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Mature cystic teratomas (MCTs) have a rare but recognised association with IgG-mediated autoimmune haemolytic anaemia (AIHA). We present the first case of an MCT associated with IgA-mediated AIHA. We discuss the diagnostic challenges of patients presenting with a direct antiglobulin test negative AIHA picture and the importance of reviewing these cases with the local transfusion laboratory. We also review the treatment options for MCT associated AIHA. In particular, we note they are typically resistant to immunosuppression and that surgical resection should be considered at an early stage. Although MCTs are a rare cause of AIHA, they can be treated easily and should be excluded in young women presenting with AIHA, especially if resistant to corticosteroids.

A 27-year-old woman presented with a history of excessive hair loss, loss of appetite, loss of weight, amenorrhoea and loss of axillary and pubic hair for 6 months followed by fever and vomiting for 5 months and abdominal pain for 1 month. During the course of her illness, the patient developed intravascular haemolysis as evidenced by a drop in haemoglobin, indirect hyperbilirubinaemia, raised lactate dehydrogenase (LDH) and haemoglobinuria. Examination revealed severe pallor, mild icterus, elevated jugular venous pressure, generalised lymphadenopathy and hyperpigmentation. Investigations revealed severe anaemia, indirect hyperbilirubinaemia, raised LDH and negative Coombs test. Antinuclear antibody and anti-dsDNA, anti-Sm and anti-SS-A/Ro antibodies were positive and complement C3 was low. The patient was diagnosed to have systemic lupus erythematosus and immune-mediated intravascular haemolysis and was treated with prednisolone and hydroxychloroquine. Haemolysis resolved following steroid therapy, and during follow-up, there were no further episodes of haemolysis.

There are few case reports of hemolytic disease in fetuses and newborns (HDFN) caused by alloantibodies against the MNS blood group system. The reason for this dearth is that antibodies toward these antigens are usually IgM, which not only cannot cross the placental circulation but also react at temperatures below 37°C. They are, therefore, of minimal clinical importance. Nevertheless, cases have been reported in which the presence of anti-M IgG antibodies caused severe HDFN and even intrauterine death in the presence of maternal-fetal MNS incompatibility indicating that they could have a high clinical impact. The hemolytic pattern observed in these cases is similar to that caused by anti-Kell antibodies. Progressive anemia is mediated and developed through hematopoietic suppression inducing the destruction of bone marrow precursor cells with the resulting absence of reticulocytes in peripheral blood. This occurred in the case of a woman at 38.5 weeks of gestation who showed a discrepancy between direct and reverse blood type determination. A direct Coombs test was performed on the newborn’s blood, which was positive in the absence of maternal-fetal ABO incompatibility. Further tests were performed and anti-M antibodies were found in the maternal serum screening. Our final diagnosis was largely due to discrepancy issues in maternal blood. Although anti-M antibodies do not usually play a significant role in HDFN, this case stresses the importance of identifying the presence of antibodies that can be crucial in preventing HDFN and lead to new recommendations for the screening and prompt treatment of hemolysis in newborns.Hay pocos reportes de enfermedad hemolítica del feto y del recién nacido causada por aloanticuerpos contra el sistema de antígenos MNS, especialmente, porque los anticuerpos que se generan contra estos antígenos son del tipo IgM, los cuales tienen reactividad a temperaturas inferiores a los 37 °C, y, por lo tanto, no son de importancia clínica. A pesar de ello, se han reportado casos con presencia de anticuerpos anti-M de tipo IgG causantes de la enfermedad hemolítica del recién nacido e, incluso, casos de muerte intrauterina por incompatibilidad materno-fetal en el sistema MNS. El proceso hemolítico se asemeja al causado por los anticuerpos anti-Kell, con anemia progresiva por supresión hematopoyética que induce la destrucción de precursores hematopoyéticos en la médula ósea y ausencia de reticulocitos en la periferia. Se reporta el caso de una mujer con 38,5 semanas de gestación, que presentó discrepancia en la hemoclasificación directa y en la inversa. Como resultado, el recién nacido fue positivo en la prueba de Coombs directa sin que existiera incompatibilidad ABO con la madre. La correlación de estos resultados llevó a la detección de un anticuerpo anti-M en el suero materno. El diagnóstico definitivo fue posible gracias a la discrepancia en la hemoclasificación de la sangre materna. A pesar de que los anticuerpos anti-M usualmente no desempeñan un papel importante en la enfermedad hemolítica perinatal, este caso resalta la importancia de determinar la presencia de diferentes anticuerpos que pueden ser de interés a la hora de prevenir resultados graves asociados con dicha condición.

The DiegoA 39-week gestation male newborn of Han nationality was delivered by second cesarean section because of scarred uterus. The newborn's birth weight was 3700 g with an Apgar score of 9. Four hours after delivery, transcutaneous bilirubin test revealed a level of 17 mg/dl. After 23 hours, the neonate developed anemia and hyperbilirubinemia. Bacterium, virus and other pathogens, as well as tests for arcuate and glucose-6-phosphate dehydrogenase, were all negative. Direct antiglobulin test of the neonate was positive. DiegoIt is important to consider and test for the anti-Diego

Summary Objectives To evaluate the potential of the automated titre score (TS) as an alternative method to continuous flow analysis (CFA) for the prediction of the nature of anti‐D in pregnancy. Background The 2016 revised British Society for Haematology (BSH) antenatal guidelines recommended a measurement of anti‐D concentration by CFA to ensure the detection of potential immune anti‐D. Due to high referral costs and resource pressures, uptake has been challenging for hospital laboratories. Serious Hazards of transfusion (SHOT) data have previously shown that this has contributed to missed antenatal follow ups for women with immune anti‐D and neonates affected by haemolytic disease of the fetus/newborn. Methods/Materials In this multicentre comparative study, samples referred for CFA quantification were also tested by an ORTHO VISION automated anti‐D indirect antiglobulin test (IAT) serial dilution and then converted to TS. CFA results and history of anti‐D prophylaxis were used to categorise samples as passive or immune, with the aim of determining a potential TS cut‐off for CFA referral of at risk patients. Results Five UK National Health Service (NHS) trusts generated a total of 196 anti‐D TS results, of which 128 were classified as passive and 68 as immune. Diagnostic testing of CFA and TS values indicated a TS cut‐off of 35 to assist in distinguishing the nature of anti‐D. Using this cut‐off, 175 (89%) results were correctly assigned into the passive or immune range, giving a specificity of 92.19% and a negative predictive value of 91.47%. Conclusion TS in conjunction with clinical and anti‐D prophylaxis history can be used as a viable and cost‐effective alternative to CFA in a hospital laboratory setting.

