Search

Objective To analyze the serological characteristics and clinical significance of IgG anti-D and anti-C combined antibodies and anti-E and anti-c combined antibodies in patients negative for RhDC and RhEc antigens. Methods The clinical data and laboratory results of 12 cases with two types of irregular antibodies were recorded and analyzed, including age, sex, history of blood transfusion/pregnancy, ABO and RhD blood group identification, Rh antigen typing, irregular antibody screening, antibody-specific identification, absorption-elution tests, antibody titer determination and cross-matching tests. Results Among the 12 patients, the mean age was 51.4±16.9 years. Nine patients had a history of blood transfusion; eight patients had a history of pregnancy; five patients had both. Serological tests showed positive antibody screening and incompatible cross-matching. The results of antibody-specific identification and absorption-elution tests showed the presence of both IgG anti-D and anti-C antibodies in three patients, with anti-D titers at 16-32, and anti-C titers at 8-16. Nine patients had both IgG anti-E and anti-c antibodies, with the titers of anti-E and anti-c antibodies at 8-16. From the patients with combined anti-D and anti-C antibodies, suspended red blood cells of ABO identical type, RhD negative and other Rh antigens as ccee were selected. From patients with combined anti-E and anti-c antibodies, suspended red blood cells of ABO identical type, RhD positive and other Rh antigens as CCee were selected. Cross-matching blood test results showed no agglutination or hemolysis in saline, polycoagulant and anti-human globulin media. Conclusion Blood transfusion and/or pregnancy are the primary causes of irregular antibodies in two Rh systems, leading to positive antibody screening and cross-match incompatibility. Routine compatibility transfusion of Rh antigens, based on ABO homotypic blood transfusion, is of great value and significance for the safety of clinical blood transfusion and the prevention of hemolytic disease of the newborn.

Maternal alloantibodies directed against fetal red blood cell (RBC) antigens may cause potentially life-threatening haemolytic disease of the fetus and newborn (HDFN). Dutch transfusion guidelines therefore prescribe preventive cEK matching for all (pre-)fertile females. To quantify the impact of cEK matching, we compared overall and antigen-specific cumulative RBC alloimmunisation incidences in females and males aged <45 years. Among a multicentre cohort comprised of patients who received their first and subsequent RBC unit between 2005 and 2019, first-formed RBC alloantibodies were detected in 47 of 2998 (1·6%) females and 49 of 2507 (2·0%) males. Comparing females and males, overall alloimmunisation incidences were comparable (3·1% [95% confidence interval (CI) 2·1-4·4] versus 3·5% (95% CI 2·4-4·9, P = 0·853) after 10 units transfused). However, cEK alloimmunisation incidences were significantly lower among females (0·6% (95% CI 0·3-1.5) versus 2·2% (95% CI 1·5-3·4, P = 0·001) after 10 units transfused). Yet, despite cEK-matching guidelines being in effect, 6·5%, 3·6% and 0·2% of all RBC units remained mismatched for c, E or K antigens respectively. Most of these mismatches were almost always due to emergency settings. Even though cEK alloimmunisation was not prevented completely, implementation of cEK matching resulted in an alloantigen-exposure risk reduction of up to 98%., (© 2021 The Authors. British Journal of Haematology published by British Society for Haematology and John Wiley & Sons Ltd.)

Red blood cell alloimmunisation after transfusion of red blood cell concentrates carries a risk for every recipient. This risk is particularly high for patients with conditions such as sickle cell disease. However, red blood cell alloimmunisation can also occur after platelet concentrate transfusion. All blood group systems other than ABO are affected, and there are several mechanisms responsible for this alloimmunisation. The practical implications of this are a need to match red blood cell concentrates in all alloimmunised patients and, in pregnant women, recongnition of the risk of developing haemolytic disease of the foetus and newborn. Several measures can be taken to prevent alloimmunisation: in the case of the D antigen, for example, anti-RhD immunoglobulins can be infused before transfusing platelet concentrates from an RhD-positive donor in a RhD-negative recipient., (Copyright © 2020 Société française de transfusion sanguine (SFTS). Published by Elsevier Masson SAS. All rights reserved.)

Although the electronic cross-matching of blood has been widely applied in some developed countries and regions, concern over the risk of undetected red blood cell (RBC) antibodies has limited its application in mainland China.This study was performed to explore the missed detection rate of RBC antibodies in a Chinese population from 2011 to 2016. If the results of 2 consecutive tests of ABO/RhD blood group identification were consistent and antibody screening results were negative, electronic cross-matching of the blood was performed. In addition, traditional serological cross-matching of blood (polybrene method) and a parallel experiment for electronic cross-matching of blood were performed to analyze the missed detection of unexpected RBC antibodies in blood donors and recipients.Using the polybrene method, 40,228 blood samples were tested by parallel traditional serological cross-matching of blood; among these samples, blood compatibility was found in 40,222 cases, primary incompatibility (incompatibility of the donor's erythrocytes with the recipient's serum) was found in 6 cases, and no secondary incompatibility was found. Identification of antibody specificity was performed using panel cells, and all unexpected RBC antibodies were confirmed as anti-Mur alloantibodies in the MNS system.Further improvements in the erythrocyte antigenic spectrum, especially the Mur antigen in Asian populations, are expected to ensure the safety of implementing electronic cross-matching in China.

