Convalescent plasma, an apheresis research project targeting and motivating the fully recovered COVID 19 patients: A rousing message of clinical benefit to both donors and recipients alike

This concise manuscript aims to make suggestions for a small step forward in both preventative and therapeutic measures against the Coronavirus disease 2019 (COVID 19) pandemic. This targeted strategy consists of using fully recovered COVID 19 Heroes, that is, brave volunteers, as the source of antibodies in plasma collected by plasmapheresis or affinity column- derived antibodies, both are sterilised and pathogen inactivated for substitution therapy for use in those populations in need of antibody. This include for use in critically ill COVID 19 patients and as a preventative measure, in those at potential risk of infection as no vaccine is yet available. This would be a small step forward, while we are waiting to produce an effective, validated vaccine and witnessing increasing demands for testing and self-isolation which are the two most effective current strategies. In line with this concept, some methodological aspects of the use of the UVC sterilization of FFP/ cryoprecipitate-depleted FFP or immunoglobulins containing neutralizing antibodies for clinical use against COVID-19 are highlighted. The plasmapheresis procedure is, of course, particularly targeted to male donors, who consist of about 75 % of the COVID-19 population and who are able to undergo multiple double, or even triple plasmapheresis procedures. Moreover, as some of these donors have already been in an induced-hypercoagulable state and prone to thrombosis and DVT, this strategy will be partially aimed at improving their health with the use of citrate based anticoagulants and removal of high molecular weight viscous components which contribute to the untoward clinical effects of DVT. Repeated targeted plasmapheresis or plasma exchange of selected COVID-19 positive individuals would undoubtedly lower their state of hypercoagulability and normalize their hypercoagulability. The recipients of such a derived FFP-product would benefit from the two to 3 doses of viral inactivated antibodies, which could neutralize the viral antigens even at very low concentration if present in the early stage. So, this practice would be a double-edged sword with benefits for both donors and recipients

Effects of Blood Products on Inflammatory Response in Endothelial Cells In Vitro

BACKGROUND: Transfusing blood products may induce inflammatory reactions within the vascular compartment potentially leading to a systemic inflammatory response. Experiments were designed to assess the inflammatory potential of different blood products in an endothelial cell-based in vitro model and to compare baseline levels of potentially activating substances in transfusion products. METHODS: The inflammatory response from pre-activated (endotoxin-stimulated) and non-activated endothelial cells as well as neutrophil endothelial transmigration in response to packed red blood cells (PRBC), platelet concentrates (PC) and fresh frozen plasma (FFP) was determined. Baseline inflammatory mediator and lipid concentrations in blood products were evaluated. RESULTS: Following incubation with all blood products, an increased inflammatory mediator release from endothelial cells was observed. Platelet concentrates, and to a lesser extent also FFP, caused the most pronounced response, which was accentuated in already pre-stimulated endothelial cells. Inflammatory response of endothelial cells as well as blood product-induced migration of neutrophils through the endothelium was in good agreement with the lipid content of the according blood product. CONCLUSION: Within the group of different blood transfusion products both PC and FFP have a high inflammatory potential with regard to activation of endothelial cells. Inflammation upon blood product exposure is strongly accentuated when endothelial cells are pre-injured. High lipid contents in the respective blood products goes along with an accentuated inflammatory reaction from endothelial cells

Pathogen reduction/inactivation of products for the treatment of bleeding disorders:what are the processes and what should we say to patients?

Patients with blood disorders (including leukaemia, platelet function disorders and coagulation factor deficiencies) or acute bleeding receive blood-derived products, such as red blood cells, platelet concentrates and plasma-derived products. Although the risk of pathogen contamination of blood products has fallen considerably over the past three decades, contamination is still a topic of concern. In order to counsel patients and obtain informed consent before transfusion, physicians are required to keep up to date with current knowledge on residual risk of pathogen transmission and methods of pathogen removal/inactivation. Here, we describe pathogens relevant to transfusion of blood products and discuss contemporary pathogen removal/inactivation procedures, as well as the potential risks associated with these products: the risk of contamination by infectious agents varies according to blood product/region, and there is a fine line between adequate inactivation and functional impairment of the product. The cost implications of implementing pathogen inactivation technology are also considered