BACKGROUND Plasma transfusion is a critical treatment in managing bleeding patients. In an effort to make plasma immediately available in spite of the limited amount of AB plasma, providers have begun using A plasma in life-threatening emergencies. As this practice becomes widely adopted it is important to evaluate safety. Hemolytic transfusions reactions are underreported, and hemolysis may be subclinical. STUDY DESIGN AND METHODS A retrospective study was performed at the University of Florida/Shands Hospital of B and AB patients who received 1 unit or more of A plasma. Patient charts were reviewed and data collected included age; sex; mortality; intensive care unit (ICU) length of stay; and laboratory tests used in identifying hemolysis including direct antiglobulin test, lactate dehydrogenase, haptoglobin, indirect bilirubin, aspartate aminotransferase, urinalysis, hemoglobin, and hematocrit. The primary end points of the study were immune mediated hemolysis, mortality, and length of ICU stay. RESULTS Ninety-three patients were identified as eligible for the study. One patient suffered a delayed hemolytic transfusion reaction determined to be due to an anti-Jka . No evidence of hemolysis due to ABO-incompatible plasma transfusion was identified. The volume of A plasma transfused was found to be weakly related to mortality and ICU stay. CONCLUSION No evidence of ABO immune-mediated hemolysis was observed in the patient population. The results of the study support the safety of A plasma transfusion in B and AB patients. We hypothesize the relationship observed between A plasma volume and mortality/ICU stay may be from collinearity with disease severity.

of the study is to research the epidemiological aspects of maternal alloimmunization against erythrocyte antigens of fetuses (AB0, Rhesus, Lewis, Kell, Duffy and others) and to identify the most common types of hemolytic disease of the newborn (HDN) in the West Herzegovina region.The 20-year retrospective epidemiological study includes all pregnant women who had been immunologically tested and newborn treated for HDN.The indirect antiglobulin (IAT) detected antibodies against antigens in 545 (1.8%) pregnant women of the 29 663 who were tested at the Department of Transfusion Medicine. During the 20-year-long study 310 (1.0%) newborn with HDN were treated. Our results indicate that 42% (230/545) of the pregnant women had AB0 immunization. The most common form of HDN is AB0 HDN 64% (199/310), whereas RhD HDN was treated in 19% (59/310) of the newborn infants. ETR was performed on 29 (19%) infants, 21 (72.4%) with AB0 HDN, and 7 (26%) with RhD HDN.This 20-year-long study concludes that, even though there has been significant progress in the prevention of immunization and proactive treatment of HDN, precautionary measures are still required as is the need for gynecologists and obstetricians to be active. The reasons for this are the non-existence of preventive measures for non-RhD immunization, the irregular immunological screening of RhD positive women in pregnancy in the region encompassed by the study in the past few years. The above raises new questions and recommends further research and monitoring of immunization and HDN treatment worldwide.ZIEL DIESER STUDIE: war die Untersuchung von epidemiologischen Aspekten mütterlicher Immunisierung gegen fetale Erythrozytenantigene(AB0, Rhesus, Lewis, Kell, Duffy und andere) und die Feststellung von häufigsten Ursachen der hämolytischen Erkrankung des Neugeborenen (MHN) in der Region der West-Herzegowina.20-jährige retrospektive epidemiologische Studie umfasste alle Schwangeren, die immunologische getestet sowie Neugeborene, die wegen MHN behandelt wurden.Durch den indirekten antiglobulin Test (IAT) wurden bei 545 (1,8%) Schwangeren Antikörper gegen Antigene nachgewiesen, von 29 663 Probanden, die an der Transfusionsanstalt UHC Mostar getestet wurden. Während der zwanzigjährigen Studie hatten 310 (1,0%) Neugeborene MHN. Unsere Ergebnisse zeigten, dass 42% (230/545) der Schwangeren die AB0-Immunisierung hatten. Die häufigste MHN bei dieser Forschung war die AB0 MHN 64% (199/310), während RhD MHN bei 19% (59/310) der Neugeborenen behandelt wurde.Die Blutaustauschtransfusion (ETR) erfolgte bei 29 (19%) Neugeborenen, 21 (72,4%) bei Neugeborenen mit AB0 MHNund 7 (26%) mit Rh D MHN.Obwohl es zu während dieser 20-jährigen Studie zu einem bedeutenden Fortschritt bei der Vorbeugung der Immunisierung und der proaktiven MHN-Behandlung kam, erfordert es weiterhin Vorsichtsmaßnahmen und die Aktivitäten der Gynäkologen und Neonatologen. Grund dafür ist das Nichtbestehen von Vorbeugungsmaßnahmen zur nicht-RhD Immunisierung (67% der Schwangeren hatten Nicht-RhD Antikörper), unregelmäßiges immunologisches Testen von RhD-positiven Schwangeren in den letzten Jahren im Untersuchungsgebiet. All dies eröffnet neue Fragen und Empfehlungen zur Erforschung und Beobachtung der Immunisierung sowie MHN-Behandlung auf der globalen immunologischen Szene.

A 61-year old female with dengue haemorrhagic fever developed anaemia and rising transaminase levels on the 6th day of illness. She was found to have haemolysis with a negative direct antiglobulin test (DAT), and no red cell fragmentation. She recovered with supportive care. Haemolysis with associated Haemolytic uraemic syndrome (HUS), Disseminated intravascular coagulation (DIC) and cold Auto-immune haemolytic anaemia (AIHA) is reported previously; DAT negative haemolytic anaemia associated with dengue fever has not. This case suggests a need for further studies on other immune mechanisms that can lead to haemolysis with dengue fever.

Objective: We aimed to assess the frequency distribution of of ABO haemolytic disease of newborn (ABO-HDN) and to know the predictive value of immunohaematological tests in identifying at risk neonates. Background: ABO incompatibility, although a common cause of haemolytic disease of newborn, has several unaddressed issues related to it. Material and methods: A prospective study over 20 months was carried out in a tertiary care centre in South India. Blood grouping, Direct Antiglobulin test (DAT) and elution studies were performed on neonatal samples, whereas blood grouping, antibody screening and antibody titration were performed on maternal samples. In suspected cases, ABO-HDN was diagnosed after excluding other possible causes for haemolysis. The laboratory results were correlated with the clinical details to assess the predictive value of the tests. Results: Of the total 2856 pregnancies, 34% had ABO incompatibility. On testing with columnagglutination test (CAT), the overall DAT positivity and that among ABO-incompatible cases were 3.8% and 11.2%, respectively,) whereas by conventinal tube technique (CTT) it was 0.6% and 2.4% respectively. CAT was found to have higher sensitivity, and the predictive value was higher for CTT. Maternal IgG titre showed a positive linear relationship with the DAT strength and the rise in indirect bilirubin levels. The positive predictive value of combination of tests such as DAT, elution and titation was 94.12%, which was much higher than that of the individual tests. Conclusion: DAT positivity is a predictor of early rise in serum bilirubin level, and a combination of tests has a better predictive value than individual tests towards development of clinically significant hyperbilirubinemia in ABO-HDN.

ALX148, a novel CD47 blocking agent, is in clinical development for the treatment of advanced solid tumors and lymphoma. Because CD47 is highly expressed on red blood cells (RBCs), its therapeutic blockade can potentially interfere with pretransfusion compatibility testing. This study describes the interference of ALX148 in pretransfusion compatibility testing and evaluates the methods used for mitigating such interference. Study design and methods Routine serologic tests were performed on six samples from four patients treated with ALX148. Antibody screening tests were performed on ALX148-spiked plasma, and RBC testing including antigen typing was performed on ALX148-coated RBCs. Soluble CD47 or high-affinity signal regulatory protein α (SIRPα) monomers were used to remove the false-positive reactivity of ALX148-spiked plasma with or without anti-E. Results ALX148 caused false-positive reactivity in antibody screening using indirect antiglobulin testing (IAT) and two-stage papain testing. However, false-positive reactivity was not observed at the immediate spin (IS), room temperature (RT), and 37°C phases. Direct antiglobulin testing, autologous controls, and eluates showed positive results. ALX148 did not affect blood group antigen typing performed at the IS or RT phases. The use of 50- to 100-fold molar excess of soluble CD47 or 300-fold molar excess of high-affinity SIRPα monomers removed false-positive reactivity in IAT without affecting anti-E detection. Conclusion ALX148 generates false-positive reactivity in IAT, interfering with pretransfusion compatibility testing. The use of soluble CD47 or high-affinity SIRPα monomers can resolve the interference without possibly missing clinically significant alloantibodies.

