Frequency of the DI*A, DI*B and Band 3 Memphis polymorphism among distinct groups in Brazil

ABSTRACT Introduction: The Band 3 is a red blood cell protein that carries the Dia and Dib antigens from the Diego blood system. The SLC4A1 gene encodes Band 3; Band 3 Memphis is a polymorphism of normal Band 3 and has two variants, but only the variant II carries the Dia antigen. Objectives: Describe the frequencies of the DI*A and DI*B alleles and the Band 3 Memphis among blood donors, sickle cell disease (SCD) patients and Amazonian Indians. Methods: A total of 427 blood samples were collected and separated into three groups: 206 unrelated blood donors, 90 patients with SCD and 131 Amazonian Indians. We performed DI*A/B, normal Band 3 and Band 3 Memphis genotyping, using the Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR-RFLP). Results: The frequency of the DI*A/DI*A genotype was 0.5% in blood donors and it was not found in other groups. The frequency of the DI*A/DI*B was higher in Amazonian Indians (33.6%) and the frequency of the DI*B/DI*B was highest in blood donors (92.2%). All 105 individuals tested were positive for the presence of normal Band 3 and of these individuals, only 5/105 (4.8%) presented the Band 3 Memphis mutation. Conclusion: We observed a higher frequency of the DI*B allele in blood donors and a low frequency of the DI*A/DI*A genotype in all groups studied. The Band 3 Memphis was found in a higher frequency in the blood donor group. Our findings highlight the importance of analyzing different population groups to gain a better understanding of the genetic association of blood group antigens.

Metabolomic profile in patients with primary warm autoimmune haemolytic anaemia.

Autoimmune haemolytic anaemia (AIHA) is a rare clinical condition with immunoglobulin fixation on the surface of erythrocytes, with or without complement activation. The pathophysiology of AIHA is complex and multifactorial, presenting functional abnormalities of T and B lymphocytes that generate an imbalance between lymphocyte activation, immunotolerance and cytokine production that culminates in autoimmune haemolysis. In AIHA, further laboratory data are needed to predict relapse and refractoriness of therapy, and thus, prevent adverse side-effects and treatment-induced toxicity. The metabolomic profile of AIHA has not yet been described. Our group developed a cross-sectional study with follow-up to assess the metabolomic profile in these patients, as well as to compare the metabolites found depending on the activity and intensity of haemolysis. We analysed the plasma of 26 patients with primary warm AIHA compared to 150 healthy individuals by mass spectrometry. Of the 95 metabolites found in the patients with AIHA, four acylcarnitines, two phosphatidylcholines (PC), asymmetric dimethylarginine (ADMA) and three sphingomyelins were significantly increased. There was an increase in PC, spermine and spermidine in the AIHA group with haemolytic activity. The PC ae 34:3/PC ae 40:2 ratio, seen only in the 12-month relapse group, was a predictor of relapse with 81% specificity and 100% sensitivity. Increased sphingomyelin, ADMA, PC and polyamines in patients with warm AIHA can interfere in autoantigen and autoimmune recognition mechanisms in a number of ways (deficient action of regulatory T lymphocytes on erythrocyte recognition as self, negative regulation of macrophage nuclear factor kappa beta activity, perpetuation of effector T lymphocyte and antibody production against erythrocyte antigens). The presence of PC ae 34:3/PC ae 40:2 ratio as a relapse predictor can help in identifying cases that require more frequent follow-up or early second-line therapies.

Frequency of the DI*A, DI*B and Band 3 Memphis polymorphism among distinct groups in Brazil.

INTRODUCTION: The Band 3 is a red blood cell protein that carries the Dia and Dib antigens from the Diego blood system. The SLC4A1 gene encodes Band 3; Band 3 Memphis is a polymorphism of normal Band 3 and has two variants, but only the variant II carries the Dia antigen. OBJECTIVES: Describe the frequencies of the DI*A and DI*B alleles and the Band 3 Memphis among blood donors, sickle cell disease (SCD) patients and Amazonian Indians. METHODS: A total of 427 blood samples were collected and separated into three groups: 206 unrelated blood donors, 90 patients with SCD and 131 Amazonian Indians. We performed DI*A/B, normal Band 3 and Band 3 Memphis genotyping, using the Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR-RFLP). RESULTS: The frequency of the DI*A/DI*A genotype was 0.5% in blood donors and it was not found in other groups. The frequency of the DI*A/DI*B was higher in Amazonian Indians (33.6%) and the frequency of the DI*B/DI*B was highest in blood donors (92.2%). All 105 individuals tested were positive for the presence of normal Band 3 and of these individuals, only 5/105 (4.8%) presented the Band 3 Memphis mutation. CONCLUSION: We observed a higher frequency of the DI*B allele in blood donors and a low frequency of the DI*A/DI*A genotype in all groups studied. The Band 3 Memphis was found in a higher frequency in the blood donor group. Our findings highlight the importance of analyzing different population groups to gain a better understanding of the genetic association of blood group antigens.

