Katherine Serrano

Liposome—complement interactions in rat serum: implications for liposome survival studies

Serum complement opsonizes particles such as bacteria for clearance by the reticuloendothelial system. Complement has been reported to interact with liposomes and therefore may mediate the reticuloendothelial system clearance of liposomes. This study has used a rat serum model to define some of the characteristics of liposomes which modulate their ability to activate complement. Using functional hemolytic assays and C3/C3b crossed immunoelectrophoresis, we have demonstrated that liposomes activated rat complement in a dose-dependent manner with higher concentrations of liposomes activating higher levels of complement. The detection of complement activation required the inclusion of phospholipids bearing a net charge. Complement activation occurred via the classical pathway; no alternative pathway activation was detected. The presence of cholesterol contributed to complement activation in a dose-dependent manner. Phospholipid fatty acyl chain length did not influence complement activation while the introduction of unsaturated acyl chains markedly decreased levels of complement activation. Liposome size also influenced complement activation with 400 nm unilamellar vesicles more effectively activating complement than 50 nm vesicles for equivalent amounts of exposed lipid. These studies demonstrate that the composition of the liposome greatly affects the in vitro activation of rat serum complement and suggest that the biological half-life of liposomes in the circulation of rats may be altered by changing the liposome composition to reduce complement activation.

The Platelet Storage Lesion

The gradual loss of quality in stored platelets as measured collectively with various metabolic, functional, and morphologic in vitro assays is known as the platelet storage lesion. With the advent of pathogen reduction technologies and improved testing that can greatly reduce the risk for bacterial contamination, the platelet storage lesion is emerging as the main challenge to increasing the shelf life of platelet concentrates. This article discusses the contribution of platelet production methods to the storage lesion, long-established and newly developed methods used to determine platelet quality, and the significance for clinical transfusion outcome. Highlighted are the novel technologies applied to platelet storage including platelet additive solutions and pathogen inactivation.

Comprehensive proteomic analysis of protein changes during platelet storage requires complementary proteomic approaches.

BACKGROUND: Proteomics methods may be used to analyze changes occurring in stored blood products. These data sets can identify processes leading to storage‐associated losses of blood component quality such as the platelet (PLT) storage lesion (PSL). The optimal strategy to perform such analyses to obtain the most informative data sets, including which proteomics methods, is undefined. This study addresses relative differences among proteomics approaches to the analysis of the PLT storage lesion.

A comparative study of common techniques used to measure haemolysis in stored red cell concentrates

Background and Objectives  There is no standardized method of measuring the parameters for haemolysis determination of red cell concentrate (RCC). Three haemoglobin quantification methods (automated analyser, Harboe and Drabkin’s) and two methods of haematocrit measurement (automated analyser and microcapillary centrifugation) were evaluated for use with RCC.

Effects of prestorage white cell reduction on platelet aggregate formation and the activation state of platelets and plasma enzyme systems.

BACKGROUND: The introduction of prestorage white cell (WBC) reduction in random‐donor platelet concentrates in Canada has increased the occurrence of particulate material in PCs. The effects of filtration on platelet activation state and the activation of plasma enzyme systems were assessed.

Adjuvant treatment for complement activation increases the effectiveness of photodynamic therapy of solid tumors

Phototoxic lesions generated in tumor tissue by photodynamic therapy (PDT) are recognized by the host as a threat to the integrity and homeostasis at the affected site. Among the canonical pathways invoked by the host for dealing with this type of challenge is the activation of the complement system, integrating proteins that serve as molecular sensors of danger signals produced by PDT and those initiating signalling cascades coupled into the network of inflammatory and immune responses. Since the activated complement system is a salient participant of the antitumor response produced by PDT, it is worth exploring whether its manipulation can be exploited for the therapeutic benefit. Using mouse tumor models, the present study examined the potential of representative complement-activating agents to act as effective adjuvants to PDT. Tumor-localized treatment with zymosan, an alternative complement pathway activator, reduced the recurrence-rate of PDT-treated tumors, markedly increasing the percentage of permanent cures. In contrast, a similar treatment with heat aggregated gamma globulin (complement activator via the classical pathway) was of no significant benefit as a PDT adjuvant. Systemic complement activation with streptokinase treatment had no detectable effect on complement deposition at the tumor site without PDT, but it augmented the extent of complement activity in PDT-treated tumors. This finding based on immunohistochemistry analysis explains the results of tumor therapy experiments, which showed that systemic treatment with streptokinase or a similar agent, urokinase, enhances the PDT-mediated tumor response. Zymosan and streptokinase administrations produced no beneficial results with PDT of tumors growing in complement-deficient mice. This study, therefore, establishes the potential of complement-activating agents to serve as effective adjuvants to PDT for cancer treatment.

Plasma and cryoprecipitate manufactured from whole blood held overnight at room temperature meet quality standards.

BACKGROUND: With buffy coat (BC) processing of whole blood (WB) donations, the preparation of plasma occurs within 24 hours rather than 8 hours of collection. The effect of this change on coagulation factor function in plasma and cryoprecipitate was evaluated during the validation of this production method and with routine production.

Whole blood treated with riboflavin and ultraviolet light: quality assessment of all blood components produced by the buffy coat method.

Pathogen inactivation (PI) technologies are currently licensed for use with platelet (PLT) and plasma components. Treatment of whole blood (WB) would be of benefit to the blood banking community by saving time and costs compared to individual component treatment. However, no paired, pool‐and‐split study directly assessing the impact of WB PI on the subsequently produced components has yet been reported.

The effect of timing of gamma-irradiation on hemolysis and potassium release in leukoreduced red cell concentrates stored in SAGM

While irradiation of red cell concentrates (RCC) prevents graft‐versus‐host disease in susceptible transfusion recipients, it also damages red blood cells (RBC). To understand the ability of irradiation regulations to prevent transfusion of inferior units, we irradiated 980 RCC in saline‐adenine‐glucose‐mannitol (SAGM) using various combinations of pre‐irradiation age and post‐irradiation storage times, and measured hemolysis and extracellular potassium levels. We observed unacceptably high hemolysis (>0·8%) in some RCC and elevated extracellular potassium levels in all gamma‐irradiated RCC. This suggests that more restrictive storage times should be considered for RCC in SAGM.

Development of a quality monitoring program for platelet components: a report of the first four years' experience at Canadian Blood Services

BACKGROUND: A quality monitoring program (QMP) for platelet concentrates (PCs) was implemented at Canadian Blood Services (CBS) to improve standards and to better understand platelet (PLT) products by supplementing routine quality control (QC).