This study investigated a person with an AB0 discrepancy. Her blood group initially typed at the birth as AB Rh+ (positive); however, it was B Rh+ (positive) or Rh- (negative) when she was in her teens. At room temperature, her erythrocytes were agglutinated by anti-B, and the agglutination was significantly weaker at 37amp;ordm;C. As a result, her erythrocytes did not absorb anti-B but anti-A. Furthermore, her erythrocytes were agglutinated by anti-A at 37amp;ordm;C with signs of hemolysis in the presence of complement. The unwashed erythrocytes were also agglutinated in an antiglobulin test by polyclonal anti-A at 37amp;ordm;C and by heated polyclonal anti-A and anti-A MAB 2-8 at room temperature. Moreover, her serum agglutinated A erythrocytes at room temperature with less activity at 37amp;ordm;C; however, it agglutinated B erythrocytes at 37amp;ordm;C. The ability of the erythrocytes of this person to absorb anti-A came along with the agglutination of her erythrocytes at 37amp;ordm;C by polyclonal serum and decreased activity of the serum to agglutinate A erythrocytes at 37amp;ordm;C, compared to room temperature. The absence of anti-B absorbance by the personamp;rsquo;s erythrocytes was accompanied by the presence of anti-B in the serum, which was active at 37amp;ordm;C. The incubation of the personamp;rsquo;s serum with 0 erythrocytes induced the ability of erythrocytes to absorb anti-A and to be hemolyzed by anti-A in the presence of complement in accordance with the personamp;rsquo;s characteristics of erythrocytes. The reaction of absorption and agglutination at room temperature and 37amp;ordm;C by heated serum with the use of complement may help to reveal both weak A and B antigens and anti-A and anti-B antibodies while AB0 blood typing.

Background Neonatal jaundice is a common finding in newborns in Asia, including Indonesia. In some cases, the serum total bilirubin levels exceeds the 95th percentile for hours of life (neonatal hyperbilirubinemia). Severe neonatal hyperbilirubinemia (NH) could lead to kernicterus and neonatal death. Glucose-6-Phosphage Dehydrogenase (G6PD) genetic variations and deficiency have been reported in several studies to be associated with NH. This study aimed to analyze the G6PD genetic variations and its activity in neonates with and without hyperbilirubinemia in the Deutromalay Indonesian population. Methods Deoxyribose Nucleic Acid (DNA) was isolated from peripheral blood of 116 and 115 healthy term neonates with and without hyperbilirubinemia. All infants underwent the following laboratory examinations: routine hematologic evaluation, Coombs test, G6PD activity measurement using the Randox kit method, and serum total bilirubin level. All exons of the G6PD gene were targeted for deep sequencing using MiSeq (Illumina). An association study of G6PD polymorphisms with NH was performed using PLINK. Results The prevalence of G6PD deficiency in neonates with and without hyperbilirubinemia in Indonesian Deutromalay population were 1.72% (95% Confidence Interval (CI): 0.6–4.1%) and 1.74% (95% CI: 0.7–4.1%), respectively. The most common G6PD polymorphisms, i.e. rs1050757/c.* + 357A > G, rs2230037/c.1311C > T, and rs2071429/c.1365-13 T/IVS11, were identified. However, none of those polymorphisms and their haplotype were associated with NH (p > 0.05, Odds Ratio (OR) ~1.00). The prevalence of G6PD mutations in neonates with and without hyperbilirubinemia were 6.8% (95% CI: 2.3–11.5%) and 6.9% (95% CI: 2.3–11.6%), respectively. The most frequently identified G6PD mutation was the Viangchan variant (p.V291 M), which was followed by the Canton (p.R459L) and Vanua Lava (p.L128P) variants. Two novel mutations were identified both in case (p.V369A, p.I167F) and control (p.L474=, p.I36T) groups. Conclusion The prevalence of G6PD deficiency is low in neonates with or without hyperbilirubinemia in Deutromalay Indonesian population. The majority of G6PD mutations identified among Indonesian Deutromalay population in this study are Viangchan, Canton and Vanua Lava variants.

Background Paroxysmal nocturnal hemoglobinuria (PNH) is a rare disease presenting with variable and various clinical findings. PNH might be overlooked and diagnosis may be delayed due to low awareness about PNH. This is the first multicenter study in Turkey, investigating the efficiency of diagnostic screening of PNH by multiparameter flow cytometry (FCM) according to consensus guidelines. Methods We evaluate the efficiency of consensus clinical indications for PNH testing with FCM in 1689peripheral blood samples from 20 centers between January 2014 and December 2017. Results Overall, at the 20 centers contributing to this study, PNH clone were detected in 62/1689 samples (3.6%) by FCM test. 75.8% (n = 47) of patients with PNH clone had aplastic anemia, 3.2% (n = 2) had Coombs (-) hemolytic anemia, 6.5% (n = 4) had unexplained cytopenia, 3.2% (n = 2) had MDS with refractory anemia, 1.6% (n = 1) had hemoglobinuria and 9.7% (n = 6) had others (elevated LDH, splenomegaly, etc.). In contrast, we detect no PNH clone test in patients who were screened for unexplained thrombosis. Conclusions Our study showed that current clinical indications for PNH testing are highly efficient and diagnostic screening of suspected patients for PNH with FCM is recommended. However, advanced screening algorithms are required for patients presenting with unexplained thrombosis and normal complete blood count.

We report on a term infant with clinically significant hemolysis and hyperbilirubinemia. Testing revealed ABO incompatibility between maternal type A and infant type AB. The maternal alloantibody screen was negative. The infant’s direct antiglobulin test was positive, and anti-B IgG was eluted off the infant’s red blood cells (RBCs). Testing of the mother’s plasma revealed a high anti-B titer. The infant was successfully treated with phototherapy and intravenous immunoglobulin. The bilirubin and hematocrit stabilized, and the infant was discharged home. This case was unusual because of its severity and unusual ABO constellation. Furthermore, this report is an exemplary educational case study on how effective collaboration between the clinical team and the blood bank laboratory is critical in reaching the correct diagnosis. In summary, the differential diagnosis of more unusual and atypical ABO-incompatible constellations must be considered when an infant presents with unexplained hemolysis.

