Zika virus RNA detection in blood donors in São Paulo, Brazil

ABSTRACT Introduction: The Zika Virus (ZIKV) is a single-stranded RNA genome virus, belonging to the family Flaviviridae, genus Flavivirus. Outbreaks around the world have demonstrated that the presence of asymptomatic viremic blood donors provides an increase in the risk of transfusion transmission (TT) and nucleic acid test (NAT) screening has been proposed to ensure the blood safety. This study implemented an "in-house" method to detect ZIKV RNA in blood sample donations. Methods: Primary plasma tubes are submitted to nucleic acid extraction on an automated platform. After extraction, the NAT set-up is performed in the robotic pipettor, in which an amplification mixture containing primers and probes for ZIKV and Polio vaccine virus (PV) are added in duplex as an internal control. The real-time polymerase chain reaction is then performed in a thermocycler, using the protocol established by the supplier. Results: From May 2016 to May 2018, 3,369 samples were collected from 3,221 blood donors (confidence coefficient 95%), of which 31 were considered false positive (0.92%), as they did not confirm initial reactivity when repeated in duplicates and 14 (0.42%) had their results invalid due to repeat failure in the internal control, 4 (0.12%), due to insufficient sample volume and 2 (0.05%), due to automatic pipettor failures. No Zika RNA reactive sample was identified. Conclusion: The test showed feasible to be incorporated into the blood screening routine. Our data do not indicate the need to screen for ZIKV RNA in São Paulo during the evaluated period. However, a generic NAT system covering a group of flaviviruses which are circulating in the region, such as DENV and YFV, among others, could be a useful tool.

The relevance of a bank with genotyped platelets donors

ABSTRACT Objective: Describe the clinical and laboratory characteristics and the transfusion strategy of patients at Hospital Israelita Albert Einstein with platelet refractoriness and identify their etiological characteristics. Standardize the platelet immunofluorescence technique by flow cytometry as a test for platelet compatibility in immune platelet refractoriness in transfusion support. Methods: Review of medical records of refractory platelet patients followed at HIAE from January 2011 to May 2017. Clinical-demographic data, laboratory data and identification of the use of compatible genotyped platelets for patients in need of transfusion therapy were collected. The analyzed patients were classified according to the etiology of their platelet refractoriness. To standardize the FC-PIFT technique, blood group O platelets were incubated with serum from blood group AB donors and anti-IgG monoclonal antibody to determine the negative control. In order to verify the influence of the ABO system, monoclonal anti-IgG antibodies were incubated with blood group A or B platelets and with blood group O donor serum with isohemagglutinins below and above 1/64. Results: A total of 47 patients were evaluated, a 51% (24/47) preponderance of associated immune and non-immune factors (NIPR + IPR). The most common causes of NIPR + IPR were splenomegaly (54%) and the development of HLA antibodies (88%), consistent with the literature. For patients who required therapeutic transfusion, only a small portion received compatible genotyped platelets. Conclusion: Although 60% of patients could benefit from the therapeutic transfusion of genotyped platelets, only 10% were actually transfused with this type of blood component. This reaffirms the need for investments in a bank of genotyped platelet donors.

Platelet antibodies identification: comparison between two laboratory tests

ABSTRACT Introduction: Platelet antibody identification is indispensable for diagnosing the human platelet antigen (HPA) or human leukocyte antigen (HLA) immunization, mostly because it can restrict the compatibility and results of transfusions. Correct detection of these antibodies is of utmost importance for the diagnosis and treatment. Method: We present 16 platelet alloimmunization results, comparing two tests with different technologies: the MAIPA (monoclonal antibody immobilization of platelet antigens), as a reference technique, and a bead-based assay, the Pak-Lx. Results: Eleven samples (68.75%) showed agreement in both techniques. Two tests were false negatives in the Pak-Lx: a pan-reactivity in GPIIbIIIa and an anti-HPA-9b. On the other hand, the Pak-Lx was more sensitive to detect a decreasing anti-HPA-5b. The Pak-Lx found an anti-HPA-2b positive, but with a low median fluorescent intensity (MFI), suggesting a false-positive result. Moreover, in one case, the MAIPA was negative for a positive Pak-Lx HLA. Conclusion: Antibody platelet diagnosis can sometimes be challenging. The methods seemed similar, the Pak-Lx being faster and simpler than the MAIPA, and they can be complementary to solve clinical issues.

