Scotblood 2011 features advances in translational medicine, haemopoietic progenitor cells and milestones of blood banking systems.

Amongst the presentations featured at Scotblood 2011 were advances in diagnostic tests for antibodies to red blood cells, the establishment of an islet isolation laboratory, and the development of a clinical product for corneal stem cell transplantation. In addition, the conference comprised presentations on state-of-the-art in collection, storage, and clinical utility of haemopoietic progenitor cells. It also included a session on blood banking systems dedicated to an SNBTS colleague, the late Russell Graham. Finally, the keynote lecture was delivered by Prof. John Forsythe on behalf of the Safety of Blood, Tissues and Organs (SaBTO) members, while the Iain Cook Memorial Lecture was delivered by the recently retired SNBTS R&D director, Prof. Chris Prowse.

Scotblood 2010: key presentations of the past, present, and future of transfusion medicine to mark Scottish national blood transfusion service (SNBTS) anniversaries.

The year 2010 marked the 80th anniversary of the first volunteer blood donor panel in Scotland and the 70th anniversary of the first meeting of the Scottish National Blood Transfusion Association - the forerunner of today's SNBTS. As such the annual Scotblood meeting hosted a distinguished group of speakers to present key note and award lectures on all aspects of Transfusion Medicine including red cell antigens, solving the problems, hazards that shaped our practice, the transfusion needs of patients, donor issues, and component therapy to cellular therapy and beyond. The Iain Cook Memorial Lecture was given by Prof. Dame Marcela Contreras and was entitled "Blood Transfusion International - A Partnership with the Developing World".

The cellular immunobiology associated with fetal and neonatal alloimmune thrombocytopenia.

Fetal and neonatal alloimmune thrombocytopenia (FNAIT) is caused by maternal antibodies that cross the placenta in connection with pregnancy and destroy fetal platelets. Recently, maternal T cell responses associated with FNAIT have been studied at the clonal level. These T cell clones recognize an integrin ß3 epitope, which is anchored to the HLA-DRB3∗0101-encoded MHC molecule DR52a. The same MHC allele is strongly associated with FNAIT. As the production of pathological antibodies reactive with fetal platelets is likely dependent on these T cell responses, there exists a potential for preventing FNAIT by targeting these T cells.

SCOTBLOOD 2009: the quest for understanding vCJD; Claudia's Trachea implantation; transfusion triggers; Scottish histocompatibility and immunogenetics network; and islet cell transplantation.

Scotblood 2009 consisted of a varied combination of leading edge presentations incorporating the past, present and future. Variant CJD was a major feature of the meeting comprising the quest for its understanding and the impact the disease was having on blood donation. The meeting also included the fascinating and groundbreaking story of Claudia's Trachea transplantation, along with progress in the establishment of the Scottish histocompatibility and immunogenetics network and islet transplantation. The meeting began however with a talk on facilities for the future, outlining major modernization of SNBTS in order to meet future challenges.

The relationship of anti-HPA-1a amount to severity of neonatal alloimmune thrombocytopenia - Where does it stand?

The issue of whether or not antibody quantity during pregnancy is related to severity of neonatal alloimmune thrombocytopenia remains unresolved. In this article we cite studies in support of both sides of the argument and highlight some of the reasons that may lie behind the observed differences amongst those studies. It may well be that some of the reasons for the discrepant results could be due to the type of study carried out (eg retrospective versus prospective), the sample size, the timing of antibody sampling, and possibly the type or protocol of assay used. Another major reason is the absence, until recently, of an international anti-HPA-1a standard.

Direct comparison between two quantitative assays in the measurement of maternal anti-HPA-1a antibody in neonatal alloimmune thrombocytopenia (NAIT).