To study the serological detection characteristics and antibody specific distribution of hemolytic disease of the newborn (HDN) caused by irregular antibodies through retrospective case analysis.A total of 3 047 suspected cases of HDN were submitted by the Neonatal Department of our hospital from January 2014 to December 2019. Non ABO-HDN cases confirmed in our laboratory were taken as the research objects, while some cases of ABO-HDN were randomly selected as control. Disease-causing antibody specificity, serological detection characteristics, total bilirubin change trend and gender ratio of non ABO-HDN patients were explored.Sixty-seven cases of non ABO-HDN were confirmed from the suspected cases of HDN, Among which 45 males and 22 females were detected with the positive rate 1.48% and 0.72%, respectively. The mothers of 65 cases had two or more pregnancies. The detected irregular antibodies were mainly involved with Rh system, MNS system, Kidd system and Lewis system, among which Rh system accounted for 88.07% of the total antibody detection rate. Compared with that of ABO-HDN patients, the total bilirubin of non ABO-HDN patients developed more rapidly with a higher peak and a longer duration (P0.001). In terms of serological detection, the positive rate of non ABO-HDN direct antibody test was 97.01%, which was higher than 47.00% of ABO-HDN (P0.001), and the agglutination strength was often ≥ 2+, but there were still weak positive or negative cases of direct antibody test.Non ABO-HDN caused by irregular antibodies mostly occurs in fetuses whose mothers experience multiple pregnancies, and the number of males is more than females. The irregular antibodies detected are mainly attributed to Rh system. The peak value of bilirubin in non ABO-HDN patients is higher and lasts longer than that in ABO-HDN patients. Direct antiglobulin test may be used to roughly distinguish ABO-HDN from non ABO-HDN.非ABO新生儿溶血病的调查分析.通过回顾性病例分析，研究不规则抗体引起的新生儿溶血病（HDN）的血清学检测特点和抗体特异性分布.选取2014年1月至2019年12月本院新生儿科送检的疑似HDN病例3 047例，以本实验室检测证实为非ABO-HDN的病例为研究对象，同期检测标本中随机选取的ABO-HDN病例作为对照，分析致病抗体特异性、血清学检测特点、总胆红素变化趋势以及患儿性别构成比.送检病例中67例确诊为非ABO-HDN患儿，其中男性患儿45例，阳性率1.48%；女性患儿22例，阳性率0.72%。65例患儿母亲有≥2次的妊娠史。检出的不规则抗体主要涉及Rh系统、MNS系统、Kidd系统和Lewis系统，其中Rh系统最多，抗体检出率88.07%。与ABO-HDN患儿相比，非ABO-HDN患儿的总胆红素水平发展较迅速，峰值较高（P0.001），且持续时间较长。在血清学检测方面，非ABO-HDN直抗试验阳性率97.01%，高于ABO-HDN直抗试验阳性率47.00%（P0.001）,且凝集强度常常≥2+，但仍存在直抗弱阳性或阴性的情况.不规则抗体导致的非ABO-HDN大多发生在母亲有多次妊娠史的胎儿，且男性患儿多于女性患儿，检出的不规则抗体以Rh系统为主。与ABO-HDN患儿相比，非ABO-HDN患儿的胆红素峰值较高且持续时间长。直抗试验结果或能用于粗略区分ABO-HDN和非ABO-HDN.

Purpose To evaluate the clinical and genetic characteristics of 3 children with Haploinsufficiency of A20 (HA20). Methods:The clinical and genetic testing data of 3 children with HA20 treated at Capital Institute of Pediatrics (CIP) between August 2016 and October 2019 were retrospectively analysed. Result Patient 1 presented with arthritis and inflammatory bowel disease, patient 2 presented with axial spinal arthritis and lupus-like syndrome, and patient 3 presented with recurrent oral ulcers, gastrointestinal ulcers, and perianal abscesses. Regarding laboratory tests, patients were found to have elevated white blood cell (WBC) count, C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR). The CRP and ESR was reported to be high in all the patients. The WBC was reported to be high in patient 1 and 3. Patient 2 was positive for antinuclear antibodies, anti-Sjögren’s syndrome antigen A, dsDNA, rheumatoid factor and Coombs test. Genetic testing showed that all three patients had heterozygous mutation in TNFAIP3 gene. As for the treatment, patient 1 was treated with TNFα antagonist, patient 2 was treated with TNF α antagonist and sulfasalazine, and patient 3 was treated with corticosteroids and thalidomide. Patients 1 and 2 were followed for four and 3 months, respectively. There was an improvement in joint and gastrointestinal symptoms; inflammatory indices and rheumatoid factor (RF) were normal, and dsDNA and Coombs test became negative. Patient 3 was treated at another hospital and showed gradual improvement in oral ulcers and perianal abscesses. Conclusion HA20 is a single-gene auto-inflammatory disease caused by mutation in tumour necrosis factor (TNF)-α-induced protein 3 (TNFAIP3) gene. It may present as Behçet-like syndrome and resemble various other autoimmune diseases as well. Corticosteroids and immunosuppressive agents are effective treatments, and cytokine antagonists can be used in refractory cases. Whole-exome genetic testing should be proactively performed for children with early-age onset or Behçet-like syndrome to achieve early diagnosis and accurate treatment.

Berzuini et al report the observation that nearly half of patients with COVID-19 tested at their blood center had a positive direct antiglobulin test (DAT). However, eluates did not react with any test cells but did react with red cells from other patients with COVID-19 that were DAT negative. This suggests that COVID-19 may modulate the red cell membrane and present novel antigenic epitopes.

Background The diagnosis of autoimmune hemolytic anemia (AIHA) can be challenging since the direct antiglobulin test (DAT) has been reported to be falsely negative in 3%-11% of cases. In children with anemia, laboratory and/or clinical evidence of hemolysis and a negative DAT, clinicians should consider further specialized testing to confirm AIHA to accurately diagnose and treat this uncommon pediatric entity. Study design and methods A retrospective chart review was undertaken at a large tertiary care academic pediatric hematology practice to describe our experience with DAT-negative AIHA. Results From January 1, 2010 through August 1, 2016, 10 children were described who had clinical and laboratory evidence of AIHA, a negative DAT, and further specialized serologic testing confirming this diagnosis. Conclusion This case series highlights the need for further serologic workup when a child's clinical presentation is highly consistent with AIHA despite a negative DAT.

Hemolytic disease of the fetus and newborn (HDFN) is a clinically significant problem that may potentially affect any pregnancy. Direct antiglobulin test (DAT) is considered to be an important test in identifying newborns who are suspected to have HDN. This study aims in reviewing data regarding a positive DAT result concerning etiology and the development of HDN over a period of 10 years.A retrospective study of all neonates with a positive DAT result between January 2011 and December 2020 was performed. Data were obtained from patients' electronic hospital files, transfusion medicine databases, and medical birth records. Laboratory parameters along with clinical interventions in neonates with a DAT-positive result and a comparison group of DAT-negative neonates were performed.36,000 deliveries were registered in this period. 176 (2.65 %) neonates had a positive DAT result. ABO-incompatibility was the most common cause with 59.1 %; Rh incompatibility 13.8 %, minor blood group incompatibility, and other RBC-related antibodies 10.1 %, and unspecified etiology in 17 % of cases. Among DAT-positive cases, 32.7 % of neonates were diagnosed with HDN. ABO-incompatibility was the major reason as well. Initial mean total bilirubin levels were higher in the DAT-positive group than the control group (p0.001), and these neonates also had a lower initial hemoglobin level (p0.001). The need for therapeutic interventions was significantly higher in DAT-positive neonates (p0.001) as 86.8 % underwent phototherapy, with 32.7 %, and 17.6 % receiving exchange transfusion (ET) and intravenous immunoglobulin (IVIG), respectively.In conclusion, ABO incompatibility was the most common cause for neonatal DAT positivity. Besides the common causes of DAT positivity, there would be rare but important conditions that may lead to a positive result, such as antibodies passively acquired from mothers in the context of alloimmunizations or using drugs. In addition, as a high rate of therapeutic intervention was identified among neonates with a DAT-positive result, there is a crucial need for increasing awareness regarding early diagnosis of the condition, careful monitoring, and the employment of prenatal alloimmunization screening tests.