Insights into red blood cell storage lesion: Toward a new appreciation

Red blood cell storage lesion (RSL) is a multifaceted biological phenomenon. It refers to deterioration in RBC quality that is characterized by lethal and sub-lethal, reversible and irreversible defects. RSL is influenced by prestorage variables and it might be associated with variable clinical outcomes. Optimal biopreservation conditions are expected to offer maximum levels of RBC survival and acceptable functionality and bioreactivity in-bag and in vivo; consequently, full appraisal of RSL requires understanding of how RSL changes interact with each other and with the recipient. Recent technological innovation in MS-based omics, imaging, cytometry, small particle and systems biology has offered better understanding of RSL contributing factors and effects. A number of elegant in vivo and in vitro studies have paved the way for the identification of quality control biomarkers useful to predict RSL profile and posttransfusion performance. Moreover, screening tools for the early detection of good or poor “storers” and donors have been developed. In the light of new perspectives, storage time is not the touchstone to rule on the quality of a packed RBC unit. At least by a biochemical standpoint, the metabolic aging pattern during storage may not correspond to the currently fresh/old distinction of stored RBCs. Finally, although each unit of RBCs is probably unique, a metabolic signature of RSL across storage variables might exist. Moving forward from traditional hematologic measures to integrated information on structure, composition, biochemistry and interactions collected in bag and in vivo will allow identification of points for intervention in a transfusion meaningful context. © 2016 Elsevier Lt

An Overview of Current Position on Cell Therapy in Transfusion Science and Medicine: From Fictional Promises to Factual and Perspectives from Red Cell Substitution to Stem Cell Therapy

Stem cell therapy is a relatively novel field of investigation, in which either differentiated cells or stem cells capable of differentiation are transplanted into an individual with the objective of yielding specific cell types in the damaged tissue and consequently restoring its function. The most successful example of cell therapy is hematopoietic stem cell transplantation, leading to regeneration of a patient's blood cells, now a widely established procedure for many oncologic and non–oncologic diseases. Innovative cell-based therapies are being developed to replace, regenerate or repair injured, absent, or diseased tissues and organs. However, cell therapy bioproducts are based on their inherent biological features such as proliferation, migratory, capability, plasticity, and capacity of self-renewal, posing serious challenges during such bioproduct development. The extraordinary promise of stem cells for future treatments of otherwise intractable diseases has raised great hope and expectations in patients, advocates, physicians, and researchers alike. However, despite thousands of scientific publications and research programs, increased efforts need to be put into the identification of the factors involved, biological mechanisms and materials that affect safety/ efficacy, and into the design of cost-effective methods for the harvesting, expansion, manipulation and purification of the cells

Trends and targets of various types of stem cell derived transfusable RBC substitution therapy: Obstacles that need to be converted to opportunity

A shortage of blood during the pandemic outbreak of COVID-19 is a typical example in which the maintenance of a safe and adequate blood supply becomes difficult and highly demanding. So far, human RBCs have been produced in vitro using diverse sources: hematopoietic stem cells (SCs), embryonic SCs and induced pluripotent SCs. The existing, even safest core of conventional cellular bioproducts destined for transfusion have some shortcoming in respects to: donor –dependency variability in terms of hematological /immunological and process/ storage period issues. SCs–derived transfusable RBC bioproducts, as one blood group type for all, were highly complex to work out. Moreover, the strategies for their successful production are often dependent upon the right selection of starting source materials and the composition and the stability of the right expansion media and the strict compliance to GMP regulatory processes. In this mini-review we highlight some model studies, which showed that the efficiency and the functionality of RBCs that could be produced by the various types of SCs, in relation to the in-vitro culture procedures are such that they may, potentially, be used at an industrial level. However, all cultured products do not have an unlimited life due to the critical metabolic pathways or the metabolites produced. New bioreactors are needed to remove these shortcomings and the development of a new mouse model is required. Modern clinical trials based on the employment of regenerative medicine approaches in combination with novel large-scale bioengineering tools, could overcome the current obstacles in artificial RBC substitution, possibly allowing an efficient RBC industrial production

Update on extracellular vesicles inside red blood cell storage units: Adjust the sails closer to the new wind