Matrix-assisted autologous chondrocyte transplantation for treatment of focal chondral lesions in the knee: the Hospital Israelita Albert Einstein experience

ABSTRACT Objective Phase 1 clinical trial to determine feasibility, safety, and efficacy of a new advanced cell therapy product for treatment of knee articular cartilage injuries. Methods Three participants with knee focal chondral lesions were included, with no signs of osteoarthritis. Chondrocytes were obtained through knee arthroscopy, cultured in collagen membrane for 3 weeks at the laboratory, subjected to tests to release the cell therapy product, and implanted. All patients underwent a specific 3-month rehabilitation protocol, followed by assessments using functional and imaging scales. The main outcome was the incidence of severe adverse events. Results Three participants were included and completed the 2-year follow-up. There was one severe adverse event, venous thrombosis of distal leg veins, which was no associated with therapy, was treated and left no sequelae. The clinical and radiological scales showed improvement in the three cases. Conclusion The preliminary results, obtained with the described methodology, allow concluding that this product of advanced cell therapy is safe and feasible. ReBEC platform registration number: RBR-6fgy76

Report of a clinical and laboratory management of cell therapy for knee cartilage in the face of mycoplasma contamination

ABSTRACT To describe a case of autologous chondrocyte implantation after cell culture contamination by Mycoplasma pneumoniae and the measures taken to successfully complete cell therapy in a patient with focal chondral lesion. A 45-year-old male patient, complaining of chronic pain on the knee and no history of trauma. He had a chondral lesion in the trochlear region of the femur and clinical tests compatible with pain in the anterior compartment of the knee. Conservative treatment failed to alleviate symptoms. Surgical treatment was indicated, but due to the size of the lesion, membrane-assisted autologous chondrocyte implantation was the technique of choice. Cartilage biopsies were collected from the intercondylar region of the distal femur. After isolation, chondrocytes were expanded ex vivo in a trained laboratory, for three weeks, and seeded onto a commercially available collagen membrane prior to implantation in the knee. Two days before surgery, a cell culture sample tested positive for Mycoplasma pneumoniae. The source of contamination was found to be autologous blood serum, extracted from the patient´s peripheral vein, and used to supplement the cell culture medium. After treating the patient with antibiotics, all procedures were repeated and the new final cell product, free from contaminants, was successfully implanted. We discuss the strategies available to deal with this situation, and describe the results of this particular case, which led to modifications in the autologous chondrocyte implant protocol.

Superiority of the triple combination of bortezomib, cyclophosphamide and dexamethasone versus cyclophosphamide, thalidomide and dexamethasone in patients with newly diagnosed multiple myeloma, eligible for transplantation

ABSTRACT Background: The treatment of multiple myeloma (MM) has evolved significantly in the past decade, and new drug combinations have improved the response rates and prolonged survival. Studies comparing different induction chemotherapy regimens have shown that triple combinations have better results than double combinations. However, comparisons among different triple combinations are rare in the literature. Methods: We retrospectively compared two triple combinations comprising bortezomib, cyclophosphamide and dexamethasone (VCD) versus thalidomide, cyclophosphamide and dexamethasone (CTD), and aimed at identifying which of the two combinations would yield better response rates following four induction cycles prior to hematopoietic cell transplantation in patients with untreated multiple myeloma. Results: We retrospectively reviewed the medical records of 311 patients from 24 different centers.The VCD regimen was used as induction therapy by 117 (37.6%) patients, whereas 194 (62.4%) patients received the CTD regimen. After four cycles of induction on an intention-to-treat basis, 54% of the patients in the VCD group achieved at least very good partial response versus 42.8% in the CTD group (p = 0.05). We observed no difference in neuropathy or thrombotic events rates among the two regimens. Conclusion: Our results corroborate the superiority of the triple combination regimes containing bortezomib over the triple combination with thalidomide as pre ASCT induction therapy in MM.