OBJECTIVE Systemic lupus erythematosus (SLE) presents with nonspecific signs and symptoms that are also found in other conditions. This study aimed to evaluate manifestations at disease onset and to compare early SLE manifestations to those of diseases mimicking SLE. METHODS Academic lupus centers in Asia, Europe, North America, and South America collected baseline data on patients who were referred to them during the previous 3 years for possible SLE and who had a symptom duration of

Detection of blood group antibodies is a crucial step for blood transfusion recipients and pregnant women to prevent potentially fatal haemolytic reactions. Due to the short, non-bridging structure of such antibodies (IgG), the indirect antiglobulin test (IAT) is required, complete with a thermal incubation phase. This incubation step, where the sample must be heated to 37 °C for several minutes, has hitherto prevented chip- and paper-diagnostics from performing a complete IAT and instead required the IAT to be performed away from the patient beside in a laboratory setting with specialist equipment – significantly delaying blood transfusions. With recent laser technology for immunohaematology, a single blood droplet can be heated. This study presents a simple diagnostic where a single 15 μL droplet sits on hydrophobic PTFE film and is heated by laser. The result of the test is then determined via placement of a paper strip where passive wicking and filtration of the sample separates positive from negative results. We demonstrate that this diagnostic can accurately and sensitively detect blood group antibodies, with results quickly read by eye without further specialist equipment or training, with potential to lead to a point-of-care antibody screen.

Background Primary cold agglutinin disease (CAD) is a monoclonal antibody (M-protein) and complement-mediated chronic hemolytic disease process. Antibody glycosylation can play a role in both antibody half-life and complement fixation. Recently, M-protein light chain (LC) glycosylation has been shown to be associated with AL amyloidosis. We hypothesized that M-protein LC glycosylation is also associated with cold agglutinin (CA) titers and CA-mediated hemolysis. Study design and methods A cross-sectional study of patients undergoing CA titer evaluation underwent mass spectrometric analysis for M-proteins and M-protein LC glycosylation. A subset of serum samples also underwent evaluation for the ability to trigger cold hemolysis in vitro. M-protein and M-protein LC glycosylation rates were compared across CA titer groups, clinical diagnosis, direct antiglobulin testing (DAT) results, and cold in vitro hemolysis rates. Results Both M-protein and M-protein LC glycosylation rates significantly differed across CA titer groups with the highest rates in those with elevated CA titers. M-protein LC glycosylation occurred almost exclusively on IgM kappa M-proteins and was significantly associated with positive DAT results and a clinical diagnosis of CAD. Cold in vitro hemolysis was demonstrated in two patients who both had a CA titer of more than 512 but there was no significant association with CA titer group or M-protein LC glycosylation status. Conclusion M-protein LC glycosylation is significantly associated with higher CA titer levels. Given the role that antibody glycosylation can play in antibody half-life and complement fixation, further studies are needed to clarify the effects of LC glycosylation within the context of CAD.

Multiple factors contribute to anemia in patients with Hepatitis B virus (HBV)related acute-on-chronic liver failure (ACLF); however, the mechanism is unclear. The purpose of this study was to evaluate the clinical significance of the direct antiglobulin test (DAT) in patients with HBV related ACLF.DAT was used to detect immunoglobulins and/or complement proteins on the surface of erythrocytes.We recruited 78 HBV-associated ACLF patients, 30 chronic hepatitis B(CHB)patients and 40 healthy people between October 2015 and May 2016. In HBV related ACLF patients, the hemoglobin concentration, number of erythrocytes, and hematocrit value were significantly lower, while the erythrocyte distribution width was significantly higher, compared to patients with CHB and healthy controls (HCs) (P0.001). The rates of DAT positivity in HBV related ACLF patients, CHB patients, and HCs were 62.8 %, 13.3 %, and 0%, respectively. DAT-positive ACLF patients exhibited lower Hb levels, older average age, as well as higher total bilirubin, alanine aminotransferase, and complement component 3 levels compared to DAT-negative patients.HBV related ACLF patients showed significant alterations in erythrocyte parameters, possibly reflecting disease development and severity. The high presence of erythrocyte autoantibodies suggested that immunologic clearance of erythrocytes contributed to multifactorial anemia in HBV related ACLF patients.

Background In December 2019, a new coronavirus (named severe acute respiratory syndrome coronavirus 2, SARS-CoV-2) spread from China, causing a pandemic in a very short time. The main clinical presentation of SARS-CoV-2 infection (COVID-19, coronavirus disease-2019) is pneumonia, but several cardiovascular complications may also occur (e.g., acute coronary syndromes, pulmonary embolism, stroke, arrhythmias, heart failure and cardiogenic shock). Direct or indirect mechanisms induced by SARS-CoV-2 could be implicated in the pathogenesis of these events. Case presentation We report herein the third case of COVID-19 autoimmune haemolytic anaemia (AIHA) reported so far, which occurredwithout any other possible explanations in a Caucasian patient. The patient also suffered from ST-elevation myocardial injury. Conclusions Both complications occurred quite late after COVID-19 diagnosis and were probably precipitated by systemic inflammation, as indicated by a significant delayed increase in inflammatory markers, including interleukin-6 (IL-6).

Introduction Abnormally activated complement system induces erythrolysis in a part of autoimmune hemolytic anemia (AIHA) patients. However, the alterations in serum complement levels in these patients are seldom reported. In this study, we aimed to evaluate the serum complement features of AIHA patients according to different clinical and laboratory characteristics and to find relationships between complement levels and hemolysis-associated laboratory indexes. Methods A retrospective analysis of 146 AIHA patients was performed, and serum complement C3 and C4 levels were compared between control subjects and AIHA patients with different subtypes. Correlations of serum C3/C4 levels with titers of cold agglutinin test (CAT), direct antiglobulin test (DAT), and serological indexes were assessed. Spearman correlation analysis was performed to analyze the relationship between serum complement levels and other laboratory indexes. Results Autoimmune hemolytic anemia patients showed reduced serum C3 levels, while serum C4 levels tended to be lower in DAT-positive AIHA patients but not in DAT-negative AIHA patients. Patients with warm AIHA secondary to connective tissue diseases and cold agglutinin disease/cold agglutinin syndrome had the lowest serum C3/C4 levels. Serum C4 levels were negatively correlated with CAT (P = .004) and DAT (anti-C3d) (P = .007) titers. In patients with positive CAT and/or DAT (anti-C3d) but negative DAT (anti-IgG), serum C3/C4 levels were negatively correlated with indirect bilirubin (P = .017 and =.026, respectively). Conclusion The study findings may be helpful in not only unraveling the mechanism underlying hemolysis in AIHA but also diagnosing AIHA and selecting targeted treatment strategies.

Background: This research evaluated the association between the mother-infant blood type or rhesus (ABO or Rh) incompatibility, the pattern of neonatal jaundice, and serum bilirubin (TSB) values obtained prior to discharge from hospital of healthy born neonates with gestational age >34 weeks and birth weight >2000 g. Methods: We utilized a laboratory and neonatal database to identify the cord blood ABO/Rh and direct antiglobulin test (DAT) and TSB measured during hospitalization and re-admission with hyperbilirubinemia for phototherapy treatment. We used hour-specific TSB to analyze the TSB levels for ABO/Rh compatibility and isoimmunization using chi-square, analysis of variance, and regression models. Results: Of the 901 infants studied, 158 (17.5%) had ABO/Rh incompatibility, including 27 with positive DAT. Hyperbilirubinemia was diagnosed in 33.3% DAT positive, 6.9% DAT negative, and 4.6% of infants with compatible blood types. Increased predischarge TSB was observed in DAT positive infants at 48–72 h of postnatal age (P < 0.001). After controlling for age at TSB testing and weight loss percentage, multiple regression analysis did not show any impact of ABO/Rh incompatibility and DAT results on the predischarge TSB levels. Conclusion: Blood type incompatibility increases the frequency of hyperbilirubinemia only in the DAT-positive infants. Irrespective of the isoimmunization status, it does not significantly affect the level of predischarge TSB.

