Puget Sound Blood Center Research Institute

Seattle, WA, United States

Puget Sound Blood Center Research Institute

Seattle, WA, United States

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Munday A.D.,Puget Sound Blood Center Research Institute | Lopez J.A.,Puget Sound Blood Center Research Institute
Blood | Year: 2013

In this issue of Blood, Kasahara et al report that platelet-dependent clot retraction requires factor XIII (FXIII), which covalently associates.brin polymers with protein located within the platelet plasma membrane at lipid rafts. © 2013 by The American Society of Hematology.


Zimring J.C.,Puget Sound Blood Center Research Institute | Zimring J.C.,University of Washington
Blood | Year: 2015

The collection and storage of red blood cells (RBCs) is a logistical necessity to provide sufficient blood products. However, RBC storage is an unnatural state, resulting in complicated biological changes, referred to collectively as the "storage lesion." Specifics of the storage lesion have been studied for decades, including alterations to cellular properties, morphology, molecular biology of carbohydrates, proteins and lipids, and basic metabolism. Recently, mass spectrometry-based "omics" technology has been applied to the RBC storage lesion, resulting in many new observations, the initial effects of which are more information than understanding. Meanwhile, clinical research on RBC transfusion is considering both the efficacy and also the potential untoward effects of transfusing stored RBCs of different ages and storage conditions. The myriad biological changes that have now been observed during the storage lesion have been extensively reviewed elsewhere. This article focuses rather on an analysis ofourcurrentunderstanding of the biological effects of different elements of the storage lesion, in the context of evolving new clinical understanding. A synopsis is presented of both established and theoretical considerations of the RBC storage lesion and ongoing efforts to create a safer and more efficacious product. © 2015 by The American Society of Hematology.


Diz-Kucukkaya R.,Nightingale | Lopez J.A.,Puget Sound Blood Center Research Institute | Lopez J.A.,University of Washington
Hematology/Oncology Clinics of North America | Year: 2013

Platelet membrane glycoproteins play a key role in hemostasis and thrombosis. Although disorders of platelet membrane glycoproteins are rare, their effects on the lives of those affected are very important. Severe deficiencies manifest themselves early during childhood with mucocutaneous bleeding. Mild deficiencies may not be diagnosed until adulthood or until the hemostatic system is stressed by surgery or trauma. The diagnosis of these disorders requires detailed laboratory investigation. Management of bleeding in patients with inherited platelet disorders requires both preventive measures and the treatment of individual bleeding episodes according to severity. The study of platelet membrane disorders also has yielded important insights into the functions of affected proteins, information that has produced some of the most successful antithrombotic drugs currently in use. © 2013 Elsevier Inc.


Patel S.R.,Emory University | Zimring J.C.,Puget Sound Blood Center Research Institute | Zimring J.C.,University of Washington
Transfusion Medicine Reviews | Year: 2013

Traditionally, alloimmunization to transfused blood products has focused exclusively on recipient antibodies recognizing donor alloantigens present on the cell surface. Accordingly, the immunologic sequelae of alloimmunization have been antibody mediated effects (ie, hemolytic transfusion reactions, platelet refractoriness, anti-HLA and anti-HNA effects, etc). However, in addition to the above sequelae, there is also a correlation between the number of antecedent transfusions in humans and the rate of bone marrow transplant (BMT) rejection-under reduced intensity conditioning with HLA-matched or HLA-identical marrow. Bone marrow transplant of this nature is the only existing cure for a series of nonmalignant hematologic diseases (eg, sickle cell disease, thalassemias, etc); however, rejection remains a clinical problem. It has been hypothesized that transfusion induces subsequent BMT rejection through immunization. Studies in animal models have observed the same effect and have demonstrated that transfusion-induced BMT rejection can occur in response to alloimmunization. However, unlike traditional antibody responses, sensitization in this case results in cellular immune effects, involving populations such as T cell or natural killer cells. In this case, rejection occurs in the absence of alloantibodies and would not be detected by existing immune-hematologic methods. We review human and animal studies in light of the hypothesis that, for distinct clinical populations, enhanced rejection of BMT may be an unappreciated adverse consequence of transfusion, which current blood bank methodologies are unable to detect. © 2013 Elsevier Inc.