Incidentes transfusionais imediatos notificados em crianças e adolescentes / Reacciones transfusionales inmediatas notificadas en niños y adolescentes / Immediate transfusion incidents reported in children and adolescents

Resumo Objetivo Analisar o perfil dos incidentes transfusionais imediatos notificados em crianças e adolescentes internados em hospital geral de alta complexidade. Métodos Estudo documental retrospectivo, com análise de 287 Fichas de Notificação, reportando reações transfusionais em crianças e adolescentes de zero a 18 anos de idade, ocorridas no período de janeiro de 2007 e maio de 2021, em um serviço de hemovigilância de um Hospital Universitário, de caráter filantrópico, de alta complexidade, localizado na cidade de São Paulo. Resultados Das 287 fichas avaliadas, 42,5% das reações foram observadas em adolescentes (entre 12 e 18 anos), 83,6% ocorreram na primeira transfusão. Manifestações clínicas mais comuns foram lesões de pele e hipertermia. Cerca de 50% das reações ocorreram em pacientes com leucemia ou anemias e o hemocomponente associado foi o concentrado de hemácias. Incidentes mais comuns foram: reação febril não hemolítica e reações alérgicas, em sua maioria leves e moderadas. Outras reações foram 9,8% moderadas/graves. Conclusão O estudo favoreceu maior conhecimento sobre os incidentes transfusionais ocorridos em crianças e adolescentes e traz contribuições para reforçar a segurança do paciente e dos serviços de hemoterapia pediátrica.

Resumen Objetivo Analizar el perfil de las reacciones transfusionales inmediatas notificadas en niños y adolescentes internados en un hospital de alta complejidad. Métodos Estudio documental retrospectivo, con análisis de 287 Fichas de Notificación, donde se reportaron reacciones transfusionales en niños y adolescentes de cero a 18 años, ocurridas en el período de enero de 2007 a mayo de 2021, en un servicio de hemovigilancia de un hospital universitario de carácter filantrópico, de alta complejidad, ubicado en la ciudad de São Paulo. Resultados De las 287 fichas analizadas, el 42,5 % de las reacciones fue observada en adolescentes (entre 12 y 18 años) y el 83,6 % sucedió en la primera transfusión. Las manifestaciones clínicas más comunes fueron lesiones en la piel e hipertermia. Cerca del 50 % de las reacciones ocurrió en pacientes con leucemia o anemia, y el componente sanguíneo asociado fue el concentrado de eritrocitos. Los incidentes más comunes fueron: reacción febril no hemolítica y reacciones alérgicas, en su mayoría leves y moderadas. Otras reacciones fueron 9,8 % moderadas/graves. Conclusión El estudio ayudó a tener mayores conocimientos sobre los incidentes transfusionales ocurridos en niños y adolescentes y contribuye para reforzar la seguridad del paciente y de los servicios de hemoterapia pediátrica.

Abstract Objective To analyze the profile of immediate transfusion incidents reported in children and adolescents hospitalized in a high complexity general hospital. Methods This is a documentary and retrospective study that analyzed 287 notification records, reporting transfusion reactions in children and adolescents from zero to 18 years of age, occurred from January 2007 to May 2021, in a hemovigilance service of a high-complexity philanthropic university hospital in the city of São Paulo. Results Of the 287 records assessed, 42.5% of reactions were observed in adolescents (between 12 and 18 years), and 83.6% occurred in the first transfusion. Most common clinical manifestations were skin lesions and hyperthermia. About 50% of reactions occurred in patients with leukemia or anemia and the associated blood component was red blood cell concentrate. Most common incidents were nonhemolytic febrile reaction and mostly mild and moderate allergic reactions. Other reactions were 9.8% moderate/severe. Conclusion The study favored greater knowledge about transfusion incidents in children and adolescents and brings contributions to enhance patient safety and pediatric hemotherapy services.

HLA-DRB1 molecules and the presentation of anchor peptides from RhD, RhCE, and KEL proteins.

BACKGROUND: Antigens from the Rh and Kell systems are recognized as the most immunogenic in clinical practice. This study evaluated the possible molecular mechanisms involved in the interaction of antigenic peptides with the DRB1 molecules, which help to explain the high frequency of anti-K and association of D + C antibodies in transfusion and incompatible pregnancy. STUDY DESIGN AND METHODS: We included 201 patients with antibodies against antigens from the Rh and Kell systems and compare them with 174,015 controls. HLA-DRB1 genotyping and in silico analysis were performed. The NetMHCIIpan software was used to identify RhD-, RhCE-, and KEL-derived anchor peptides that bind to DRB1 molecules. RESULTS: HLA-DRB1*15 is associated with an increased risk of D, C, E, and K alloimmunization, while the HLA-DRB1*01 and *12 alleles are overrepresented in patients with anti-C and anti-D, respectively. In silico analysis showed that three polymorphic points (60I, 68S, and 103S) common to C and D antigens can be presented by several DRB1 molecules, including DRB1*15:01. The DRB1*09:01 molecule, although not showing statistical significance, was able to interact strongly with almost all five anchor peptides from the sequence containing the polymorphic determinants of E antigen, except 217-WMFWPSVNS-225. CONCLUSION: The DRB1*15 molecule has specific physicochemical characteristics in residues 11P and 13R in the P4 pocket that can favor the response to various antigenic peptides. Anti-K alloimmunization is unrestricted for interaction with specific DRB1 molecules, which suggests that almost all individuals in our population have DRB1 molecules capable of binding to KEL-derived anchor peptides and produce anti-K when stimulated.