BACKGROUND: Around 80% of severe neonatal alloimmune thrombocytopenia (NAIT) cases in the Caucasian population are due to anti-HPA-1a antibodies. However, the relationship between anti-HPA-1a antibody quantity and severity of NAIT has been subject to debate, possibly due to discrepant results between studies incorporating different assays (such as ELISA and MAIPA) and the number of samples. The aim of this study was to compare the level of maternal anti-HPA-1a antibodies in the same samples, using two different methods; quantitative ELISA and quantitative MAIPA. At the same time, the relationship between the maternal anti-HPA-1a antibody quantity and the newborn platelet count was examined. MATERIALS AND METHODS: Plasma samples were obtained from HPA-1a negative mothers giving birth to children with different platelet counts (i.e. NAIT vs normal platelet count). The antibody levels were quantified blindly in the respective assays using standards calibrated against the international standard NIBSC 03/152. RESULTS: A linear correlation was observed between quantitative MAIPA and quantitative ELISA results. However, there were many individual variations giving a mean ratio of disagreement between the quantitative ELISA and quantitative MAIPA of 2.84. Furthermore, regression analysis revealed a significant relation between anti-HPA-1a antibody quantity measured by MAIPA, but not ELISA, and the newborn platelet count. CONCLUSION: The results indicate that differences observed between various studies with regard to NAIT severity and anti-HPA-1a quantity could partly be due to different assays used.

Scotblood 2008 celebrates 60th anniversary of the NHS through past, present, and future of transfusion medicine, encompassing Hepatitis C Scottish Odyssey, nursing, translational medicine and trauma.

On the occasion of the 60th anniversary of the UK national health service (NHS), Scotblood 2008 featured an opulent array of presentations encompassing the beginnings of the scottish national blood transfusion service (SNBTS), through featuring progress on hepatitis C, to the advancing role of nursing in transfusion medicine, translational medicine, and trauma encountered in military transfusion. This commentary comprises summaries of the presentations, based in part on the abstracts provided by the speakers.

Scotblood 2007: Tackling local and global issues in transfusion medicine - donor recruitment, effective use of blood, stem cell plasticity, and vCJD.

This commentary briefly highlights some of the local and the global contemporary issues affecting transfusion medicine worldwide. The main areas of focus addressed this year were: donor recruitment, stem cell plasticity, the effective use of blood, and vCJD.

What's happening - Scotblood 2006 advances in immunetolerance, information technology, microarrays and pathogen inactivation in transfusion medicine.

This commentary on Scotblood meeting aimed to provide a highlight on four areas of new development in blood transfusion medicine, based on the abstracts provided by the invited speakers. Dr. Jerard Seghatchian would like to express his sincere thanks to Prof. Robin Fraser, Chairman of the Scotblood Organising Committee, and Dr. Hagop Bessos, chairman of the Scotblood Programme Sub-Committee, for inviting him to attend this event and providing him with the essential material for this report.

Prospective epidemiologic study of the outcome and cost-effectiveness of antenatal screening to detect neonatal alloimmune thrombocytopenia due to anti-HPA-1a.

BACKGROUND: To assess the value of antenatal screening to detect neonatal alloimmune thrombocytopenia (NAIT) due to anti-HPA-1a, a prospective study was carried out to quantify the potential clinical benefits and determine whether screening would be cost-effective. STUDY DESIGN AND METHODS: An observational prospective controlled study was carried out on 26,506 pregnant women over 2 years. HPA-1a phenotyping was performed in the first trimester and women confirmed HPA-1a-negative were tested for anti-HPA-1a during pregnancy, at delivery, and 10 to 14 days after birth. Babies of HPA-1a-negative women were tested at delivery for thrombocytopenia and examined for signs of bleeding. Economic evaluation was undertaken on the basis of the data collected during the study. RESULTS: Twenty-five of 318 women (7.9%) had anti-HPA-1a detected for the first time. Eight women (43 per 100,000) gave birth to babies with NAIT, and 5 (27 per 100,000) had severe thrombocytopenia. Three babies had mild signs of bleeding, and no cases of intracranial hemorrhage (ICH) or fetal loss were detected. It is estimated that it would cost 60,596 pounds (98,771 US dollars) to detect a case of severe NAIT, where anti-HPA-1a has been identified for the first time, and 1,151,323 pounds (1,876,656 US dollars) to prevent a case of ICH, assuming that detection allowed successful intervention. CONCLUSIONS: Our data suggest that severe HPA-1a NAIT is underdiagnosed in the absence of routine antenatal screening. Serious bleeding complications and ICH, however, occur less frequently in first cases of NAIT than suspected from the literature, and the costs of screening and possible intervention must be balanced against the procedural risks.

What's happening? The expanding role of apheresis platelet support in neonatal alloimmune thrombocytopenia: current status and future trends.