Background: Passenger lymphocyte syndrome occurs when donor lymphocytes are transplanted with a solid organ and produce alloantibodies that react with antigens on the recipient's red blood cells (RBCs). Typically, passenger lymphocyte syndrome presents as immunoglobulin G antibody-mediated, extravascular hemolytic anemia with reticulocytosis. Often, the donor was alloimmunized before transplantation., Case Report: A 34-year-old Group O, D+ man with a negative antibody screen received a liver transplant from a Group O, D- donor. Twenty Group O, D+ RBC units were transfused on Postoperative Days (PODs) 0 through 2. On POD 7, the patient developed anemia, a weakly positive antibody screen, and a positive direct antiglobulin test with anti-D in the eluate. After POD 8, a D- transfusion protocol was initiated. Despite laboratory evidence of hemolysis, two initial peripheral blood smears showed no increase in schistocytes or spherocytes, the reticulocyte count was depressed, and a marrow biopsy revealed erythroid hyperplasia. Eventually, anemia resolved after a period of medication non-compliance; however, a positive direct antiglobulin test persisted to the last follow-up date (POD 233)., Results: Other potential causes of aplastic anemia were investigated, but no alternative cause was found. History excluded passive anti-D. D+, LW- cells were reactive, excluding anti-LW. Genotyping showed no evidence of a partial D genotype. Chart review revealed that the liver donor had a history of anti-D. A diagnosis of passenger lymphocyte syndrome was reached., Conclusion: Although antibody-mediated hemolytic anemia has been reported to cause reticulocytopenia in the presence of marrow erythroid hyperplasia, this report of passenger lymphocyte syndrome causing a similar post-transplant anemia in association with reticulocytopenia is noteworthy., (© 2017 AABB.)

Background: Many hospital transfusion services prioritize ABO plasma compatibility in platelet (PLT) transfusion to minimize risk for acute hemolytic transfusion reactions. In spite of the low risk of D alloimmunization associated with apheresis PLT transfusion, attempts may also be made to provide D- PLTs to D- patients. This study was undertaken to assess how often ABO compatibility and/or D matching occurs at our institution and how the ABO and D mix of our PLT supply impacts PLT selection., Study Design and Methods: We retrospectively reviewed the ABO and D type of all PLTs transfused plus the age, sex, and ABO and D type of all PLT recipients between January 2010 and March 2014 (51 months)., Results: We provided ABO-identical PLTs for 5281 (54.6%), ABO plasma-compatible and cellular-incompatible for 3136 (32.4%), ABO low-titer plasma-incompatible and cellular-compatible for 1150 (11.9%), ABO plasma-incompatible and cellular-compatible for 30 (0.3%), and ABO plasma-incompatible and cellular-incompatible for 72 (0.7%). PLT supply did not match PLT demand based on patient ABO type, primarily due to a lower than expected supply of group O PLTs and higher than expected supply of group A and AB. D- patients were less likely to receive ABO-identical PLT transfusions (p = 0.0008), but were more likely to receive D- PLT transfusions (p < 0.0001)., Conclusion: At our hospital, available inventory and PLT selection practices resulted in the majority of group O patients receiving cellular-incompatible PLT transfusions. Efforts to provide D- PLTs to D- patients also resulted in fewer D- patients receiving ABO-identical PLT transfusions., (© 2015 AABB.)

The increasing demand for solid organs has necessitated the use of ABO and Rhesus (Rh) D minor mismatched transplants. The passenger lymphocyte syndrome (PLS) occurs when donor lymphocytes produce antibodies that react with host red blood cell (RBC) antigens and result in hemolysis. Our aim was to evaluate prospectively the role of PLS in post transplant anemia and hemolysis in ABO and RhD minor mismatched recipients of liver and kidney grafts and to study the association of PLS with donor lymphocyte microchimerism. We examined 11 liver and 10 kidney recipients at Day +15 for anemia, markers of hemolysis, direct antiglobulin test and eluates, and serum RBC antibodies. Microchimerism was determined in peripheral blood lymphocytes by genotyping of simple sequence length polymorphisms encoding short tandem repeats. Immune hemolytic anemia and anti-recipient RBC antibodies were observed in 2 out of 11 liver (18.2%) and 2 out of 10 kidney (20%) transplants. RBC antibody specificity reflected the donor to recipient transplant, with anti-blood group B antibodies identified in 2 cases of O to B and 1 case of A to AB transplants while anti-D antibodies were detected in 1 case of RhD-negative to RhD-positive transplant. Donor microchimerism was found in only 1 patient. In conclusion, passenger lymphocyte mediated hemolysis is frequent in minor mismatched liver and kidney transplantation. Recognizing PLS as a potential cause of post transplant anemia may allow for early diagnosis and management to decrease the morbidity and mortality in some patients., (Copyright © 2015 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved.)

Background: In Pakistan routine blood group typing of thalassemia patients identifies ABO and Rh(D) antigens only. Therefore, other antigen incompatibilities between blood donor and blood recipient may cause alloimmunisation., Objective: The aim of this study was to estimate the frequency of alloimmunisation and to evaluate the risk factors associated with its development in beta (β)-thalassemia patients receiving regular blood transfusions., Materials and Methods: In total 162 β thalassemia patients were included in this study. An extended red cell antigen panel was performed to detect antibodies. Patients received red cell concentrates, which were matched for ABO and Rh(D) antigens. Clinical and laboratory data were collected and analysed to estimate the frequency of alloantibodies and the factors influencing immunisation in patients on regular blood transfusion., Results: The median age of patients was 6·7 (range: 0·5-25) years. A total of 14 (8·6%) patients developed alloantibodies against red cell antigens. The most frequently occurring alloantibodies was anti-E (2·5%), anti-K (1·8%), anti-e (1·2%) and anti-D (0·6%). Five (3·1%) patients developed more than one red blood cell (RBC) alloantibody. Age at first transfusion in alloimmunised patients was 1·22 ± 0·87 years. The frequency of blood transfusion in alloimmunised patients was 23 ± 8·81 days and in those without alloimmunisation was 31·8 ± 16 days (p = 0·02). Logistic regression analysis showed no independent risk factor associated with alloimmunisation., Conclusion: The frequency of transfusion was increased in patients who developed alloantibodies. Typing patients and donors to match for Rh and Kell antigens would prevent more than 90% of RBC alloantibodies and reduce the frequency of transfusion in thalassemia patients., (© 2015 British Blood Transfusion Society.)