Serologic and molecular studies to identify neonatal alloimmune neutropenia in a cohort of 10,000 neonates.

Neonatal alloimmune neutropenia (NAIN) is caused by maternal alloimmunisation to fetal human neutrophil antigens (HNAs). This study investigated maternal HNA/HLA alloantibodies involved with NAIN and identified the frequency of NAIN in Brazilian neonates. Neonatal neutropenia (neutrophil count < 1.5 × 109 /L) was investigated in samples from 10,000 unselected neonates, resulting in 88 neutropenic newborns (NBs) and their 83 mothers. Genotyping was performed by PCR-SSP (HNA-1/-4) and PCR-RFLP (HNA-3/-5). Serologic studies were performed by GAT (granulocyte agglutination test), Flow-WIFT (white blood cells immunofluorescence test) and LABScreen-Multi-HNA-Kit (OneLambda®) (LSM). Neonatal neutropenia was identified in 88/10,000 (0·9%) NBs. Genotyping revealed 60·2% maternal-fetal HNA incompatibilities (31·8% for HNA-1; 14·8% for HNA-3; 15·9% for HNA-4; 21·6% for HNA-5). Serologic studies revealed 37·3% of mothers with positive results with at least one technique. The detected anti-HNA specificities were confirmed in eight positive cases related to HNA-1/-3 systems. In cases with maternal-fetal HNA-4/-5 incompatibility, no specific neutrophil alloantibodies were found but anti-HLA I/II were present. Anti-HNA-2 was not identified. This is a large Brazilian study which involved the investigation of antibodies against all five HNA systems in neutropenia cases and showed a frequency of NAIN in 8/10,000 neonates. Among the HNA antibodies identified, we highlight the anti-HNA-1d and anti-HNA-3b, antibodies unusual in alloimmunised women, and rarely related to NAIN cases.

Antibodies against human neutrophil antigens in non-transfused women with red blood cell alloimmunisation induced by pregnancy.

BACKGROUND: Alloantibodies against human neutrophil antigens (HNA) resulting from allogeneic exposure may be associated with transfusion-related acute lung injury and immune neutropenia. Understanding the risk factors for the formation of such antibodies could have a great impact on the adoption of measures to prevent potentially fatal transfusion reactions. The aim of the study was to determine the prevalence of anti-HNA alloantibodies in non-transfused pregnant women with and without red blood cell (RBC) alloantibodies. MATERIALS AND METHODS: HNA alloantibodies were investigated in blood samples from 147 pregnant women with RBC alloimmunisation induced by pregnancy as the only allogeneic stimulus (group 1). The control group (group 2) consisted of 563 women with at least one pregnancy without RBC alloimmunisation. Both groups were investigated for the presence and identity of HNA alloantibodies using granulocyte agglutination tests, white blood cell immunofluorescence testing, and the bead-based LABScreen Multi Kit. Genotyping was performed to confirm the specificity of the HNA alloantibodies. RESULTS: Group 1 women had a statistically higher number of HNA alloantibodies compared to group 2 women (9/147 [6.1%] vs 9/563 [1.6%]; p=0.005, OR=4.01; 95% CI 1.5-10.3). Considering only multiparous women, there was a higher statistical significance for the difference in the presence of HNA alloantibodies between the two groups (7/82 [8.5%] vs 9/493 [1.8%]; p=0.002, OR=5.02; 95% CI 1.8-13.9). DISCUSSION: Our data show that RBC alloimmunisation is significantly associated with the development of anti-HNA alloantibodies, corroborating the hypothesis that some individuals are better immune responders and react strongly to allogeneic exposure. The presence of RBC alloantibodies can, therefore, facilitate the identification of individuals with a higher risk of alloimmunisation to antigens from other cells, also acting as a tool to avoid potentially fatal transfusion reactions.

RHD and RHCE molecular analysis in weak D blood donors and in patients with Rh antibodies against their own corresponding Rh antigen.