Despite some unresolved problems that may be associated with platelet transfusion, such as alloimmunisation, refractoriness, bacterial contamination, and the potential side effects related to the development of some biological response modifiers during storage, platelet therapy remains the most effective treatment for the management and prevention of severe thrombocytopenia and hemorrhage [Seghatchian J, Snyder EL, Krailadsiri P, editors. Platelet therapy: current status and future trends. Amsterdam, The Netherlands: Elsevier; 2000]. This appears to be particularly the case in neonatal alloimmune thrombocytopenia (NAIT), which arises due to an incompatibility of the platelet specific antigens between the pregnant mother and her baby. Over 80% of severe NAIT cases in the Caucasian population occur when the mother and baby differ in their HPA-1 epitope (HPA-1a negative and positive respectively) leading, in 10% of cases, to the production of anti-HPA-1a which can cross the placenta and cause NAIT in utero or post-partum. Anti-HPA-5b is the second most cause of NAIT, although severe cases occur only after the first pregnancy. Clinical manifestations of NAIT vary from mild (petechia and bruises) to severe (intracranial haemorrhage with possible death or life long morbidity). A recent study in Scotland indicated that the cost per case of severe NAIT detected during screening of pregnant women, where anti-HPA-1a is detected for the first time, would amount to $98,771 [Turner M, Bessos H, et al. Prospective epidemiological study of the outcome and cost effectiveness of antenatal screening to detect neonatal alloimmune thrombocytopenia (NAIT) due to anti-HPA-1a. Transfusion, in press. Yet, unlike the case with Rhesus hemolytic disease of the new born, the cost-effectiveness of HPA-1 screening in NAIT remains unresolved, as does the most optimal mode of treatment. Therefore, in the absence of a consensus on the screening and optimal management of NAIT, the availability and provision of HPA-1a/5b negative apheresis platelets based on current practice (transfusionguidelines.org.uk) appear to be a clinically effective treatment in NAIT. In that vein, an increasing number of blood transfusion centres are screening blood donors in order to secure panels of donors for the prompt provision of HPA-1a/5b negative apheresis platelets. However, evidence is also accumulating that while platelets derived from various apheresis technologies currently in use may be equivalent in terms of cellular contents (thus meeting specifications), they may differ in terms of the platelet storage lesion, microvesiculation and the development of platelet-derived cytokines and some other biological response modifiers [Seghatchian J. Platelet storage lesion: the influence of various leukoreduction procedures on generation/retention of some biological response modifiers, microvesiculation, distribution of membrane-bound/soluble Prion and the rate of HLA-CLASS1 release. Trans Apher Sci, in press. This manuscript summarises strategy and progress both in the improvement of apheresis platelet quality and provision in NAIT.

The application of a new quantitative assay for the monitoring of integrin-associated protein CD47 on red blood cells during storage and comparison with the expression of CD47 and phosphatidylserine with flow cytometry.

BACKGROUND: After the introduction of universal leukoreduction, the role of factors other than white blood cells in red cell (RBC) storage lesion is attracting increasing attention. These include changes in the levels of CD47 and phosphatidylserine (PS) markers on RBCs during storage. The aim of this study was to monitor these changes with both flow cytometry (FACS) and a newly developed quantitative enzyme-linked immunosorbent assay (ELISA). STUDY DESIGN AND METHODS: A new quantitative ELISA (monoclonal antibody immobilization of RBC antigens [MAIRA]) was developed. The assay yielded consistent linear curves that enabled the measurement of CD47 expression on RBCs. In addition, FACS was used to measure both CD47 expression and PS on RBCs (n = 3 units) during storage (Days 4, 10, 24, and 31). RESULTS: A significant reduction in CD47 expression was observed both by MAIRA assay and by FACS by Days 24 and 31 (p < 0.01), and the correlation between the two assays was significant (p < 0.01). In addition, a significant increase in PS was observed by the same storage days with FACS (p < 0.01). CONCLUSION: The MAIRA assay appears to be suitable for the quantitative measurement of RBC markers during storage. Significant changes in CD47 and PS levels were observed during storage, which may have detrimental immunomodulatory and hemostatic effects on the transfused RBCs.

Characterization of the alloreactive helper T-cell response to the platelet membrane glycoprotein IIIa (integrin-beta3) in human platelet antigen-1a alloimmunized human platelet antigen-1b1b women.