The rare holley antibody associated with a severe hemolytic transfusion reaction: the importance of this antibody identification to find a compatible blood unit / Raro anticorpo associado à reação transfusional hemolítica grave: a importância de sua identificação para encontrar uma unidade sanguínea compatível

ABSTRACT The correct identification of erythrocyte antibodies is fundamental for the searching for compatible blood and haemolytic transfusion reactions prevention. Antibodies against antigens of high prevalence are difficult to identify because of the rarity of their occurrence and unavailability of negative red cells for confirmation. We report a case of 46-years-old woman, diagnosed with hemoglobinopathy, and who had symptomatic fall in hemoglobin levels (5.3g/dL) after blood transfusion suggestive of transfusion reaction. The patient's blood type was O RhD-positive. Irregular antibody screening was positive and demonstrated a panreaction against all erythrocytes tested, but this result was not reactive with dithiothreitol. Using negative red cells for antigens of high prevalence of our inventory we could identify in the serum of the same erythrocytes an anti-Holley antibody associated with anti-E. Molecular analysis confirmed that the patient was negative for E and Holley antigens. The crossmath with compatible units confirmed the results. Holley is a high prevalence antigen of the Dombrock blood system whose negative phenotype is extremely rare in all populations and is associated with hemolytic transfusion reactions. This is an antibody that is difficult to identify because laboratories need to have experience in solving complex cases, and have available a large stock of rare sera and erythrocytes, as well other tools such as enzymes, thiol reagents and molecular tests. The correct identification of a rare antibody is initial and mandatory for searching of compatible donors, and to guarantee a satisfactory transfusional support.

RESUMO A correta identificação dos anticorpos eritrocitários é fundamental na busca de sangue compatível e na prevenção das reações transfusionais hemolíticas. Anticorpos contra antígenos de alta prevalência são de difícil identificação, devido à raridade de sua ocorrência e à indisponibilidade de hemácias negativas para sua confirmação. Apresentamos aqui o caso de uma paciente do sexo feminino, 46 anos, com diagnóstico de hemoglobinopatia, que apresentou queda sintomática dos níveis de hemoglobina (5,3g/dL) após transfusão sanguínea, sugestiva de reação transfusional. O tipo sanguíneo da paciente era O RhD-positivo. A pesquisa de anticorpos irregulares foi positiva, demonstrando panreação contra todos os eritrócitos testados, mas não reativo ao ditiotreitol. Utilizando hemácias selecionadas negativas para antígenos de alta prevalência do nosso inventário, foi possível identificar no soro da mesma um anticorpo anti-Holley associado a um anti-E. A análise molecular confirmou que a paciente era negativa para os antígenos E e Holley, e as provas de compatibilidade com unidades fenotipadas confirmaram os resultados. Holley é um antígeno de alta prevalência do sistema sanguíneo Dombrock, cujo fenótipo negativo é extremamente raro em todas as populações e está associado a reações transfusionais hemolíticas. Trata-se de anticorpo de difícil identificação, pois os laboratórios precisam ter experiência na resolução de casos complexos, grande estoque de soros e eritrócitos raros, além de outras ferramentas, como enzimas, reagentes tiol e testes moleculares. A identificação correta de um anticorpo raro é inicial e obrigatória para a busca de doadores compatíveis, garantindo um suporte transfusional satisfatório.

Allele and haplotype frequencies of human platelet and leukocyte antigens in platelet donors / Frequência alélica e haplotípica dos antígenos plaquetários e leucocitários humanos em doadores de plaquetas

ABSTRACT Objective To described the allele and haplotype frequencies of human leukocyte antigen genes at the -A, -B loci and human platelet antigen genes for human platelet antigen systems 1 to 9, 11 and 15 in blood. Methods We included 867 healthy unrelated volunteer donors who donated platelets between January 2011 and December 2014. Microarray genotyping was performed using a BeadChip microarray. Medium resolution typing of the human leukocyte antigen at loci A and B was carried out using sequence-specific oligonucleotide probe hybridization. We used multivariate analysis and our human leukocyte antigen population was compared to data from the United States national bone marrow donor program. Human platelet antigen results were compared to a literature review and data from around the world. Results Our human leukocyte antigen haplotype results were more similar to those of hispanics, followed by caucasians. Likewise, our human platelet antigen sample is more similar to those of Argentina, Rio Grande do Sul and Italy. Conclusion This was the first article that discusses human platelet antigen and human leukocyte antigen data together. Rare genotypes or antibody associations can make patient management difficult. A blood bank with genotyped donors allows for optimal transfusion and can contribute to better results. Our information can serve as basis for a database of platelet antigen polymorphisms.