Acquired thrombotic thrombocytopenic purpura (aTTP) is a rare autoimmune disorder characterized by auto-antibodies to Willebrand factor (vWF) cleaving enzyme (ADAMTS13), resulting in unusually large vWF multimers that lead to platelet aggregation, microthrombi formation and microangiopathic hemolytic anemia. Hemolysis in aTTP is mechanical; thus, direct antiglobulin test (Coombs test) is usually negative. Multiple autoimmune conditions and various auto-antibodies have been described in the context of chronic myelomonocytic leukemia (CMML). In this paper, we describe the first case of CMML with auto-antibodies to ADAMTS13, presenting initially as plasmapheresis-refractory Coombs-positive aTTP. ----- Results: Although our patient was not treated for CMML, a complete remission of aTTP was eventually achieved with rituximab. Conclusion; We propose that aTTP should be in the differential diagnosis of CMML patients with thrombocytopenia and anemia (Coombs positive or not) who develop signs of thrombotic microangiopathy. Further studies are much needed to decipher the immune-mediated processes in CMML.

Asymptomatic VL is a concern, considering the risk of transmission in highly endemic areas due to human-to-human transmission. The aim of this study was to report the sero-epidemiological prevalence in Bihar, India, a highly endemic area of VL, using the leishmanin skin test (LST) and the direct agglutination test (DAT). This was a cross-sectional study performed in Muzaffarpur, Bihar, India. Relatives of patients with VL were tested by LST and DAT. Other epidemiological data were evaluated and correlated with tests results. Forty individuals (either previous or current patients), and 109 household contacts were studied. There were 36% of male visceral leishmaniasis family members versus 17.57% of females visceral leishmaniasis family members, thus showing more males with symptomatic disease than females (p< 0.01). All visceral leishmaniasis cases had positive DAT tests, but only 37% of past cases were positive on the skin testing. Amongst healthy household contacts, 34% were DAT-positive, whilst 21% were LST-positive. The overall positivity for both assays combined was 44.8% and 23.8% were DAT-positive alone. The finding of high infection prevalence amongst asymptomatic individuals, and the estimation of those at greater risk for overt disease (DAT-positive alone) are important in the development of future disease control policies.

AimsPositive direct antiglobulin tests (DATs) are valuable in identifying the aetiology of autoimmune haemolysis and in guiding therapeutic intervention. However, in HIV-positive individuals with background polyclonal gammopathy, a positive DAT in the absence of haemolysis is common. In this setting, IgG quantification and subtyping may be of value, as this is possible with the recently introduced gel cards. There is paucity of literature evaluating the diagnostic usefulness of IgG subtyping and quantification in HIV-positive individuals who are investigated for autoimmune haemolytic anaemia (AIHA). This study evaluated the usefulness of IgG quantification and subtyping in the diagnostic work-up of AIHA in patients with a positive DAT, with and without HIV infection.MethodsThis retrospective, cross-sectional study included patients investigated for AIHA in a quaternary care hospital. Those with a positive DAT had their IgG subtyped and quantified using the ID-Card DAT IgG1/IgG3 and IgG-dilution cards (Bio-Rad, Cressier, Switzerland).ResultsNinety patients admitted from December 2019 to March 2020 were investigated for AIHA. Forty-four (49%) patients had a positive DAT of whom 26 (59%) had evidence of haemolysis, and 16 (36%) were HIV positive. Concurrent HIV and haemolysis were present in eight patients, two of whom had IgG1 although none had an IgG antibody titre >1:30. None of the HIV-positive patients without features of haemolysis had IgG1/IgG3 or IgG antibody titres >1:30.ConclusionIn our clinical setting, IgG quantification and subtyping were found to be of limited value in the diagnostic characterisation of AIHA in HIV-positive patients with false-positive DAT.

Background and Objectives: Drug-induced immune hemolytic anemia is a rare condition that occurs primarily because of drug-induced antibodies, either dependent or independent and positive direct antiglobulin test. Our aim was to evaluate the association of positive DAT with nonreactive eluate and DIHA. Materials and Methods: From 2014–2018, we evaluated 159 patients who presented positive DAT with a nonreactive eluate. Laboratory and clinical analyses were performed including HIV, HBV and HCV testing. All patients were exposed to the following drugs: Dipyrone in 63.5 %, Furosemide in 28.9 %, Metoclopramide in 34.6 % and Ondansetron in 41.5 %. Results: Results of DAT showed IgG in 125 (78.4 %) patients and C3d in 24 (15.1 %) with reactions varying from 1+ to 4+. HIV test was positive in 10 (16.1 %) patients, HBV was positive in 3 (4.7 %) and HCV was positive in, 1 (1.5 %). There was no clinical significance when the parameters of hemoglobin, hematocrit, reticulocytes and LDH were evaluated, only a slight increase in bilirubin, especially, in patients with positive DAT reacting 3+/4+ due to IgG and C3d sensitization. Clinical evaluations showed that all patients were asymptomatic. Conclusions: The association of drugs with positive DAT can be a challenge to transfusion services and immunohematology reference laboratories. There was no evidence of any case of severe hemolysis with clinical repercussion through the clinical and laboratory findings analyzed with the drugs associated with positive DAT. Dipyrone and Furosemide have already been associated with DIHA but there are no studies reporting the association of Metoclopramide and Ondansetron with DIHA.

BACKGROUND: Plasma of patients taking anti-CD38 monoclonal antibodies (MoAbs) leads to panagglutination in the indirect antiglobulin test (IAT), that can mask clinically significant alloantibodies. Dithiothreitol (DTT) treatment of test RBCs is the more widespread method for avoiding this interference. Current DTT 0.2 mol/L method is time consuming and damages several red blood groups antigens. This study aims to evaluate low concentration DTT treatment of RBCs adapted for gel testing. MATERIALS AND METHODS: Four DTT concentrations (0.01, 0.02, 0.03, and 0.04 mol/L), and three gel test brands were evaluated on six DARA patient’s samples. Briefly, the method consists of pipetting 50 μL of 0.8% RBCs on AHG micro columns, followed by 25 μL of DTT, thoroughly mixing and 15 min incubation at 37 °C. Then, 25 μL of serum/plasma is added to proceed to IAT. In order to asses the effect of DTT 0.04 mol/L on different blood group antigens, serial dilutions of sera containing anti-K, -k, -Kp(b), -Lu(b), -Yt(a) and anti-JMH antibodies were tested against DTT-RBCs. One sample of a DARA patient with known alloantibodies as well as samples of two patients inoculated with anti-K and anti-Fy(a) were evaluated. RESULTS: RBCs treatment with DTT 0.04 mol/L for 15 min completely eliminated anti CD38 panagglutination in all samples studied and worked with different reactivity intensities in IAT and gel brands. The new method allowed the detection of underlying anti-D, anti-E, anti-K and anti-Fy(a) alloantibodies. Titration assays demonstrated no denaturation of Kell, Lutheran, Cartwright and JMH antigens. DISCUSSION: The new DTT method adapted for gel testing is efficacious, simple and only adds 15 min over regular IAT. Pheno/genotyping before DARA treatment or transfusion of K negative RBCs may be unnecessary.