BACKGROUND: The Rh system is the largest and most polymorphic blood group system. The existence of a large number of RH alleles results in variant phenotypes that often complicate blood donor phenotyping and the distinction between auto- and allo-antibodies in recipients who have anti-Rh antibodies in the presence of their own corresponding Rh antigen. Knowledge of these variants is necessary in order to make blood transfusion safer. MATERIALS AND METHODS: Samples from 48 blood donors with serological weak D and from 29 patients who had anti-Rh antibody in the presence of their own corresponding Rh antigen were evaluated molecularly for RHD and RHCE alleles using a blood-multiplex ligation-dependent probe amplification assay and Sanger sequencing. RESULTS: Rh variants were found in 45 of the 48 blood donors: 24/45 (53%) were weak D, 2/45 (4%) partial D and 19/45 (42%) were weak and partial D. The remaining three donors (6%) did not show a mutation in the RHD allele. Among the 29 patients, 13/29 had anti-e, of whom 4/13 had genotypes that predicted a partial e antigen; 11/29 had anti-D, with 6/11 being identified as partial D; 2/29 had anti-c, of whom 1/2 was predicted to express partial c antigen; 4/29 who had anti-E and 4/29 who had anti-C did not show mutations in RHCE*C or RHCE*E. DISCUSSION: It was possible to find individuals with clinically significant Rh phenotypes due to the weak reactivity of the D antigen, detected through serological tests in blood donors. In patients, when found with the anti-Rh antibody in the presence of the same Rh antigen, it is difficult to distinguish an auto-antibody from an allo-antibody by serological tests; in these cases, molecular methods (genotyping) can help us to determine whether there are changes in the RH alleles and to discover the nature of the antibody (allo or auto).

BLOOD CELLS PROFILE IN UMBILICAL CORD OF LATE PRETERM AND TERM NEWBORNS.

OBJECTIVE: To describe the hematological profile in cord blood of late preterm and term newborns and compare blood indices according to sex, weight for gestational age and type of delivery. METHODS: Cross-sectional study with late preterm and term newborns in a second-level maternity. Multiple gestation, chorioamnionitis, maternal or fetal hemorrhage, suspected congenital infection, 5-minute Apgar <6, congenital malformations, and Rh hemolytic disease were excluded. Percentiles 3, 5,10, 25, 50, 75, 90, 95 and 97 of blood indices were calculated for both groups. RESULTS: 2,662 newborns were included in the sample, 51.1% males, 7.3% late preterms, 7.8% small for gestational age (SGA) and 81.2% adequate for gestational age (AGA). Mean gestational age was 35.6±1.9 and 39.3±1.0 weeks, respectively, for premature and term neonates. The erythrocytes indices and white blood cells increased from 34-36.9 to 37-41.9 weeks. Basophils and platelets remained constant during gestation. Premature neonates presented lower values ​​of all blood cells, except for lymphocytes and eosinophils. SGA neonates presented higher values ​​of hemoglobin, hematocrit and lower values of leukocytes, neutrophils, bands, segmented, eosinophils, monocytes and platelets. Male neonates presented similar values ​​of erythrocytes and hemoglobin and lower leukocytes, neutrophils, segmented and platelets. Neonates delivered by C-section had lower values ​​of red blood cells and platelets. Chronic or gestational hypertension induced lower number of platelets. CONCLUSIONS: Blood cells increased during gestation, except for platelets and basophils. SGA neonates had higher hemoglobin and hematocrit values and lower leukocytes. Number of platelets was smaller in male SGAs, born by C-section and whose mothers had hypertension.

Blood cells profile in umbilical cord of late preterm and term newborns / Perfil de hemograma em sangue de cordão umbilical de recém-nascidos pré-termo tardio e a termo

ABSTRACT Objective: To describe the hematological profile in cord blood of late preterm and term newborns and compare blood indices according to sex, weight for gestational age and type of delivery. Methods: Cross-sectional study with late preterm and term newborns in a second-level maternity. Multiple gestation, chorioamnionitis, maternal or fetal hemorrhage, suspected congenital infection, 5-minute Apgar <6, congenital malformations, and Rh hemolytic disease were excluded. Percentiles 3, 5,10, 25, 50, 75, 90, 95 and 97 of blood indices were calculated for both groups. Results: 2,662 newborns were included in the sample, 51.1% males, 7.3% late preterms, 7.8% small for gestational age (SGA) and 81.2% adequate for gestational age (AGA). Mean gestational age was 35.6±1.9 and 39.3±1.0 weeks, respectively, for premature and term neonates. The erythrocytes indices and white blood cells increased from 34-36.9 to 37-41.9 weeks. Basophils and platelets remained constant during gestation. Premature neonates presented lower values ​​of all blood cells, except for lymphocytes and eosinophils. SGA neonates presented higher values ​​of hemoglobin, hematocrit and lower values of leukocytes, neutrophils, bands, segmented, eosinophils, monocytes and platelets. Male neonates presented similar values ​​of erythrocytes and hemoglobin and lower leukocytes, neutrophils, segmented and platelets. Neonates delivered by C-section had lower values ​​of red blood cells and platelets. Chronic or gestational hypertension induced lower number of platelets. Conclusions: Blood cells increased during gestation, except for platelets and basophils. SGA neonates had higher hemoglobin and hematocrit values and lower leukocytes. Number of platelets was smaller in male SGAs, born by C-section and whose mothers had hypertension.

