Mark Scott

By Mark D. Scott, Wendy M. Toyofuku, Xining Yang, Meera Raj and Ning Kang

While ABO/Rh(D) red blood cells (RBC)-matched transfusions are generally considered as safe, a significant risk of alloimmunization to non-A/B blood group antigens exists; especially in chronically transfused patients. Indeed, alloimmunization to non-A/B antigens can be so severe that RBC transfusion can no longer be safely administered without the risk of a potentially deadly immune haemolytic reaction. Currently, no satisfactory solutions exist either to prevent blood group alloimmunization or to cost-effectively treat patients with severe alloimmunization. To address this problem, we have pioneered the immunocamouflage of donor RBC. The immunocamouflaged (stealth) RBC is manufactured by the covalent grafting of biologically safe polymers to RBC membrane proteins. As a result of the grafted polymer, non-A/B blood group antigens are biophysically and immunologically masked. Of particular interest is the immunocamouflage of the Rh(D) antigen which could be used to improve blood inventory and transfusion safety. The polymer-modified RBCs are morphologically normal and, in mice, exhibit normal in vivo survival at immunoprotective grafting concentration. In this chapter, we explore both the biophysical and immunological consequences of the grafted polymers, explore the conditions in which they might be appropriately used, and describe the technology necessary to manufacture functional transfusable units of these cells within the clinical setting.

Part of the book: Transfusion Medicine and Scientific Developments

By Mark D. Scott, Duncheng Wang, Wendy M. Toyofuku and Xining Yang

T cells are key mediators of graft tolerance/rejection, development of autoimmunity, and the anticancer response. Consequently, differentially modifying the T cell response is a major therapeutic target. Most immunomodulatory approaches have focused on cytotoxic agents, cytokine modulation, monoclonal antibodies, mitogen activation, adoptive cell therapies (including CAR-T cells). However, these approaches do not persistently reorient the systemic immune response thus necessitating continual therapy. Previous murine studies from our laboratory demonstrated that the adoptive transfer of polymer-grafted (PEGylated) allogeneic leukocytes resulted in the induction of a persistent and systemic tolerogenic state. Further analyses demonstrated that miRNA isolated from the secretome of polymer-modified or control allogeneic responses effectively induced either a tolerogenic (TA1 miRNA) or proinflammatory (IA1 miRNA) response both in vitro and in vivo that was both systemic and persistent. In a murine Type 1 diabetes autoimmune model, the tolerogenic TA1 therapeutic effectively attenuated the disease process via the systemic upregulation of regulatory T cells while simultaneously downregulating T effector cells. In contrast, the proinflammatory IA1 therapeutic enhanced the anticancer efficacy of naïve PBMC by increasing inflammatory T cells and decreasing regulatory T cells. The successful development of this secretome miRNA approach may prove useful treating both autoimmune diseases and cancer.

Part of the book: Cells of the Immune System

By Mark D. Scott, Nobu Nakane and Elisabeth Maurer-Spurej

Unlike red blood cells (RBC) which are stored at 4°C, platelets are stored at 22–24°C (room temperature) due to biophysical and biochemical changes induced by cold temperatures aggregately known as the ‘cold storage lesion’ (CSL). However, 22°C storage greatly increases the risk of microbial growth, thus limiting the safe storage of platelets to only 5–7 days (versus 42 days for RBC). Consequent to the short shelf life of platelets, blood services face chronic shortages of these life-saving cells. To overcome both the risk of microbial contamination and the constrained supplies of platelets, renewed research into attenuating the CSL and/or determining where cold stored platelets are clinically suitable are ongoing. In this chapter, we show that the covalent grafting of methoxypolyethylene glycol (mPEG), a biocompatible polymer, to the membrane of platelets attenuates the CSL. Moreover, the grafted mPEG serves as a potent cryoprotectant allowing platelets to be stored at 4°C, or frozen at −20°C, while retaining normal platelet counts and biologic function. The successful development of platelet PEGylation may provide a means by which the cold storage of platelets can be achieved with a minimal loss of platelet quality while improving both platelet microbial safety and inventory.

Part of the book: Cryopreservation

By Mark D. Scott, Kerryn Matthews and Hongshen Ma

Among the most crucial rheological characteristics of blood cells within the vasculature is their ability to undergo the shape change (i.e., deform). The significance of cellular deformability is readily apparent based solely on the disparate mean size of human erythrocytes (~8 μm) and leukocytes (10–25 μm) compared to the minimum luminal size of capillaries (4–5 μm) and splenic interendothelial clefts (0.5–1.0 μm) they must transit. Changes in the deformability of either cell will result in their premature mechanical clearance as well as an enhanced possibility of intravascular lysis. In this chapter, we will demonstrate how microfluidic devices can be used to examine the vascular deformability of erythrocytes and agranular leukocytes. Moreover, we will compare microfluidic assays with previous studies utilizing micropipettes, ektacytometry and micropore cell transit times. As will be discussed, microfluidics-based devices offer a low-cost, high throughput alternative to these previous, and now rather ancient, technologies.

Part of the book: Current and Future Aspects of Nanomedicine

By Mark D. Scott

β thalassemias arise from genetic defects that interfere with the synthesis of the β hemoglobin chain and the subsequent production of the normal α2β2 hemoglobin tetramer. As a consequence of this decreased β-chain synthesis, unpaired α-hemoglobin chains are found within the red blood cell (RBC). The unstable α-chains are associated with a number of cellular defects, including: membrane-bound globin; membrane thiol oxidation; altered cytoskeletal proteins; decreased cellular and membrane deformability; and increased susceptibility to both endogenous and exogenous oxidants. Surprisingly, while significant injury to human thalassemic RBC arise from the unpaired α-chains, the underlying intra-RBC mechanisms are not easily studied in patient samples or in mouse models. To better study the fate of excess α-chains in human RBC, the model β Thalassemic cell was developed. Model human β thalassemic RBC is made by entrapping purified human α-chains within normal RBC via osmotic lysis and resealing. This human model allows for the systematic examination of the mechanisms underlying the α-chain mediated damage in the β thalassemic RBC. Studies utilizing the model β thalassemic RBC have demonstrated that the α-chains give rise to an iron and glutathione-dependent, self-amplifying and self-propagating oxidative reaction.

Part of the book: Beta Thalassemia

By Xining Yang and Mark D. Scott

T cell-mediated immunomodulation can be, in simple terms, defined as altering the normal Treg:Teff ratio. Immunosuppression skews the net Treg:Teff ratio toward the ‘tolerogenic’ Treg component, while immunoactivation skews the response toward the ‘proinflammatory’ Teff component. In the treatment of autoimmune diseases, achieving an immunosuppressive state is a desirable goal in order to prevent ongoing injury by activated Teff cells. In contrast, an innate, or induced, immunosuppressive state can be deleterious and prevent pathogen-induced disease while allow for the progression of cancer. Indeed, a current goal of cancer therapy is attenuating an existing endogenous immunosuppressive state that prevents effective T cell-mediated immunorecognition of cancer cells. Thus, the biological modulation of the Treg:Teff ratio provides a unique approach for treating both autoimmune diseases and cancers. Using a biomanufacturing system, miRNA-enriched immunotherapeutic has been generated that either induce (TA1) or overcome (IA1) an immunosuppressive state. As will be shown, these therapeutics show efficacy both in vitro and in vivo in the prevention of autoimmune Type 1 diabetes and in enhancing the ability of resting immune cells to recognize and inhibit cancer cell growth. The successful development of these cost-effective, and easily biomanufactured, secretome-based therapeutics may prove useful in treating both autoimmune diseases and cancer.

Part of the book: Immunosuppression