Zimring J.C.,Puget Sound Blood Center Research Institute | Zimring J.C.,University of Washington | Spitalnik S.L.,Columbia University
Annual Review of Pathology: Mechanisms of Disease | Year: 2015

Antibody-induced hemolytic transfusion reactions were first described over 300 years ago. Indeed, during its early evolution, transfusion medicine focused almost exclusively on issues in immunohematology to prevent such events. However, despite the best of efforts to avoid them, incompatible transfusions still occur, through both error and an inability to obtain compatible red blood cells for patients who are alloimmunized against multiple antigens. Because transfusing units of incompatible blood is potentially lethal, studies on human volunteers are not ethical. Thus, understanding of hemolytic transfusion reactions is generated through clinical cases, animal models, inference from related human pathologies, or studies using small volumes of transfused red blood cells. Over the past several decades, substantial new knowledge has been accumulated regarding the mechanisms of hemolysis, the metabolism of products of hemolysis, and the effects of both on recipient biology. Using these data sources, this article traces the historical generation of this knowledge and describes recent advances. © 2015 by Annual Reviews.


Kim Y.C.,Uniformed Services University of the Health Sciences | Zhang A.-H.,Uniformed Services University of the Health Sciences | Su Y.,Uniformed Services University of the Health Sciences | Rieder S.A.,National Institute of Allergy and Infectious Diseases | And 5 more authors.
Blood | Year: 2015

Expansion of human regulatory T cells (Tregs) for clinical applications offers great promise for the treatment of undesirable immune responses in autoimmunity, transplantation, allergy, and antidrug antibody responses, including inhibitor responses in hemophilia A patients. However, polyclonal Tregs are nonspecific and therefore could potentially cause global immunosuppression. To avoid this undesirable outcome, the generation of antigen-specific Tregswould be advantageous. Herein,we report the production and properties of engineered antigen-specific Tregs, created by transduction of a recombinant T-cell receptor obtained from a hemophilia A subject's T-cell clone, into expanded human FoxP3+ Tregs. Such engineered factor VIII (FVIII)-specific Tregs efficiently suppressed the proliferation and cytokine production of FVIII-specific T-effector cells. Moreover, studies with an HLA-transgenic, FVIII-deficient mouse model demonstrated that antibody production from FVIII-primed spleen cells in vitro were profoundly inhibited in the presence of these FVIII-specific Tregs, suggesting potential utility to treat anti-FVIII inhibitory antibody formation in hemophilia A patients. (Blood. 2015;125(7):1107-1115) Copyright 2011 by The American Society of Hematology. All rights reserved.


Johnsen J.,Puget Sound Blood Center Research Institute
Hematology / the Education Program of the American Society of Hematology. American Society of Hematology. Education Program | Year: 2012

Idiopathic (immune) thrombocytopenic purpura (ITP) is a common autoimmune disorder resulting in isolated thrombocytopenia. ITP can present either alone (primary) or in the setting of other conditions (secondary) such as infections or altered immune states. ITP is associated with a loss of tolerance to platelet antigens and a phenotype of accelerated platelet destruction and impaired platelet production. Although the etiology of ITP remains unknown, complex dysregulation of the immune system is observed in ITP patients. Antiplatelet antibodies mediate accelerated clearance from the circulation in large part via the reticuloendothelial (monocytic phagocytic) system. In addition, cellular immunity is perturbed and T-cell and cytokine profiles are significantly shifted toward a type 1 and Th17 proinflammatory immune response. Further clues into immune dysregulation in ITP may be gleaned from studies of secondary ITP. Some infections can induce antiplatelet Abs by molecular mimicry, and there may be common elements involved in breaking tolerance with other autoimmune disorders. There is also evidence for a genetic predisposition to both ITP and responsiveness to therapy, which may in part lie within immune-related genes. Lastly, treatment with immunomodulatory agents remains the mainstay of ITP therapies.