RESUMO Objetivo: Descrever o perfil hematológico em sangue de cordão de recém-nascidos pré-termo tardio e a termo e comparar parâmetros hematimétricos segundo sexo, adequação peso idade gestacional e tipo de parto. Métodos: Estudo transversal com recém-nascidos pré-termo tardio e a termo, em maternidade de nível secundário. Excluíram-se gestação múltipla, corioamnionite, hemorragia materna ou fetal, suspeita de infecção congênita, Apgar no 5o minuto <6, malformações congênitas e doença hemolítica Rh. Calcularam-se os percentis 3, 5, 10, 25, 50, 75, 90, 95 e 97 dos parâmetros hematológicos. Resultados: Incluíram-se 2.662 recém-nascidos, 51,1% do sexo masculino, 7,3% prematuros tardios, 7,8% pequenos para a idade gestacional e 81,2% adequados. A idade gestacional foi 35,6±1,9 e 39,3±1,0 semanas, respectivamente, nos prematuros e termos. As séries vermelha e branca aumentaram de 34-36,9 para 37-41,9 semanas, exceto basófilos e plaquetas, que permaneceram constantes. Os prematuros apresentaram menores médias nas séries vermelha, plaquetária e branca, com exceção de linfócitos e eosinófilos. Recém-nascidos pequenos para a idade gestacional apresentaram maiores valores de hemoglobina e hematócrito e menores de leucócitos, neutrófilos, bastonetes segmentados, eosinófilos, monócitos e plaquetas. Recém-nascidos masculinos apresentaram taxas semelhantes de hemoglobina e hematócrito e menores de leucócitos, neutrófilos, segmentados e plaquetas. Na cesárea, as células vermelhas e as plaquetas foram menores que no parto vaginal. O número de plaquetas foi menor na hipertensão crônica ou gestacional. Conclusões: As células sanguíneas aumentaram durante a gestação, exceto plaquetas e basófilos. Recém-nascidos pequenos para a idade gestacional apresentaram maiores taxas de hemoglobina e hematócrito e menores de células brancas. O número de plaquetas foi menor no recém-nascido pequeno para a idade gestacional, masculino, nascido por cesárea e de mãe hipertensa.

Molecular matching for patients with haematological diseases expressing altered RHD-RHCE genotypes.

BACKGROUND AND OBJECTIVES: The high homology and the inverted orientation of RHD and RHCE may give rise to non-functional and aberrant RH alleles. RH genotyping is used to screen RH matched donors to African descent patients. This study aimed to define a strategy for testing RHD and RHCE variants in blood donors to provide compatible units for transfusion of patients with haematological diseases. MATERIALS AND METHODS: Samples from 132 patients [101 Sickle cell disease (SCD), 14 myelodysplastic syndrome (MDS), 17 acute myelogenous leukaemia (AML)] and 198 Brazilian donors were studied. Major blood group alleles, RHD, RHCE alleles and RHD zygosity were determined by the blood-MLPA assay. Sequencing was performed to determine RHD and RHCE variant subtypes. A match was an RH genotype that did not encode Rh antigens absent in the patient, along with matching for ABO, MNS, KEL, FY, JK and DI antigens. RESULTS: Overall, 7·6% of blood donors and 17.4% of patients presented RH genotypes that predict expression of partial Rh antigens or lack of high prevalence Rh antigens. From 23 patients with clinically relevant RH genotypes, 15 had available matched donors. CONCLUSION: We report the presence of clinically relevant RH genotypes in SCD and in non-SCD patients. In our admixed population, many patients carry variant RHCE alleles in heterozygosity with normal RHCE alleles. Thus, our results suggest that donors could be selected based on the normal RH allele.

Frequência das reações transfusionais imediatas ocorridas em hospital de ensino em São Paulo, Brasil / The frequency of immediate transfusions reactions occurred in a teaching hospital from Sao Paulo, Brazil / La frecuencia de las Reacciones transfusionales inmediatas ocurridas en hospital de enseñanza en Sao Paulo, Brasil

Objetivo: descrever a ocorrência das reações transfusionais imediatas recebidas na Gerência de Risco do Hospital São Paulo. Metodologia: foi realizado um estudo transversal retrospectivo, com analises das fichas de notificação de Reações Transfusionais no período de maio de 2002 a dezembro de 2016 que foram inseridas no SNH. Resultados: foram analisadas 1548 fichas de RT, em sua quase totalidade reações imediatas, associadas ao Concentrado de Hemácias (72,5%). A mais comum foi a Reação Febril Não Hemolítica leve, sendo entre as graves e moderadas a Reação Alérgica. Os sinais e sintomas mais notificados foram a hipertermia, a sudorese, calafrios e lesões em derme. Não foram observadas diferenças entre sexo e idade, 90,8% das Reações Transfusionais ocorreram em pacientes com Fator Rh(+). Conclusão: o estudo permitiu uma melhor avalição e compreensão das reações transfusionais o que permite maior qualidade no ciclo do sangue e uma maior segurança dos pacientes submetidos a terapia transfusional.

Objective: to describe the occurrence of the immediate transfusion reactions received at the Risk Management of Hospital São Paulo. Methodology: a cross-sectional retrospective study was carried out, with analyzes of the ATR (Adverse Transfusional Reactions) in the period from May 2002 to December 2016 which were inserted in the National Haemovigilance System (SNH). Results: it were analyzed 1548 ATR records, most of which were immediate reactions, associated with the Red blood cells Concentrate (72,5%). The most common slight reaction was FNHTR (Febrile non-hemolytic transfusion reaction), and being between severe and moderate reactions, The Allergic reaction. The most commonly reported signs and symptoms were hyperthermia, sweating, chills and skins lesions. No differences were observed between gender and age, 90.8% of ATR occurred in patients with RhD positive blood group. Conclusion: the study allowed a better evaluation and understanding of the transfusion reactions which allowed higher quality in the blood cycle and increased safety of patients undergoing transfusion therapy.