Background: Paroxysmal nocturnal hemoglobinuria (PNH) is a rare acquired hematopoietic stem cell disease that may lead to weakness and death of patients, if unrecognized and untreated. Although consensus guidelines were reviewed recently for the diagnostic screening of PNH with multi-parameter flow cytometry (FCM), until now, no study has investigated the efficiency of such clinical indications in older patients. Methods: Overall, 20 centers participated in the study and a total of 1,689 patients were included, 313 of whom were at geriatric age and 1,376 were aged 18 - 64 years. We evaluated the efficiency of consensus clinical indica-tions for PNH testing using FCM in peripheral blood samples and compared the results of older patients and pa-tients aged 18 - 64 years. Results: PNH clones were detected positive in 7/313 (2.2%) of the older patients. Five (74.4%) of the patients with PNH clones had aplastic anemia, 1 had unexplained cytopenia, and 1 patient had myelodysplastic syndrome (MDS) with refractory anemia. PNH clones were not detected in any older patients who were screened for unex-plained thrombosis, Coombs (-) hemolytic anemia, hemoglobinuria, and others (e.g., elevated lactate dehydroge-nase (LDH), splenomegaly). We detected PNH clones in 55/1376 (4%) samples of the patients aged under 65 years. Forty-two (76.4%) patients with PNH clones had aplastic anemia, 2 patients had Coombs (-) hemolytic anemia, 3 patients had unexplained cytopenia, 1 patient had MDS with refractory anemia, 1 patient had hemoglobinuria, and 6 (10.9%) had others (e.g., elevated LDH, splenomegaly). PNH clones were not detected in any patients who were screened for unexplained thrombosis. There was no statistical difference between the geriatric population and patients aged 18 - 64 years in terms of clinical indications for PNH screening with FCM (p = 0.49). Conclusions: Our results showed that the current clinical indications for PNH screening with FCM were also effi-cient in older patients. We suggest that older patients with unexplained anemia, myelodysplastic syndrome with refractory anemia, and unexplained cytopenia should be screened for PNH with FCM to identify patients who would benefit from treatment. © 2020 Verlag Klinisches Labor GmbH. All rights reserved.

Autoimmune haemolytic anaemia (AIHA) is a kind of autoimmune diseases characterized by autoantibodies which produced and secreted by abnormal activated B lymphocytes directed against red blood cells (RBC). Study reveals that about 50% AIHA mainly occurs secondary to lymphoproliferative disorders (LPD) and autoimmune diseases. In this study, we aim to explore the characteristics of patients with AIHA secondary to LPD. Fifteen patients with AIHA secondary to LPD (secondary group) and 60 with primary AIHA (primary group) were retrospectively included. Patients in the secondary group [(59.40 ± 4.74) y] were older than those in the primary group [(47.53 ± 2.30) y] (p = 0.024). Reticulocyte counts were lower for the secondary group [(134.55 ± 20.67) × 109/L] than for the primary group [(193.88 ± 27.32) × 109/L] (p = 0.09). Haptoglobin was higher in the secondary (0.75 ± 0.19) g/L than in the primary group (0.34 ± 0.05) g/L (p = 0.004). The ratio of CD3+CD4+/CD3+CD8+ was higher in the secondary (1.81 ± 0.41) than in the primary (1.05 ± 0.12) group (p = 0.025). Duration of remission was shorter in the secondary [(23.52 ± 5.20) months] than in the primary [(40.87 ± 3.92) months] group (p = 0.013). Relapse rate was higher for the secondary (33.3%) than for the primary (8.3%) group (p = 0.003). Mortality rate was higher in the secondary (33.3%) than in the primary (8.3%) group (p = 0.003). Progression-free survival was shorter in the secondary than in the primary group (p = 0.021). In conclusion, patients with AIHA secondary to LPD showed higher age at diagnosis, shorter remission time, and higher recurrence and mortality rates than did those with primary AIHA.

BACKGROUND Yta is a high frequency red blood cell (RBC) antigen, present in 99.7% of studied populations. It is extremely immunogenic, and when anti-Yta is present, provision of Yt(a-) blood is often challenging. The objectives of our study were to assess the incidence and severity of acute hemolytic transfusion reactions to Yt(a+) donor RBCs in recipients with preformed anti-Yta and to identify any patient factors associated with severe hemolytic reactions. STUDY DESIGN AND METHODS Patients with anti-Yta identified by the Red Cell Reference Laboratories of the Australian Red Cross Lifeblood over the past 20 years were included. Their transfusion records were collected via the referring laboratory to ascertain if any patients received RBC transfusion and if there was any evidence of transfusion reactions. RESULTS Fifty-two patients with anti-Yta were identified; only 12 were confirmed to have received a RBC transfusion. Nine received Yt(a+) or untyped allogeneic RBCs, including four patients who received a total of 16 indirect antiglobulin test (IAT) crossmatch incompatible, likely Yt(a+) RBCs. None of the nine patients had documented acute hemolytic reactions. CONCLUSION There are limited published data describing the clinical significance of anti-Yta . Based on our data, it appears that transfusing patients with anti-Yta using incompatible crossmatched RBCs does not pose a significant risk of an acute hemolytic transfusion reaction when the antibody reaction strength is weak ≤2+ (0-4) by IAT crossmatch. For strong examples of the antibody, in the absence of other assay data, such as the monocyte monolayer assay, Yt(a-) blood should continue to be sourced where possible.

Objectives Recently, daratumumab has been included in the therapeutic strategies for myeloma patients. This molecule is an antibody directed against CD38, strongly expressed on plasma cells. Nevertheless, as CD38 is also present on erythrocyte membrane, daratumumab interferes with immunohaematological tests, complicating the selection of compatible blood. Methods A total of 14 patients treated by daratumumab have been followed in our transfusion laboratory. Among them, 11 have been transfused. Dithiotreitol (DTT) has been used to inhibit the daratumumab's interference, in the pre-transfusion tests (irregular antibody screening and cross-match). Results The red blood cell treatment with DTT has been very efficacious to inhibit the daratumumab's interference in 13 patients out of 14. Some precautionary measures had to be taken into account, especially the pH and the storage conditions. An extended pheno/genotype was an additional security element in the selection of compatible blood. To simplify and to optimize the laboratory practices, a decisional flow chart has been written. Conclusion DTT red blood cell treatment is very useful and efficacious in the pre-transfusion tests of patients treated with daratumumab. It allows to avoid the selection of blood bags only on the basis of an extended pheno/genotype, what is more secure and more ethical with respect to other at higher risk patients. A clear decisional flow chart allows a quality assurance gait. Collaboration with physicians is essential.