Wallace N.A.,Fred Hutchinson Cancer Research Center | Robinson K.,Fred Hutchinson Cancer Research Center | Howie H.L.,Puget Sound Blood Center Research Institute | Galloway D.A.,Fred Hutchinson Cancer Research Center
PLoS Pathogens | Year: 2015

Recent work has explored a putative role for the E6 protein from some β-human papillomavirus genus (β-HPVs) in the development of non-melanoma skin cancers, specifically β-HPV 5 and 8 E6. Because these viruses are not required for tumor maintenance, they are hypothesized to act as co-factors that enhance the mutagenic capacity of UV-exposure by disrupting the repair of the resulting DNA damage. Supporting this proposal, we have previously demonstrated that UV damage signaling is hindered by β-HPV 5 and 8 E6 resulting in an increase in both thymine dimers and UV-induced double strand breaks (DSBs). Here we show that β-HPV 5 and 8 E6 further disrupt the repair of these DSBs and provide a mechanism for this attenuation. By binding and destabilizing a histone acetyltransferase, p300, β-HPV 5 and 8 E6 reduce the enrichment of the transcription factor at the promoter of two genes critical to the homology dependent repair of DSBs (BRCA1 and BRCA2). The resulting diminished BRCA1/2 transcription not only leads to lower protein levels but also curtails the ability of these proteins to form repair foci at DSBs. Using a GFP-based reporter, we confirm that this reduced foci formation leads to significantly diminished homology dependent repair of DSBs. By deleting the p300 binding domain of β-HPV 8 E6, we demonstrate that the loss of robust repair is dependent on viral-mediated degradation of p300 and confirm this observation using a combination of p300 mutants that are β-HPV 8 E6 destabilization resistant and p300 knock-out cells. In conclusion, this work establishes an expanded ability of β-HPV 5 and 8 E6 to attenuate UV damage repair, thus adding further support to the hypothesis that β-HPV infections play a role in skin cancer development by increasing the oncogenic potential of UV exposure. © 2015 Wallace et al.


Zimring J.C.,Puget Sound Blood Center Research Institute
Transfusion | Year: 2014

Background Red blood cell (RBC) transfusion is a lifesaving therapy, the logistic implementation of which requires RBC storage. However, stored RBCs exhibit substantial donor variability in multiple characteristics, including hemolysis in vitro and RBC recovery in vivo. The basis of donor variability is poorly understood. Study Design and Methods We applied a murine model of RBC storage and transfusion to test the hypothesis that genetically distinct inbred strains of mice would demonstrate strain-specific differences in RBC storage. In vivo recoveries were determined by monitoring transfused RBCs over 24 hours. Timed aliquots of stored RBCs were subjected to tandem chromatography/mass spectrometry analysis to elucidate metabolic changes in the RBCs during storage. Results Using independent inbred mouse strains as donors, we found substantial strain-specific differences in posttransfusion RBC recovery in vivo after standardized refrigerated storage in vitro. Poor posttransfusion RBC recovery correlated with reproducible metabolic variations in the stored RBC units, including increased lipid peroxidation, decreased levels of multiple natural antioxidants, and accumulation of cytidine. Strain-dependent differences were also observed in eicosanoid generation (i.e., prostaglandins and leukotrienes). Conclusion These findings provide the first evidence of strain-specific metabolomic differences after refrigerated storage of murine RBCs. They also provide the first definitive biochemical evidence for strain-specific variation of eicosanoid generation during RBC storage. The molecules described that correlate with RBC storage quality, and their associated biochemical pathways, suggest multiple causal hypotheses that can be tested regarding predicting the quality of RBC units before transfusion and developing methods of improved RBC storage. © 2013 American Association of Blood Banks.


Lopez J.A.,Puget Sound Blood Center Research Institute | Zheng Y.,University of Washington
Journal of Thrombosis and Haemostasis | Year: 2013

The microvasculature is an immense organ that defines the environmental conditions within tissues in both health and disease, and is vital for the proper functions of all tissues. Here, we describe existing tools to study vascular cell function and our work using one platform of in vitro microvessels, which we employed to study vessel structure and remodeling, endothelial barrier function, angiogenesis, interactions between endothelial cells and perivascular cells, interactions between blood cells and the endothelium, and microvascular thrombosis. We also briefly discuss the potential future applications of these platforms in biology and medicine. © 2013 International Society on Thrombosis and Haemostasis.

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