Objetivo: describir la ocurrencia de las reacciones transfusionales inmediatas recibidas en la Gestión de Riesgo del Hospital São Paulo. Metodología: se realizó un estudio transversal retrospectivo, con análisis de las fichas de notificación de Reacciones Transfusionales en el período de mayo de 2002 a diciembre de 2016 que fueron introducidas en el SNH. Resultados: se analizaron 1548 fichas de RT inmediatas, asociadas al Concentrado de Hemácias (72,5%). La más común fue la Reacción Febril no Hemolítica leve, siendo la reacción alérgica entre las graves y moderadas. Los signos y síntomas más notificados fueron la fiebre, la sudoración, escalofríos y lesiones en dermis. No se observaron diferencias entre sexo y edad, el 90,8% de las Reacciones Transfusionales ocurrieron en pacientes con Factor Rh(+). Conclusión: el estudio permitió una mejor evaluación y comprensión de las reacciones transfusionales, lo que permite mayor calidad en el ciclo de la sangre y una mayor seguridad de los pacientes sometidos a terapia transfusional.

Molecular study of Cw /Cx antigens and frequency of Rh phenotypes in southeast Brazilian blood donors.

BACKGROUND: The Cw (RH:8), Cx (RH:9), and MAR (RH:51) antigens are encoded by alleles at the Cc locus of the Rh system, where Cw and Cx are considered low-frequency antigens and antithetical to the high-frequency antigen MAR. The frequency of Cw (RH:8) is approximately 2% in Caucasians, 1% in Black people, 4% in Finns, and 9% in Latvians. The aim of this study was to determine the frequency of RhD+ phenotypes in a population of southeast Brazilian blood donors and to perform a molecular study to distinguish the RHCE*Ce.08.01 and RHCE*Ce.09 alleles, responsible for the Cw and Cx expressions, respectively. METHODS: We investigated 11,536 RhD+ Brazilian blood donors. All samples were phenotyped for D, C, c, E, e, and Cw . In the Cw + samples, a molecular analysis was performed to detect the nucleotide substitutions A122G and G106A, which determine the Cw and Cx antigens, respectively. RESULTS: Cw antigen was found in 110 (0.95%) samples in the following phenotypes: DCw e/dCw e (72/0.62%), DCw e/DCw e (30/0.26%), and DCw e/DCw E (8/0.07%). Among 110 Cw + samples, 108 showed the A122G nucleotide substitution associated with RHCE*Ce.08.01 allele and 2 samples the G106A substitution associated with the RHCE*Ce.09.01 allele. CONCLUSION: This study showed the prevalence of the RhD+ phenotype in the Brazilian population, and that through the molecular study, it was possible to differentiate the RHCE*Ce.08.01 and RHCE*Ce.09.01 alleles. The phenotype frequency was similar from Black people (1%) and different from Caucasians, Finns, and Latvians.

Antibodies to human neutrophil antigen HNA-3b implicated in cases of neonatal alloimmune neutropenia.

BACKGROUND: Neonatal alloimmune neutropenia results from maternal alloimmunization to human neutrophil antigens. The alloantibodies involved in neonatal alloimmune neutropenia are against human neutrophil antigens HNA-1a, HNA-1b, HNA-1c, HNA-1d, HNA-2, HNA-3a, HNA-4a, HNA-4b, and HNA-5a; however, to date, antibodies specific to HNA-3b have not been reported. STUDY DESIGN AND METHODS: Blood samples from 10,000 unselected neonates were analyzed, resulting in the selection of 88 neutropenic newborns (neutrophil count <1.5 × 109 /L) from 83 mothers (three pairs of twins and one triplet). HNA-3 genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism to identify the cases of maternal-fetal HNA-3 incompatibility. Serologic studies for detecting maternal HNA-3 alloantibodies were performed with the granulocyte agglutination test, the white blood cell immunofluorescence test, and a LABScreen Multi-HNA Kit. RESULTS: Genotyping studies identified 13 of 88 (14.8%) instances of maternal-fetal HNA-3 incompatibility, with all mothers typed as HNA-3a/a and neonates typed as HNA-3a/b. Serologic studies revealed that five of 13 (38.5%) mothers carried anti-HNA-3b plus human leukocyte antigen antibodies and that three of 13 (23.1%) mothers had anti-HNA-3b without human leukocyte antigen antibodies. CONCLUSION: Here, we report the first three cases of neonatal alloimmune neutropenia associated with HNA-3b antibodies resulting in a neonatal alloimmune neutropenia incidence of one in 3333 live births.

Molecular Basis of KELnull Phenotype in Brazilians.