Background: AABB Standard 5.27 requires transfusion services to have a process for urgent release of blood before completion of compatibility testing. Our institution endorses a policy for the emergency release of group O, D+ red blood cells (RBC; O+ RBC) to males and females at least 50 years of age. Our emergency department (ED) stocks 4 O- RBC units. To determine if O+ RBCs can replace ED O- RBCs, we performed a retrospective review., Study Design and Methods: Patients admitted to the ED between January 2001 and August 2011 and transfused emergency-release O- RBCs were identified. Data were collected on sex, age, length of stay, clinical status, ABO/Rh, RBC transfusions, and RBC antibody screen results., Results: A total of 498 ED O- RBC units were transfused to 268 patients (168 male, 100 female). A total of 322 units were transfused to males and 114 to females at least 50 years of age. Thirty-nine (14%) were D- with 18 receiving O+ RBCs. A total of 109 had follow-up antibody screens; one D- patient developed alloanti-D., Conclusions: The findings support the placement of O+ RBCs in the ED. The majority of ED O- RBCs (88%) went to patients who qualified for O+ RBCs; a minority (1.5%) of patients were D- females less than 50 years of age. The rate of alloimmunization was low., (© 2014 AABB.)

A 5-year-old male with sickle cell disease presented with pain, dark urine, and fatigue 10 days after a red blood cell (RBC) transfusion. Laboratory evaluation demonstrated severe anemia, blood type O+, and anti-D in the serum. Anti-D in a D+ patient led to RH genotyping, which revealed homozygosity for RHD*DAU4 that encodes partial D antigen. Anti-D in this patient whose RBCs exclusively express partial D caused a delayed hemolytic transfusion reaction after exposure to D+ RBCs. The finding of anti-D in a D+patient should be investigated by molecular methods to help distinguish an alloantibody from an autoantibody.

The reported frequency of D alloimmunization in D- recipients after transfusion of D+ platelets varies. This study was designed to determine the frequency of D alloimmunization, previously reported to be an average of 5 ± 2%. A primary anti-D immune response was defined as the detection of anti-D ≥ 28 d following the first D+ platelet transfusion. Data were collected on 485 D- recipients of D+ platelets in 11 centres between 2010 and 2012. Their median age was 60 (range 2-100) years. Diagnoses included: haematological (203/485, 42%), oncological (64/485, 13%) and other diseases (218/485, 45%). Only 7/485 (1·44%; 95% CI 0·58-2·97%) recipients had a primary anti-D response after a median serological follow-up of 77 d (range: 28-2111). There were no statistically significant differences between the primary anti-D formers and the other patients, in terms of gender, age, receipt of immunosuppressive therapy, proportion of patients with haematological/oncological diseases, transfusion of whole blood-derived or apheresis platelets or both, and total number of transfused platelet products. This is the largest study with the longest follow-up of D alloimmunization following D+ platelet transfusion. The low frequency of D alloimmunization should be considered when deciding whether to administer Rh Immune Globulin to D- males and D- females without childbearing potential after transfusion of D+ platelets., (© 2014 John Wiley & Sons Ltd.)

Objective: To determine transfusion compatibility of maternal RBCs for her neonate up to 4 weeks of age, irrespective of maternal-neonatal ABO mismatch., Methods: This was a prospective observational study involving eligible mothers with their neonates delivered in participating site from 1 July 2012 till 31 December 2012. Mother's blood was collected before child birth. Neonatal blood sample was collected from placental end, shortly after birth. Blood Groups of mother-baby pair were individually tested for ABO and Rh-D groups. Pairs with negative Rh-D antigen/s or with same ABO blood groups were excluded. Thus, 28 pairs with both maternal and neonatal samples positive for Rh-D antigen and with different maternal neonatal ABO blood groups were included in the study. Blood samples were collected at birth and at 4 weeks. Cross matching was done at birth and at 4 weeks for each pair with standard blood bank protocols., Results: All 28 pairs showed positive compatibility with standard blood bank cross-matching protocols at birth and at 4 weeks., Conclusions: Maternal blood irrespective of ABO compatibility might be a viable and potentially acceptable option for her new born baby in neonatal period. This may be especially important in developing world with limited blood bank resources.

Background: There is an ongoing controversy regarding provision of usually matched blood (i.e. matched for ABO-D antigens) or phenotypically matched blood (also matched for Rh and Kell antigens) for multiply transfused thalassaemics, especially in developing countries. A pilot study conducted at our centre revealed an alloimmunisation rate of 3.79% with Rh and Kell alloantibodies accounting for 90% of all antibodies. The present cross-sectional study was conducted to assess the impact of a policy of partial better matching (for Rh cDE and Kell antigens) of blood on alloimmunisation in thalassaemics., Material and Methods: In this cross-sectional study three groups of patients were considered. Group 1 comprised 211 thalassaemics who received usually matched (UM) blood until April 2009. Their rates of alloimmunisation have already been published in a prior study. Group 2 consisted of 46 thalassaemics who were enrolled after April 2009 and have received partially better matched (PBM) blood (matched for ABO, Rh cDE and Kell antigens) since the initiation of transfusion therapy. Group 3 (UM→PBM) comprised the patients from group 1 who, from April 2009, were given partial better matched blood. Antibody screening (using a 3-cell panel) and antibody identification (11-cell panel) were carried out to detect the presence of alloantibodies., Results: None of the thalassaemic patients in group 2 (PBM) developed alloantibodies. Eight thalassaemics in group 3 (UM→PBM) developed new alloantibodies (after April 2009)., Discussion: According to the results of the present study, providing at least partially better matched blood appears to improve the efficacy of transfusion for chronically transfused thalassaemics. Large-scale, comprehensive, multicentre studies need to be carried out to formulate realistic, evidence-based, economically feasible transfusion policies for thalassaemic children based on the red blood cell antigen profile of the population.