RESUMO Objetivo Descrever as frequências alélicas e haplotípicas de genes dos antígenos leucocitários humanos nos loci -A,- B e dos antígenos plaquetários humanos para os sistemas HPA-1 a 9, 11 e 15. Métodos Foram incluídos 867 doadores voluntários, saudáveis, não relacionados, que doaram plaquetas por aférese entre janeiro de 2011 e dezembro de 2014. A genotipagem foi realizada usando microarray BeadChip. A tipificação de resolução intermediária dos antígenos leucocitários humanos loci A e B foi realizada por meio de hibridização com sonda para oligonucleotídeos por sequência específica. Utilizamos análises multivariadas e o antígeno leucocitário humano de nossa população foi comparado com a do programa nacional de doadores de medula óssea norte-americano. Já os resultados dos antígenos plaquetários humanos foram comparados à revisão da literatura e a dados de populações de outros países. Resultados Os resultados do haplótipo de antígenos leucocitários humanos são mais parecidos com os dos hispânicos, seguidos dos caucasianos. Igualmente, a amostra de antígenos plaquetários humanos foi mais semelhante às da Argentina, do Rio Grande do Sul e da Itália. Conclusão Este foi o primeiro artigo a discutir antígenos plaquetários e leucocitários humanos simultaneamente. Genótipos raros ou associações de anticorpos podem dificultar o manejo clínico do paciente. Um banco de sangue com doadores genotipados permite um melhor resultado e transfusão possíveis. Estas informações podem servir de base para um banco de dados sobre polimorfismos de antígenos plaquetários.

Transfusion management for patients taking an anti-CD38 monoclonal antibody

Abstract Introduction: Pre-transfusion tests, essential for the release of blood components, may be affected by drugs. Monoclonal antibodies represent a class of medications increasingly used in the clinical practice, with anti-CD38 monoclonal antibodies (daratumumab) being a promising resource in the treatment of refractory myeloma. This monoclonal antibody recognizes CD38 in myeloma cells and interferes with pre-transfusion tests by causing panreactivity in indirect antiglobulin tests thereby clinically masking alloantibodies. Dithiothreitol is a reagent that breaks disulfide bonds and effectively destroys antigenic sites for CD38 on red blood cells. This study reports the immunohematological findings of pre-transfusion tests of patients with multiple myeloma receiving daratumumab and on solutions to prevent the interference of this monoclonal antibody. Methods: Serum samples from five patients on anti-CD38 monoclonal antibody treatment were evaluated. Tests performed included ABO/RhD typing, indirect antiglobulin test, direct antiglobulin test and eluate test. A daily evaluation was performed to determine the shelf life of dithiothreitol-treated red blood cells when stored in Alsever's solution. Results: No interference in the ABO/RhD typing results was noted but in all samples, a panreactivity was observed in indirect antiglobulin tests. Regarding the direct antiglobulin test, two samples presented positive results but negative eluates. In all samples, treatment of reagent red blood cells with 0.2 M dithiothreitol offset interference by anti-CD38 monoclonal antibodies. Dithiothreitol-treated red blood cells stored in Alsever's solution were stable for up to 15 days. Conclusion: Treatment of reagent red blood cells with dithiothreitol can be efficient and accessible to offset the interference of the anti-CD38 drug in pre-transfusion tests. The number of costly serological workups can be reduced by having stored dithiothreitol red blood cells with this proving to be a useful reagent for investigating anti-CD38.

Determination of an unrelated donor pool size for human leukocyte antigen-matched platelets in Brazil

ABSTRACT Background: Successful transfusion of platelet refractory patients is a challenge. Many potential donors are needed to sustain human leukocyte antigen matched-platelet transfusion programs because of the different types of antigens and the constant needs of these patients. For a highly mixed population such as the Brazilian population, the pool size required to provide adequate platelet support is unknown. Methods: A mathematical model was created to estimate the appropriate size of an unrelated donor pool to provide human leukocyte antigen-compatible platelet support for a Brazilian population. A group of 154 hematologic human leukocyte antigen-typed patients was used as the potential patient population and a database of 65,500 human leukocyte antigen-typed bone marrow registered donors was used as the donor population. Platelet compatibility was based on the grading system of Duquesnoy. Results: Using the mathematical model, a pool containing 31,940, 1710 and 321 donors would be necessary to match more than 80% of the patients with at least five completely compatible (no cross-reactive group), partial compatible (one cross-reactive group) or less compatible (two cross-reactive group) donors, respectively. Conclusion: The phenotypic diversity of the Brazilian population has probably made it more difficulty to find completely compatible donors. However, this heterogeneity seems to have facilitated finding donors when cross-reactive groups are accepted as proposed by the grading system of Duquesnoy. The results of this study may help to establish unrelated human leukocyte antigen-compatible platelet transfusions, a procedure not routinely performed in most Brazilian transfusion services.