BACKGROUND: KELnull (K0) persons can produce clinically significant anti-KEL5 antibody after transfusion and/or pregnancy, requiring K0 blood transfusion when indicated. 37 K0 alleles have been reported in studies over different populations, but none in Amerindian-Caucasian descendants from South America. The aim of this study was to identify the molecular basis of K0 phenotype in Brazilians. METHODS: We investigated three K0 samples from different Brazilian blood banks (Recife, Manaus, and Vila Velha) in women with anti-KEL5. KEL antigen typing was performed by serologic techniques, and the K0 status was confirmed by flow cytometry. PCR-RFLP and DNA sequencing of the KEL coding and exon-intron regions were also performed. RESULTS: RBCs of the 3 patients were phenotyped as KEL:-1,-2,-3,-4,-7. The 3 patients had the same KEL*02/02 genotype and were negative for KEL*02.03 and KEL*02.06 alleles. The Recife K0 patient was homozygous for IVS16 + 1g>a mutation (KEL*02N.31 allele). The flow cytometry with anti-KEL1, anti-KEL2, anti-KEL3, anti-KEL4, and anti-CD238 confirmed the K0 phenotype. In addition, we found the c.10423C>T mutation (KEL*02N.04 allele) in both the Manaus K0 and the Vila Velha K0 patients. CONCLUSION: This report represents the first study of K0 molecular basis performed in Amerindian-Caucasian descendants from South America.

HNA-3 gene frequencies in Brazilians and a new polymerase chain reaction-restriction fragment length polymorphism method for HNA-3a/3b genotyping.

BACKGROUND: HNA-3 antigens are the result of a rs2288904 single-nucleotide polymorphism (SNP) in the CTL2, and the HNA-3a and HNA-3b variants are encoded by a guanine and adenine at Nucleotide Position 461. Anti-HNA-3 are involved in severe transfusion-related acute lung injury reactions and in neonatal alloimmune neutropenia. Since the distribution of the HNA-3 system was unknown in South Americans, in this study we determined the frequency of the HNA-3 alleles in Brazilians. STUDY DESIGN AND METHODS: DNA of 500 blood donors, 120 Xikrin Amerindians, 74 Japanese individuals, and 124 African Brazilians were genotyped for rs2288904 by a polymerase chain reaction (PCR)-restriction fragment length polymorphism assay. The PCR product was digested with enzyme Taq(α) 1, specific to nucleotide guanine (HNA-3a). RESULTS: The results showed that the frequencies of the HNA-3a/HNA-3b alleles were 0.81/0.19 in blood donors, 1.00/0.00 in Amerindians, 0.63/0.37 in Japanese, and 0.85/0.15 in African Brazilians. All 81 individuals genotyped as HNA-3a/a did not present the SNP c.457T by molecular sequencing. CONCLUSION: The frequencies of HNA-3 genotypes in Brazilian blood donors is similar to that described in Caucasians; however, all Amerindians were HNA-3a/a, African Brazilians showed a lower frequency of HNA-3b/b, and Japanese had a higher prevalence of HNA-3b/b, suggesting that they may be at risk for developing anti-HNA-3a alloantibodies.

Reduction of exposure to blood donors in preterm infants submitted to red blood cell transfusions using pediatric satellite packs.

OBJECTIVE: In preterm newborn infants transfused with erythrocytes stored up to 28 days, to compare the reduction of blood donor exposure in two groups of infants classified according to birth weight. METHODS: A prospective study was conducted with preterm infants with birth weight <1000 g (Group 1) and 1000-1499 g (Group 2), born between April, 2008 and December, 2009. Neonates submitted to exchange transfusions, emergency erythrocyte transfusion, or those who died in the first 24 hours of life were excluded. Transfusions were indicated according to the local guideline using pediatric transfusion satellite bags. Demographic and clinical data, besides number of transfusions and donors were assessed. . Logistic regression analysis was performed to determine factors associated with multiple transfusions. RESULTS: 30 and 48 neonates were included in Groups 1 and 2, respectively. The percentage of newborns with more than one erythrocyte transfusion (90 versus 11%), the median number of transfusions (3 versus 1) and the median of blood donors (2 versus 1) were higher in Group 1 (p<0.001), compared to Group 2. Among those with multiple transfusions, 14 (82%) and one (50%) presented 50% reduction in the number of blood donors, respectively in Groups 1 and 2. Factors associated with multiple transfusions were: birth weight <1000 g (OR 11.91; 95%CI 2.14-66.27) and presence of arterial umbilical catheter (OR 8.59; 95%CI 1.94-38.13), adjusted for confounders. CONCLUSIONS: The efficacy of pediatrics satellites bags on blood donor reduction was higher in preterm infants with birth weight <1000 g.