We describe here a young adult patient with penetrating neck injuries (PNI) with an Rh negative blood type and discuss the perioperative anaesthetic management of single-stage surgical exploration under general anaesthesia and extracorporeal circulation in this patient. The patient had zone II PNI and he was in a haemodynamically progressive unstable state, and the knife penetrated the left internal jugular vein, superior thyroid artery and recurrent laryngeal nerve; the trachea and the oesophagus were swelling at a rapid rate. Eight weeks after operation, the patient was discharged from the hospital without any complications.

The alloimmunization potential of many RHD variants is unknown, and it can be explored by lookback and traceback studies. Hema-Quebec (HQ) investigated the RHD status of 3980 D- repeat blood donors. Thirteen were found to be RHD positive: 4 RHD*1p, and 1 RHD*487delACAG, which show a Dphenotype;and 1 RHD*885T and 7 RHD*(93-94insT) causing a DEL phenotype when C antigen is present. Look back studies were done to verify the alloimmunization potential of these eight DEL donors. Coincidentally, Canadian Blood Services (CBS)performed a trace back study by investigating the RHD status of donors after aD- recipient developed anti-Dafter transfusion of two D- red blood cell (RBC) units. Donor genotyping was done either manually (HQ) or using the Progenika Bloodchip platform(CBS). Donations were traced through computer records. Letters were sent to hospital blood bank physicians to verify the presence of anti-Din recipients and to donors to request repeat samples.A total of 118 RBC units were transfused, 82 to D- recipients.Anti-D was found in three patients transfused with RHD*(93-94insT) DEL red blood cells. One donor presenting the same DEL variant was involved in the trace back study. Even without strong evidence clearly demonstrating the alloimmunization potential of DEL variants, whenever HQ or CBS identifies a donor harboring a DEL phenotype, his or her D status will be changed from DtoD+ to protect against the potential alloimmunization risk.

We report the death of a 61-year-old Japanese man massively transfused during and after emergency aortic surgery. Postoperative on day 8, he died after cardiac arrest associated with hyperkalemia. Indirect antiglobulin testing demonstrated both anti-Di(b) and anti-E antibodies pre-transfusion, and elevation of their titers as the delayed hemolytic transfusion reaction evolved. Monocyte monolayer assay (induction of reactive monocytes) and flow cytometry (increase of IgG1 and/or IgG3) gave evidence of the clinical significance of both antibodies. Anti-Di(b) must be considered when an antibody to a high incidence antigen is found in Japanese and other Mongoloid populations., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Purpose of the Study: To evaluate the prevalence of alloimmunization in women followed in an obstetrical environment in Tunisia, to identify the specificities of antibodies found and to determine factors that could influence the appearance of this immunization., Methods: We proceeded to a retrospective analysis of search for irregular antibodies in women followed up in obstetrical environment over nine consecutive years (2000-2008). The panel was officially defined and produced by the Regional Centre for Blood Transfusion in Sfax (Tunisia)., Results: Overall 5369 women benefited from 6575 antibody testing (average: 1.22; extremes: 1-14). The results were positive for 278 women (5.17 %), allowing to identify 216 antibodies or associations of antibodies. Among identified antibodies, those immune were found in 198 women. The rate of alloimmunization was 3.68 % (198/5369). The majority of the antibodies found was anti-Rh1, isolated or associated with another antibody, in 84.3 % of the total immunized women. The immunization of women according to the number of gestations showed a significant increasing rate ranging from 2.34 % for a first gestation to 5.27 % for four gestations or more. In addition, a significant difference was also noted between the rate of immunization in women who had received anti-Rh1 immunoglobulin and those who had not., Conclusion: Anti-Rh1 immunization is the most frequent in the population of studied women. This could denote of an insufficiency in pregnancies follow-up and immunoprophylaxis protocols., (Copyright © 2012 Elsevier Masson SAS. All rights reserved.)

A 66-year-old man was injured in the traffic accident and showed the signs of hemorrhagic shock. His blood type was Rh negative O. As excessive bleeding was going on, we decided to transfuse 8 units of Rh-incompatible type O red blood cell (RBC) without performing cross match test according to the guideline for treatment of critical bleeding. Though we were aware of the risks of that treatment, no hemolytic reaction nor any other side effect was observed during and after the transfusion. Anti-Rh antibody was not developed either while Anti-Rh immunoglobulin was not given to him. His blood type has been proved to be Del type red blood cell. We conclude that transfusion of Rh-incompatible blood in patient with critical hemorrhage should be encouraged to save life.

Objective: To report a rare case of hemolytic disease of the newborn (HDN) with kernicterus caused by anti-Di(a) diagnosed using high-throughput genotyping multiplex ligation-dependent probe amplification (MLPA)., Methods: Conventional serological methods were used to detect the antibodies related with HDN. The genotypes of more than 40 red blood cell antigens for the newborn and her parents were obtained using the high-throughput MLPA assay. The antibody titers were tested using a standard serological method., Results: The unknown antibody against the low-frequency antigens was predicted based on the primary serological tests. The genotyping results for more than 40 red blood cell antigens of the newborn and her parents showed incompatible antigens of MNS and Diego blood group system, indicating the existence of anti-N or anti-Di(a). Further serological tests confirmed anti-Di(a) existence in the plasma of the newborn and her mother. The titer of anti-Di(a) in the mother's plasma was 1:32., Conclusion: Severe HDN including kernicterus can result from anti-Di(a). High-throughput genotyping MLPA assay can help type some rare antigens in complicated cases. The reagent red cell panels including Di(a)-positive cells are necessary in routine antibody screening test in Chinese population.

