rescue thaw-induced lung homing defect. We identified reversible and irreversible cryoinjury mechanisms that result in susceptibility to host T-cell cytolysis and affect MSC's cell survival and tissue distribution. The susceptibility of MSC to negative effects of cryopreservation and the potential to mitigate the effects with IFNγ prelicensing may inform strategies to enhance the therapeutic efficacy of MSC in clinical use.

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