Release of vesicles from cells is a universal biological system, an adaptive cellular response to endogenous or external physiological or stressful stimuli and a genius means for intercellular, inter-organ and even inter-organism communication. These secreted vesicles that are collectively designated extracellular vesicles (EVs) have increasingly attracted the interest of cell biologists due to their imaginable interactions with every piece of the known biological systems in both health and disease states. Although EVs isolation and characterization are challenges, owing to their particular physicochemical features and complex biology, recent technological innovation has offered better understanding and inevitably, driven the revision of previously established theories on them. However, a crucial question remains unsolved: the physiological relevance of EVs in vivo. Since membrane vesiculation is an integral part of red blood cell (RBC) aging and homeostatic machinery and a prominent feature of RBC storage lesion, the characterization of storage EVs and their probable clinical relevance with the therapeutic or adverse effects of transfusions are extremely important targets in the research fields of transfusion biology and medicine. The scientists involved should transfer nascent knowledge and state-of-the-art technological tools in the packed RBC unit in order to: (i) update the inventory of biochemical and biophysical features of storage EVs; (ii) gain insight into the molecular pathways/signals underlying their generation; and (iii) clarify their dependence on blood donor, storage strategies and analytical variations, in order to step forward on understanding their interactions with stored or recipient target cells. © 2016 Elsevier Lt

Reflection on passive immunotherapy in those who need most: some novel strategic arguments for obtaining safer therapeutic plasma or autologous antibodies from recovered COVID -19 infected patients

The COVID-19 pandemic is an emerging new human disease, for which no vaccines, or monoclonal antibodies (mAbs) or drugs, are currently available for therapy. Active vaccination requires the induction of an immune response against a given agent in a susceptible individual for the pur- pose of preventing or treating an infectious disease and this usually takes time to develop. Thus, the use of existing autol- ogous Ab administration, obtainable from recovered COVID-19 patients two weeks after recovery, is the best and the most practical strategy for providing immediate passive immunity to susceptible recipients in need. Recently, the use of convalescent blood-derived products was proposed by one of the authors of this paper (JS) as an early option for treating patients with Ebola virus disease. In this mini-report we propose three potential additional options for sources of such autologous Ab and provide some operational and evidence-based arguments to support the urgent implementation of such strategic approaches to saving the lives of those in need: (i) the use of hyperimmune immunoglobulin concentrates, which are derived from the plasma of physiologically immunized donors. It is debatable that this method may be even more effective than plasma- pheresis since it uses a smaller dose of about 200 ml, which causes higher donor variability compared to the product of plasmapheresis which provides 600 ml that can be used as a triplet of satellite bags for three recipients

NEW HORIZONS ON STEM CELL CRYOPRESERVATION THROUGH THE ARTIFICIAL EYES OF CD 34+, USING MODERN FLOW CYTOMETRY TOOLS

Hematopoietic stem cell (HSC) cryopreservation is a critical step in autologous and cord blood transplantation (CBT). In most circumstances, cryopreservation is performed in a mixture containing dimethyl sulfoxide (DMSO), since DMSO is necessary to secure cell viability. Most centers use a controlled rate (slow) freezing before the long-term storage at vapor phase liquid nitrogen (LN2) temperatures (≤ −160 °C). The primary objectives for laboratories supporting HSCT programs are to provide secure storage for leukapheresis and cord blood products, and to adequately characterize the functional properties of the grafts before their infusion. In the autologous setting, the large majority of the published results dealt with the assessment of the graft before cryopreservation. On the contrary, in CBT, before a CB unit is released, a sample obtained from a contiguous segment of that CB unit needs to be tested to verify HLA type and cell viability. The effects of graft handling, cryopreservation, storage and thawing on the recovery of CD34+ cells needs to be carefully analyzed and standardized on a global level. Some technical unresolved issues still limit the application of the ISHAGE derived single platform flow cytometry protocol for the assessment of the thawed material; based on these considerations, an adaptation of both the acquisition setting and the gating strategyis necessary for reliable measurement of CD34-expressing HSC in cryopreserved grafts. Artificial intelligence applied to “big data” may provide a new tool for improving advanced processing procedures and quality management guidelines in this area of investigation