Background: Trauma patients receive emergency transfusions of unmatched Type O Rh-negative (Rh-) blood until matched blood is available. We hypothesized that patients given uncrossmatched blood may develop alloantibodies, placing them at risk for hemolytic transfusion reactions (HTRs)., Methods: Data regarding alloantibody profiles and HTR occurrence were collected from the records of trauma patients at our university-based trauma center who received emergency uncrossmatched blood from July 2008 to August 2010., Results: A total of 132 patients received 1,570 units of packed red blood cells. Mean injury severity score was 28 ± 1.3. Forty-five (34%) patients died: 27 on hospital day 1; the remaining 18 had no evidence of HTR before death. Four Rh- female patients received Rh+ fresh frozen plasma, but none received Rh+ packed red blood cells. Three Rh- male patients received both Rh+ packed red blood cells and fresh frozen plasma, and one received Rh+ fresh frozen plasma. One patient developed anti-Rh D antibodies. None experienced HTR. One female patient had HTR from reactivation of anamnestic JK antibodies. Thirteen (33%) of 39 patients met criteria for HTR based on urinalysis and 29 (40%) of 72 patients tested met criteria for HTR based on hemoglobin and bilirubin values. Only one patient had confirmed HTR., Conclusion: High rates of injury recidivism in trauma patients increase the likelihood of multiple blood transfusions during their lifetime. Rh- patients who receive Rh+ blood are at risk of developing anti-Rh antibodies, putting them at risk for HTR. The conservation of Rh- blood for use in female patients may be detrimental to Rh- male patients. Laboratory diagnostic criteria for HTR are nonspecific in the trauma population and should be used with caution.

Transfusion remains the main treatment of sickle cell disease patients. Red cell alloimmunization is frequent because of the antigen disparities between patients of African descent and donors of European ancestry. Alloimmunization is associated with severe hemolytic transfusion reaction, autoantibody formation, and difficulties in the management of transfusion compatibility. Beside common antigens, a number of different RH variant antigens found in individuals of African descent can be involved in alloimmunization. If some variants, such as Hr(S) negative antigens, are known to prone significant alloantibodies and delayed hemolytic transfusion reactions, it is not clear whether all the described variants represent a clinical risk for sickle cell disease patients. The knowledge of the clinical relevance of RH variants is a real issue. An abundance of molecular tools are developed to detect variants, but they do not distinguish those likely to prone immunization from those that are unlikely to prone immunization and delayed hemolytic transfusion reactions. A strategy of prevention, which generally requires rare red blood cells, cannot be implemented without this fundamental information. In this review, we discuss the relevance of RH variants in sickle cell disease, based on the published data and on our experience in transfusion of these patients., (Copyright © 2011 Elsevier Masson SAS. All rights reserved.)

The minor antigen "c" of the rhesus system, which is revealed in population both in rhesus-negative and in rhesus-positive person with 80% frequency, is considered highly immunogenic, it causes severe hemolytic complications while hemotransfusion. In pregnant women it causes the immune conflict and, as a consequence, a hemolytic disease of newborn babies, demanding the treatment, using exchanging blood transfusion (erythrocytes). For the immune conflict prevention it is obligatory to determine the rhesus phenotype in donor and recipient as well as to investigate the presence of alloisoimmune antibodies in a recipient. In hemolytic disease of newborn baby the titer of antibodies towards erythrocytes in a parturient woman blood serum is obligatory estimated with individual matching of erythrocyte-containing media for the exchanging transfusion to newborn baby.

Patient samples were referred to our immunohematology reference laboratory to investigate the presence of a weak D antigen. In the last 3 years, 26 samples were received. Serology and molecular analyses were performed to identify the weak D variant. RHD mRNA from all patients was reverse transcribed, and cDNA was sequenced. The results were compared with a normal RHD sequence to identify the polymorphisms causing the weak D phenotype. Five different already known RHD variants were observed: weak D type 1 (5 individuals), weak D type 2 (1 individual), weak D type 42 (17 individuals), weak D type 45 (1 individual), and partial D DNB (2 individuals). Surprisingly, weak D type 42 was prevalent in our population, whereas weak D type 1, 2, and 3 are the most prevalent variants elsewhere. Anti-D was found in six cases of weak D type 42. The higher prevalence of weak D type 42 could be the result of a founder effect. Additional studies are needed to estimate the frequency of this variant in the general population.

Allogeneic red blood cells transfusion is always an immunological challenge and the choice of the blood products is crucial for the patient safety. But this choice may be hampered by the quality or the quantity of the available supply. In the end, the lack of transfusion may be more harmful than transfusion. The balance between patients' needs and blood centres supplying is always delicate. The conditions are not the same for all blood groups. Things are easier for the KEL1 phenotype, where the supply must ensure only 92.5% of KEL: -1 red blood cells instead of the 91% expected. More complicated is the situation for group O red blood cells with 47 versus 43%. But the major problem concerns RH: -1 red blood cells, for which the needs reach 20.1 versus 15%. These challenges require a lot of efforts from blood centres staffs to influence blood donors' recruitment and appointments. A justified and carefully selected blood products issuing may be of great help, especially for group O RH: -1 red blood cells. Therefore, hospital blood banks must have ad hoc procedures and a trained staff to put them into practice., (Copyright © 2010 Elsevier Masson SAS. All rights reserved.)

Objectives: To evaluate the efficiency of blood tests (blood group, direct antiglobulin test) to assess severe hyperbilirubinemia in full-term newborns, delivered from mothers with rhesus negative or O group and to determine clinical and biological factors that may improve the prediction characteristics of this blood test., Patients and Methods: We included all the full-term newborns, delivered from mothers with rhesus negative or O group, in a tertiary maternity ward, in 2005, from January6th to December31st., Results: One thousand and ninety-two children were included. Newborns of A, B or AB group delivered from a mother 0 were at increased risk of presenting severe hyperbilirubinemia (OR=2.35 [1.22-4.52]). The negative predictive value was 96%. Yet, the determination of the Coombs test does not increase NPV., Conclusion: Systematic performance of blood test for newborns delivered from mother with O group does increase the ability to predict severe hyperbilirubinemia in a newborn infant. Direct antiglobulin test systematic performance remains questionable., (Copyright 2010 Elsevier Masson SAS. All rights reserved.)