Reduction of exposure to blood donors in preterm infants submitted to red blood cell transfusions using pediatric satellite packs / Reduccion de la exposicion a donantes de sangre en prematuros sometidos a transfusiones de hematies con uso de bolsas de transferencia pediatrica / Reducao da exposicao a doadores de sangue em prematuros submetidos a transfusoes de hemaciascom uso de bolsas de transferencia pediatricas

OBJECTIVE: In preterm newborn infants transfused with erythrocytes stored up to 28 days, to compare the reduction of blood donor exposure in two groups of infants classified according to birth weight. METHODS: A prospective study was conducted with preterm infants with birth weight <1000g (Group 1) and 1000-1499g (Group 2), born between April, 2008 and December, 2009. Neonates submitted to exchange transfusions, emergency erythrocyte transfusion, or those who died in the first 24 hours of life were excluded. Transfusions were indicated according to the local guideline using pediatric transfusion satellite bags. Demographic and clinical data, besides number of transfusions and donors were assessed. . Logistic regression analysis was performed to determine factors associated with multiple transfusions. RESULTS: 30 and 48 neonates were included in Groups 1 and 2, respectively. The percentage of newborns with more than one erythrocyte transfusion (90 versus 11%), the median number of transfusions (3 versus 1) and the median of blood donors (2 versus 1) were higher in Group 1 (p<0.001), compared to Group 2. Among those with multiple transfusions, 14 (82%) and one (50%) presented 50% reduction in the number of blood donors, respectively in Groups 1 and 2. Factors associated with multiple transfusions were: birth weight <1000g (OR 11.91; 95%CI 2.14-66.27) and presence of arterial umbilical catheter (OR 8.59; 95%CI 1.94-38.13), adjusted for confounders. CONCLUSIONS: The efficacy of pediatrics satellites bags on blood donor reduction was higher in preterm infants with birth weight <1000g. .

OBJETIVO En prematuros transfundidos con hematíes preservados por hasta 28 días, comparar la reducción de exposición a donantes en dos grupos de pacientes, según el peso al nacer. MÉTODO Se trata de un estudio prospectivo con prematuros con peso al nacer <1000g (Grupo 1) y de 1000-1499g (Grupo 2), nacidos entre abr/2008 a dic/2009. Se excluyeron recién-nacidos sometidos a exsanguineotransfusión, transfusión de emergencia u óbito antes de 24 horas de vida. Las transfusiones fueron indicadas según la rutina del servicio, utilizando bolsas de transferencia pediátrica. Se analizaron datos demográficos, clínicos y número de transfusiones y donantes. Variables categóricas fueron comparadas por la prueba de chi-cuadrado y numéricas por la prueba t o Mann-Whitney. Se utilizó regresión logística para análisis de factores asociados a las múltiples transfusiones. RESULTADOS: Se incluyeron 30 prematuros en el Grupo 1 y 48 en el Grupo 2. El porcentaje de prematuros que recibió más de una transfusión de hematíes (89,5 versus 10,5%), la mediana del número de transfusiones (3 versus 1) y la mediana de donantes (2 versus 1) fue mayor en el Grupo 1, comparado al Grupo 2 (p<0,001). Entre aquellos con transfusiones múltiples, 14 (82,4%) y 1 (50,0%) prematuros presentaron reducción de 50% de donantes respectivamente en los Grupos 1 y 2. Los factores asociados a múltiples transfusiones fueron peso al nacer <1000g (OR 11,91; IC95% 2,14-66,27) y presencia de catéter arterial umbilical (8,59; 1,94-38,13), controlados para variables de confusión. CONCLUSIONES La eficacia de las bolsas de transferencia pediátricas para reducir la exposición a donantes de sangre fue mayor en prematuros con peso al nacer <1000g. .

OBJETIVO Em prematuros transfundidos com hemácias preservadas por até 28 dias, comparar a redução de exposição a doadores em dois grupos de pacientes, de acordo com o peso ao nascer. MÉTODOS Estudo prospectivo de prematuros com peso ao nascer <1000g (Grupo 1) e de 1000-1499g (Grupo 2), nascidos entre abril de 2008 e dezembro de 2009. Excluíram-se os recém-nascidos submetidos a exsanguineotransfusão, transfusão de emergência ou óbito antes de 24 horas de vida. As transfusões foram indicadas conforme rotina do serviço, utilizando-se bolsas de transferência pediátricas. Analisaram-se dados demográficos, clínicos e número de transfusões e doadores. Utilizou-se regressão logística para análise de fatores associados às múltiplas transfusões. RESULTADOS Incluíram-se 30 prematuros no Grupo 1 e 48 no Grupo 2. A porcentagem de prematuros que receberam mais de uma transfusão de hemácias (90 versus 11%), a mediana do número de transfusões (3 versus 1) e mediana de doadores (2 versus 1) foram maiores no Grupo 1, comparado ao Grupo 2 (p<0,001). Entre aqueles com transfusões múltiplas, 14 (82%) e um (50%) prematuros apresentaram redução de 50% de doadores respectivamente nos Grupos 1 e 2. Os fatores associados a múltiplas transfusões foram peso ao nascer <1000g (OR 11,91; IC95% 2,14-66,27) e presença de cateter arterial umbilical (OR 8,59; IC95% 1,94-38,13), controlados para variáveis de confusão. CONCLUSÕES A eficácia das bolsas de transferência pediátricas para reduzir a exposição a doadores de sangue foi maior em prematuros com peso ao nascer <1000g. .