BACKGROUND: The aims were to characterize the helper T-cell response to platelet (PLT) glycoprotein (GP) IIIa, which stimulates the alloimmune antibody response to human PLT antigen (HPA)-1a, to identify immunodominant epitopes and to examine the HLA Class II associations. STUDY DESIGN AND METHODS: Peripheral blood mononuclear cells (PBMNCs) were obtained from 21 HPA-1b1b women who had an HPA-1a-mismatched pregnancy, 14 of whom developed anti-HPA-1a, and 11 control donors. PBMNCs were stimulated with two panels of 15-mer peptides corresponding to the HPA-1a/1b polymorphic region, with either Leu33 (-1a) or Pro33 (-1b) at each possible position, and the proliferative responses were measured. HLA Class II and HPA genotyping was by conventional polymerase chain reaction-sequence-specific priming. RESULTS: Peptides with Leu33 at, or near, the C-terminus contained an immunodominant epitope, stimulating proliferation by helper T cells from all nine women who had anti-HPA-1a at the time of testing; peptide L1 (Val19-Leu33) stimulated a response in 50 percent of these women. Their T cells did not respond to the corresponding HPA-1b Pro33 peptides, and responses to either peptide panel were rare in unimmunized women and controls. HLA-DRB3*01+ was significantly overrepresented (p = 0.014) in alloimmunized women whose T cells responded to the major HPA-1a Leu33-containing epitope. Conversely, HLA-DRB1*15 was negatively associated (p = 0.014) with this response. CONCLUSIONS: The HPA-1a polymorphic region of GPIIIa contains both the linear T-cell and the conformational B-cell epitopes. The immunodominant T-cell epitope is constrained by HLA-DRB3*01+, and if presented by a tolerogenic route, a peptide containing this epitope may form the basis for the prevention or reversal of the alloimmune response to HPA-1a.

Red cell storage lesion: the potential impact of storage-induced CD47 decline on immunomodulation and the survival of leucofiltered red cells.

Red blood cells undergo major biochemical and biomechanical changes during storage that could effect their post transfusion performance. Biochemical effects include changes in 2,3-diphosphoglycerate (2,3-DPG), ATP, and calcium levels, as well as metabolic modulation and release of Annexin V, a cytosolic component of blood cells, as a global marker of cellular injury and fragmentation. Biomechanical changes include alterations in cellular membrane, shape changes, phospholipid content, phospholipid asymmetry, and antigenic markers. Although the extent of these changes under various storage conditions has been well documented, their clinical effects remain unclear. In the current era of universal leucodepletion, the immunomodulatory effects of some essential markers such as CD47 and phosphatidyl serine become the focus of interest as highlighted in this manuscript.

The development of a quantitative ELISA for antibodies against human platelet antigen type 1a.

BACKGROUND: Severe neonatal alloimmune thrombocytopenia is often due to antibodies against human platelet antigen type 1a (HPA-1a). The aim of this study was to develop a quantitative ELISA for the measurement of antibodies against HPA-1a. STUDY DESIGN AND METHODS: HPA-1a glycoprotein (GP) IIb-IIIa was immobilized and mixed with recalcified anti-HPA-1a-positive plasma overnight at 4 degrees C. The beads were washed, the antibodies against HPA-1a were eluted, and the eluate pH level was promptly adjusted. The purified antibodies were dialyzed and used for the development of an ELISA incorporating HPA-1a-coated plates. RESULTS: Serial doubling dilutions of the purified antibodies resulted in consistent sigmoid standard curves with a sensitivity of 0.5 microg per mL. To determine the reproducibility of the ELISA, antibodies against HPA-1a in five plasma samples (Samples A-E) were measured at serial doubling dilutions in four separate assays. Three of the samples (Samples A-C) contained antibodies against HPA-1a. The mean amounts in microg per mL (+/- SD, percentage of CV) obtained in the four assays were as follows: Sample A, 133 (9.4, 7.1%); Sample B, 16.5 (1.7, 10%); and Sample C, 8 (0.8, 10%). The amounts in the two antibody-negative controls (Samples D and E) were consistently less than 0.2 microg per mL. CONCLUSION: Using immobilized HPA-1a1a, antibodies against HPA-1a has been purified, and a quick and simple quantitative assay has been developed.