HDN attributed to the rare Rh variants has become more and more significant caused by anti-D, but the compatible blood is usually very difficult to obtain when exchange transfusion is required. We treated a 10-hour neonate of O, D + C + c - E - e+ blood group with severe HDN due to anti-Rh17 with least incompatible blood typed O, D + C - c + E + e-. The neonatal hemolysis was relieved obviously and bilirubin was reduced gradually after exchange transfusion. The infant was discharged in good health 13 days after birth with 135.0 g/L, 28.0 micromol/L and 10.7 micromol/L of Hb, total bilirubin and direct bilirubin, respectively. No sequelae were observed in a three-year follow-up. The result suggesting that the least incompatible blood is an alternative choice for exchange transfusion in severe HDN due to anti-Rh17 in case that Rh17 antigen-negative blood is unavailable.

Antibodies of apparent D specificity may be found in D+ patients. We report a D+, multi-transfused Caucasian woman with myelodysplasia who exhibited several alloantibodies. One antibody was a moderately strong (2+) anti-D that persisted for 9 months, until the woman died. Molecular analysis of the patient's RHD gene identified the rare weak D type 21 (938C > T) allele. D alloantibodies do not occur in patients with most weak D types, but some patients with a weak D phenotype, including those with type 21, can produce antibodies to nonself epitopes of the wild-type D antigen.

We investigated the impact of ABO and Rhesus (Rh) blood group matching on the outcome of hematopoietic stem cell transplantation (HSCT) of 154 patients matched at 10/10 HLA loci with unrelated donors. ABO and Rh, as potential risk factors, were modeled with the clinical outcome--acute and chronic graft-versus-host disease (aGVHD, cGVHD), relapse, treatment-related mortality (TRM), and overall survival (OS)--by simple, multiple, and competing risk analyses. We found that minor ABO-mismatches represent a significant risk factor for aGVHD (II-IV) with an estimated risk increase of almost 3-fold (hazard ratio [HR]=2.92, 95% confidence interval [CI]: 1.43-5.95, P=.003), and even 4-fold for aGVHD (III-IV) (HR=4.24, 95% CI: 1.70-10.56, P=.002), but not for other transplant endpoints. No significant association of the Rh matching status with any of the HSCT endpoints was seen. These results suggest that ABO minor mismatches may play a role in aGvHD pathophysiology, possibly by providing the setting for T cell activation and antibody mediated damage. To decrease the risk of aGVHD, ABO matching should be considered in HSCT.

Purpose: To determine whether corneal graft survival over a 5-year follow-up period was affected by ABO blood type compatibility in participants in the Cornea Donor Study undergoing corneal transplantation principally for Fuchs dystrophy or pseudophakic corneal edema, conditions at low-risk for graft rejection., Design: Multi-center prospective, double-masked, clinical trial., Methods: ABO blood group compatibility was determined for 1,002 donors and recipients. During a 5-year follow-up period, episodes of graft rejection were documented, and graft failures were classified as to whether or not they were attributable to immunologic rejection. Endothelial cell density was determined by a central reading center for a subset of subjects., Results: ABO donor-recipient incompatibility was not associated with graft failure attributable to any cause including graft failure because of rejection, or with the occurrence of a rejection episode. The 5-year cumulative incidence of graft failure attributable to rejection was 32 (6%) for recipients with ABO recipient-donor compatibility and 12 (4%) for those with ABO incompatibility (hazard ratio, 0.65; 95% confidence interval, 0.33 to 1.25; P = .20). The 5-year incidence for a definite rejection episode, irrespective of whether graft failure ultimately occurred, was 64 (12%) for ABO compatible compared with 25 (8%) for ABO incompatible cases (P = .09). Among clear grafts at 5 years, percent loss of endothelial cells was similar in ABO compatible and incompatible cases., Conclusions: In patients undergoing penetrating keratoplasty for Fuchs dystrophy or pseudophakic corneal edema, ABO matching is not indicated since ABO incompatibility does not increase the risk of transplant failure attributable to graft rejection.

Patients whose RBCs are D- may produce anti-D if they are exposed to D on donor RBCs. Except in emergency situations, patients whose RBCs lack D are transfused with only D- RBCs. Platelets carry no Rh antigens, but platelet units may be contaminated by RBCs that could carry D when these units are collected from D+ donors. The purpose of this study was to determine whether our policy of allowing D+ platelets to be transfused to patients whose RBCs type as D-, without the use of prophylactic Rh immunoglobulin (RhIG), results in D alloimmunization. The transfusion records of all patients who received platelet transfusions from December 2004 to March 2007 were reviewed. Transfusion recipients were evaluated with pretransfusion ABO and D typings, and an antibody screen. Recipients were reevaluated in the same manner before subsequent transfusions. Transfusion records of 114 D- patients were analyzed. Overall, 104 patients received D+ platelets; 67 had repeat antibody screening after transfusion. No patients were shown to make anti-D after platelet transfusion. There was no evidence of D alloimmunization as a result of transfusion of D+ platelets in any D- patient during this study. The data do not support the practice of restricting D- patients to receiving only D- apheresis platelets, even among patients with chronic transfusion requirements. Prophylactic use of RhIG for D+ apheresis platelet transfusions in D- patients also appears to be unnecessary.

We report a case of a 67-year-old male who underwent OLT from a deceased, sex-matched donor. Two months later he developed Evans syndrome and GVHD of the skin. Donor and recipient were matched for HLA-A and -B loci in the direction of rejection but mismatched in the direction of GVHD and fully mismatched for DRB1. These mismatches were permissible for engraftment of donor T-cells but led to GVHD. Chimerism appeared restricted to the T-cell compartment. In this case, partially matched passenger lymphocytes triggered a graft versus host reaction. In addition, alloantibodies caused cytopenias that improved after immunosuppression. HLA typing was critical in confirming this rare diagnosis and elucidating its cause. Recipients of solid organs from donors that are partially matched in the direction of rejection may need to be closely monitored for GVHD., ((c) 2008 Wiley-Liss, Inc.)