Interleukin (IL)-35 is the latest member of IL-12 family, which plays an important role in other autoimmune diseases. Bone marrow mononuclear cells Coombs test‑positive hemocytopenia, also termed immunorelated hemocytopenia (IRH) is a type of autoimmune-associated diseases. The present study investigated the relationship of IL‑35 in patients with IRH. A total of 43 patients with IRH and 19 normal controls were enrolled in the current study. Serum levels of IL‑35 and IL‑17 in peripheral blood were evaluated by ELISA. Regulatory T cells (Tregs) level was detected by flow cytometry and IL‑35 subunits mRNA in Treg was determined using reverse transcription‑quantitative polymerase chain reaction: Epstein‑Barr virus induced 3 (EBI3) and IL‑12α chain p35. Effect of IL‑35 on T helper 17 cells (Th17) cells was determined by mix‑culture of IL‑35 with CD4+ T lymphocytes. Serum level of IL‑35 was decreased in untreated patients with IRH compared with remission patients (P

The objective of the study is to study the positivity of Coombs test or direct antiglobulin test (DAT) in systemic lupus erythematosus (SLE) patients and its relationship with disease's clinical and serological profile. Retrospective study of 373 SLE patients seen at single Rheumatology Unit. Epidemiological data (age, gender, age at disease onset, auto declared ethnic background and tobacco use), clinical (malar rash, photosensitivity, oral ulcers, discoid lesions, serositis, glomerulonephritis, convulsions, psychosis, hemolytic anemia, leukopenia, lymphocytopenia and arthritis), and serological profile (anti ds DNA, anti Ro/SS-A; anti La/SS-B, Anti RNP, Anti Sm, aCl (anticardiolipin) IgG, aCl Ig M, LA or lupus anticoagulant, rheumatoid factor and direct Coombs) were collected. Patients with a positive DAT were compared with the negatives. DAT was positive in 12.8% of patients and 54.3% of them had hemolytic anemia. In univariate analysis, a positive DAT was associated with hemolytic anemia (p

BACKGROUND In low-resource countries, screening for D antibodies to detect pregnancies at risk for hemolytic disease of the newborn is not routine practice. Retrospective data showed that 5.5% of Surinamese newborns of D-negative women had a positive direct antiglobulin test (DAT), indicating the presence of maternal antibodies against fetal antigens. Here, the frequency and clinical relevance of DAT positivity is evaluated. STUDY DESIGN AND METHODS Between April 2015 and June 2016, an observational, multicenter cohort study was undertaken among Surinamese newborns born to D-negative women. In newborns, the DAT was performed, and clinical outcomes between DAT-negative and DAT-positive newborns were compared. RESULTS Of the 232 evaluable newborns, 19 (8.2%) had a positive DAT, of which 11 of 15 antibody-tested newborns had D antibodies. DAT-positive newborns had lower hemoglobin levels (p = 0.02) and a trend toward higher bilirubin concentrations (p = 0.09) in the first days of life compared with DAT-negative newborns. DAT-positive newborns were admitted more frequently (p = 0.02), needed phototherapy treatment almost four times as often as DAT-negative newborns (26% vs. 7%; p = 0.008), and therapy took 2 days longer (p = 0.01). Exchange transfusions were performed in two newborns with D antibodies, both complicated with sepsis. The hospital stay was 2.5 days longer for DAT-positive newborns (p = 0.007). Overall, the prevalence of hemolytic disease of the newborn requiring treatment was 2.2% among the whole cohort of newborns. CONCLUSION We found a high prevalence of DAT positivity with substantial need for hyperbilirubinemia treatment in newborns in Suriname. These results stress the necessity for better management procedures in D-negative women.

Autoimmune hemolytic anemia (AIHA) is an uncommon entity that presents diagnostic, prognostic, and therapeutic dilemmas despite being a well-recognized entity for over 150 years. This is because of significant differences in the rates of hemolysis and associated diseases and because there is considerable clinical heterogeneity. In addition, there is a lack of clinical trials required to refine and update standardized and evidence-based therapeutic approaches. To aid the clinician in AIHA management, we present four vignettes that represent and highlight distinct clinical presentations with separate diagnostic and therapeutic pathways that we use in our clinical practice setting. We also review the parameters present in diagnostic testing that allow for prognostic insight and present algorithms for both diagnosis and treatment of the AIHA patient in diverse situations. This is done in the hope that this review may offer guidance in regard to personalized therapy recommendations. A section is included for the diagnosis of suspected AIHA with negative test results, a relatively infrequent but challenging situation, in order to assist in the overall evaluation spectrum for these patients.

Thrombotic thrombocytopenic purpura (TTP) has a devastating prognosis without adapted management. Sources of misdiagnosis need to be identified to avoid delayed treatment. We studied 84 patients with a final diagnosis of severe (

We reported a case of a woman with no past medical illness who presented with a few days’ history of fever, myalgia, arthralgia, hypochromic microcytic anaemia and thrombocytopaenia and who was nonstructural protein 1 antigen (NS1Ag)-positive. Haemolytic anaemia including full blood picture work-up revealed high reticulocyte count and haemolysis with positive direct Coombs test. She was started on prednisolone and was discharged well.

ObjectiveTo evaluate how clinical practice was affected by the change in direct antiglobulin testing (DAT) methodologies and subsequent stronger reported DAT results at our large academic medical center.MethodWe retrospectively reviewed DAT results of umbilical cord blood from infants with blood type A or B born to mothers with antibody-negative type O blood, based on records kept at the University of Alabama at Birmingham (UAB) Hospital, a 1400-bed academic medical center.ResultsWe randomly chose 50 neonates with positive DAT results who had been tested using the tube method and 50 whose testing had used the gel method. Although 86% of results with the tube method were positive microscopically, 52% and 40% of the DAT results with the gel method were 1+ and 2+ positive, respectively. Further, we observed an increase in the number of neonates treated with phototherapy who had been tested using the gel method.ConclusionWe report that DATs performed using the gel method had increased DAT strength compared with tube testing, which led to increased use of phototherapy by our clinical colleagues.

Background Rechallenge with oxaliplatin is common in the treatment of colorectal cancer and increases the risk of a detrimental oxaliplatin-induced immune reaction. Allergic reactions to oxaliplatin may be partially avoided by desensitization protocols involving immune suppressive drugs, slow administration and gradually increasing chemotherapeutic doses. However, non-IgE-mediated immunopathologic reactions to oxaliplatin remain challenging and may be potentially life-threatening. Case presentation Here we report two potentially fatal cases of type II hypersensitivity to oxaliplatin in metastatic colorectal cancer patients. Both patients manifested with severe thrombocytopenia, intravascular haemolysis, and acute kidney injury 4–6 h after oxaliplatin administration in a rechallenge setting. Serology revealed that the reactive entity for immune haemolysis was an IgG oxaliplatin-induced antibody. The course of anti-cancer treatment and severe adverse event after oxaliplatin rechallenge including diagnostic dilemma and the results of detailed routine clinical chemistry and hematology testing are described. Extended immunohaematology/serology testing revealed that the oxaliplatin-induced IgG antibody was present in the circulation prior to the onset of hypersensitivity, persisted for months and elicited cross-reactivity with other platinum agents. Conclusion Development of type II hypersensitivity reaction manifesting as a sudden onset of severe thrombocytopenia and immune haemolysis must be considered in patients treated with oxaliplatin, especially those on long-term therapy or when rechallenged. Step-wise diagnosis involves clinical presentation, detection of haemolysis in patient's blood and/or urine, evaluation of platelet count, and direct anti-globulin Coombs test.

Babies with ABO incompatibility are often advised frequent biochemical screening and prolonged hospital stay. Our primary objective of the study was to compare serum bilirubin levels at 48 h and 96 h of age in neonates with and without ABO incompatibility. Our prospective study included neonates with gestation ≥ 34 weeks, with or without ABO incompatibility (92 in each group). A direct Coombs test was performed on cord blood. The mean serum bilirubin and haematocrit levels in both groups at 48 h and 96 h were comparable. The mean reticulocyte count of babies with ABO incompatibility was, however, significantly higher. Late preterm and term neonates with and without ABO incompatibility have similar bilirubin levels and no increased risk of significant hyperbilirubinemia. Prolonged hospitalisation of these neonates appears to be unnecessary.