Background: It has been suggested that an immune response against the high immunogenic D antigen also augments the immune response to less immunogenic red blood cell (RBC) antigens. Based on the high antibody frequency, E and K antigens can also be regarded as strong immunogens. The question is whether the immunization against E and K antigens also enhances the formation of other antibody specificities. This question is in particular relevant for patients who are currently transfused with RH- and/or K-matched RBCs., Study Design and Methods: A retrospective multicenter study analyzed FY, JK, and MNS antibodies alone and in combination with anti-E and/or anti-K. Analysis was performed for primary and subsequent antibody responses., Results: In the cohort analyzed, 5016 patients possessed 5981 antibodies. Antibodies directed to multiple blood group systems were present in 606 of the 779 (78%) patients with multiple antibodies. In 88 of 1270 (6.9%) patients, FY, JK, and/or MNS antibodies appeared simultaneous with anti-E and/or anti-K during a primary antibody response after transfusion. Patients formed antibodies to antigens in the FY, JK, and MNS systems equally often as first antibodies followed by anti-E or anti-K than as second antibodies after anti-E or anti-K were already present. Patients with anti-E and/or anti-K or with antibodies to antigens in the FY, JK, and/or MNS systems equally often formed additional antibodies during a second antibody response., Conclusion: An immune response against allogeneic RBC antigens defines good responders who readily form antibodies against other antigens. No support was found that the response against strong RBC antigens also enhances the formation against weaker antigens.

Background: The objective of this study was to identify proteins from the amniotic fluid of the most common reproductive ailment, that is, Rh(-) incompatibility. In the present study, AF from Rh(-) pregnancy in the 29-35th week of gestation was analyzed by 2-DE, and our purpose was to identify the major spots on 2D gel., Methods: Proteins from an amniotic fluid sample was separated on 2D gel and visualized after silver staining. Most of the spots were analyzed by MALDI-TOF/MS, and were identified based on the NCBI database by a peptide mass fingerprinting method., Results: Albumin, serotransferrin, haptoglobin, alpha-fetoprotein and immunoglobulin were the major proteins detected. On comparison with 2D gel of normal AF, differences were observed at the level of albumin, and newly identified Calgranulins and Neutophil-defensins were observed. Many enzymes, growth factor binding proteins, protease inhibitors and proteins related to inflammation have also been identified., Conclusions: This study demonstrates, for the first time, the potential utility of proteomics-based approaches to identify specific biomarkers like Calgranulins and Neutophil-defensins and diagnostic profiles for Rh(-) incompatibility processes and pathophysiologic conditions of pregnancy.

Immune complexes (IC) are constantly formed at low levels in normal individuals. In humans, the red blood cell (RBC) complement receptor 1 (CR1) plays the dominant role in the IC binding and clearance. Over the last several years, we have investigated the potential utility of immunocamouflaged (methoxypoly(ethylene glycol) [mPEG] grafted) RBC to attenuate the risk of alloimmunization. Because the grafted polymer nonspecifically camouflages membrane proteins, its effects on CR1 detection and IC binding were assessed. The dose dependent (0-2.5 mM) effects of activated mPEG (CmPEG, 5 kDa; and BTCmPEG, 5 and 20 kDa) on CR1 detection and the binding of artificially generated IC [C3b coated alkaline phosphatase and antialkaline phosphatase complexes] to control and pegylated RBC was investigated by flow cytometry. In contrast to selected non-ABO blood group antigens, grafted mPEG did not effectively camouflage CR1. Surprisingly, however, even very low grafting concentrations of mPEG (>or=0.3 mM) resulted in a >or=95% loss in IC binding. Further reductions in grafting concentration (0.15 and 0.03 mM mPEG) still yielded decreased IC binding of approximately 60 and 40%, respectively. Importantly, unactivated mPEG had minimal effects on IC binding. These data demonstrate that even small amounts of grafted mPEG interfere with the multivalent CR1-IC interaction necessary for high affinity IC binding, hence large volume transfusions of mPEG-RBC may be contraindicated in patients with pre-existing IC disease. Whether this concern is of clinical significance in healthy humans is less clear due to dilutional effects and the presence of secondary clearance pathways., ((c) 2007 Wiley-Liss, Inc.)

In the present day, pretransfusion tests include ABO and RhD grouping, antibody screening, antibody identification, and cross matching. Although error rates for these tests have decreased compared to those in the past, clerical errors still occur. When exposed to RhD positive RBCs, a RhD negative person can produce anti-D that causes a severe hemolytic disease of the fetus and the newborn in addition to hemolytic transfusion reactions. Therefore, administration of RhD positive RBCs to a RhD negative person should be avoided. We experienced a RhD negative patient who had been misidentified as positive and transfused 35 units of RhD positive RBCs eight years ago, but did not have detectable anti-D in present. The red cells of the patient showed no agglutination with the anti-D reagent and a negative result in the standard weak D test. The multiplex PCR with sequence-specific priming revealed that the patient was RhD negative.

Objective: Intrauterine transfusion (IUT) is a life-saving therapy for the severely anemic fetus with hemolytic disease. However, maternal additional antibody formation is a complication of the procedure. In this study, we determined antibody formation after introduction of preventive Rh-D, -C, -c, -E, and -e and K matching of IUT donors., Study Design: This was a retrospective follow-up study., Results: During an 11-year period, 686 Rhesus- and K-matched IUTs were performed in 233 pregnancies and in 95% (652/686) posttransfusion antibody testing was performed after a median interval of 21 days. Twenty-five percent (53/212) of the women formed 64 new antibodies and, compared to our previous study, this incidence was not decreased by the use of Rhesus- and K-matched donors. After delivery, 72% (153/212) of the women had multiple RBC antibodies. Additional antibodies were in 48% (31/64) directed against Rhesus and K antigens, induced by the fetus, or as natural antibodies. In 52% (33/64) the antibodies were directed against non-Rhesus and -K antigens and in 65% (11/17) of eligible cases the IUT donor and not the fetus expressed the corresponding antigen(s)., Conclusion: Despite Rhesus- and K-matching, women treated with IUTs still show strikingly broad red cell alloimmunization. More extensive IUT donor red cell matching, including FY, JK, and S antigens, to reduce the formation of new red cell antibodies should be explored.

It is well known that certain combinations of alloantibodies are frequently found together. Patients with sickle cell disease (SCD) are mostly ofAfrican ancestry,and they may make anti-hrB. A transfusion of hrB- blood is often achieved by using e- (R2R2) RBCs; it is generally believed that hrB- patients readily make anti-E or a"broad-spectrum" anti-Rh34 (-HrB). We describe two multiply transfused D+ patients with SCD and a history of anti-hrB who subsequently produced anti- D. This raises the question whether anti-hrB together with anti-D is a more common antibody combination than anti-hrB with anti-E or anti-Rh34.

Nowadays management of severe Rh alloimmunization consists of serial determination of middle cerebral artery peak systolic velocity, amniocentesis, cordocentesis, and in many instances intrauterine transfusion. We present a case of severe Rh alloimmunization who, for the first time in Iran, was delivered at term after several intrauterine transfusions.

Background: The D antigen of the polymorphic Rh blood group system is of particular clinical importance regarding transfusion- and pregnancy-induced alloimmunization. Different RhD variants with specific clinical implications have been characterized. The least expressed D variants collectively called DEL are serologically detectable only by adsorption-elution techniques, with so far only poorly defined antigenic properties., Study Design and Methods: A comprehensive immunohematologic analysis of five of the six currently known DEL genotypes was performed. DEL phenotypes associated with the RHD(M295I), RHD(IVS3+1g>a), RHD(K409K), RHD(X418L), or RHD(IVS5-38del4) allele were characterized with extended serology and flow cytometry., Results: Epitope mapping with adsorption-elution revealed a prominent D epitope loss in the RHD(IVS3+1g>a)-associated DEL phenotype, whereas in the other four DEL types no signs of qualitative D antigen alteration were detected. The observation of alloanti-D in two RHD(IVS3+1g>a) cases confirmed the partial nature of this DEL phenotype. The RHD(M295I) phenotype exhibited the highest D antigen expression among all investigated DEL types, as determined by a semiquantitative adsorption-elution approach and flow cytometry., Conclusion: In conclusion, evidence is provided that different DEL genotypes code either for partial or complete D antigen expression and that this finding is clinically relevant.

Background: The D antigen includes category D, partial D, and weak D types, which are important because anti-D alloimmunization can occur in some but not all persons that express a variant RHD allele. At present, there is little prospective information on the prevalence of D variants among obstetric patients and potential transfusion recipients., Study Design and Methods: The RHD alleles were prospectively examined in a large patient population identified on the basis of a difference in anti-D reactivity between two reagents., Results: Fifty-five discrepancies (0.96% of D-) were noted among 33,864 ethnically diverse patients over 18 months, of which 54 represented mutated RHD alleles. Seven obstetric patients were assigned D- status based on serology; only 1 patient had a partial RHD allele. Ten of 25 (36%) obstetric patients and 4 of 6 (67%) female potential transfusion recipients of childbearing age or younger were assigned D+ status, and they expressed a D variant known to permit anti-D alloimmunization. In total 20 RHD alleles were identified including category, DVa or DVa-like alleles (n = 7), DAR (n = 8), and four novel RHD alleles including two new DAU alleles., Conclusion: Given the complexity of D antigen expression, it is concluded that some clinically important D variants identified by standard serologic analysis phenotype as D+ and are potentially at risk for the development of anti-D.

Introduction: Guidelines for distribution and use of blood products have been established for both blood transfusion institution and hospitals, in particular for the use of Rh (D)-incompatible platelet concentrates. The aim of this study was to evaluate: 1) the rate of attribution for the Rh (D)-incompatible platelets concentrates, 2) the immunisation prophylaxis practices, 3) the immunological consequences using short and medium term follow-up of transfused patients., Methods: Patients with Rh (D)-incompatible platelets concentrate administered during the year 2003 at Rouen University Hospital were retrospectively selected. Patients on transfusion were described. The relationship of various factors with the injection as well as the appearance of allo-immunization was statistically tested., Results: During a year, 280 Rh (D)-incompatible platelets concentrates were administered to 67 patients. Immunisation prophylaxis by injection of Ig anti-D was not systematically performed. Four immunizations in the Rhesus group system were identified: 2 against D antigen (Ag), 1 against E Ag and 1 against C Ag. Immunisations against D Ag occurred for two younger women considered as immunodeficient. Immunization prophylaxis was more frequent in poly-transfused patients. However no difference was observed for the other factors., Conclusion: Compatibility concerning Rhesus (D) is not always possible. The immunization against red cells persists, in particular against the antigens of the Rhesus group system and moreover for the immunodeficient patients. Recommendations for immunization prophylaxis by injection of specific anti-D immune-globulin (Ig) could be reconsidered.

This is a case of a patient, who had peripheral blood progenitor cells transplantation as treatment for aplastic anaemia and changed her blood group phenotype from Rh plus to Rh minus. When she became pregnant, she produced very early anti-Rh-D antibodies. These antibodies were directed against erythrocytes of the foetus who inherited Rh-D antigen after his mother (father was Rh minus). During this pregnancy we observed a mild example of Rh-disease, which required fetal blood testing, without the necessity of intrauterine blood transfusion. We can assume, that the immunization with Rh-D antigen had taken place before pregnancy, probably during the blood group conversion after the transplantation. We think, that blood progenitor cells transplantation, different in groups of blood may exemplify a new source of immunization and development of serological conflict in pregnancy. In cases of pregnancies, in which the child's mother or father is after grafting of blood progenitor cells, when analyzing the blood group of the foetus we should know the blood groups of parents before transplantation.