Successful management of neonatal alloimmune thrombocytopenia in the second pregnancy: a case report / Sucesso no manejo da trombocitopenia aloimune neonatal na segunda gestação: relato de caso

Neonatal alloimmune thrombocytopenia is a serious disease, in which the mother produces antibodies against fetal platelet antigens inherited from the father; it is still an underdiagnosed disease. This disease is considered the platelet counterpart of the RhD hemolytic disease of the fetus and newborn, yet in neonatal alloimmune thrombocytopenia the first child is affected with fetal and/or neonatal thrombocytopenia. There is a significant risk of intracranial hemorrhage and severe neurological impairment, with a tendency for earlier and more severe thrombocytopenia in subsequent pregnancies. This article reports a case of neonatal alloimmune thrombocytopenia in the second pregnancy affected and discusses diagnosis, management and the clinical importance of this disease.

A púrpura trombocitopênica neonatal aloimune é uma doença grave, na qual a mãe produz anticorpos contra antígenos plaquetários fetais herdados do pai, e é ainda subdiagnosticada na prática clínica. É considerada o equivalente plaquetário da doença hemolítica do recém-nascido, com a diferença que o primeiro filho é afetado, apresentando trombocitopenia fetal e/ou neonatal. Há risco significativo de hemorragia intracraniana e sequelas neurológicas graves, com tendência a trombocitopenia mais grave e mais precoce nas gestações subsequentes. Este artigo relata um caso de trombocitopenia aloimune neonatal na segunda gestação afetada e discute diagnóstico, manejo e importância clínica dessa doença na prática clínica.

One window-period donation in two years of individual donor-nucleic acid test screening for hepatitis B, hepatitis C and human immunodeficiency virus

Objective: To describe general data on nucleic acid/serology testing and report the first hepatitis B-nucleic acid testing yield case of an immunized donor in Brazil. Methods: A total of 24,441 donations collected in 2010 and 2011 were submitted to individual nucleic acid testing for hepatitis B, hepatitis C and human immunodeficiency virus using the TaqMan® MPX kit (Roche) on the Cobas s201 platform, in addition to routine screening for serological markers. Nucleic acid testing-reactive donations were further evaluated by real-time polymerase chain reaction using Cobas AmpliPrep/Cobas TaqMan hepatitis B virus, hepatitis C virus and human immunodeficiency virus tests. Results: Thirty-two donations were reactive by nucleic acid testing, 31 were also serologically reactive and one first-time donor was identified as having hepatitis B in the window period. Follow-up samples showed increasing titers of anti-HBs rising from 19 UI/mL in the index donation to 109 IU/mL seven months later attributable to his vaccination history. Curiously, this donor was never reactive for HbsAg nor for anti-HBc. In the yield donation, he was concomitantly reactive for syphilis (enzyme immunoassay and fluorescent treponemal antibody-absorption; venereal disease research laboratory non-reactive). Overall, six donors (0.02%) were characterized as occult hepatitis B. A total of 35% of the confirmed (recombinant immunoblot assay positive) hepatitis C donations were nucleic acid testing non-reactive and no human ...

Transplante autólogo em mieloma múltiplo: experiência de um serviço brasileiro em 15 anos de seguimento / Autologous stem-cell transplantation for multiple myeloma: a Brazilian institution experience in 15 years of follow up

Objective: To determine the 5-year post-transplant survival of patients with multiple myeloma. Methods: A retrospective study in patients diagnosed with multiple myeloma submitted to autologous bone marrow transplantation at a Brazilian institution, during the period of 1993 to 2007. Results: Seventy-three patients were evaluated with a median age of 55 years. Survival in 5 years was 75% (2.4 to 60 months). Statistical analysis demonstrated statistical significance for the applied grade of response prior to treatment with autologous bone marrow transplantation (p = 0.01). There was no statistical significance for clinical staging or time of diagnosis (before or after the year 2000). Conclusion: Experience in autologous bone marrow transplantation for multiple myeloma at a Brazilian institution demonstrated an evolution consistent with that of medical literature and highlighted the importance of a response to treatment prior to transplantation in the survival of these patients.

Objetivo: Determinar a sobrevida de pacientes portadores de mieloma múltiplo em 5 anos pós-transplante. Métodos: Estudo retrospectivo em pacientes com diagnóstico de mieloma múltiplo submetidos a transplante autólogo de medula óssea em umainstituição brasileira, no período de 1993 a 2007 . Resultados: Foram avaliados 73 pacientes com mediana de idade de 55 anos. A sobrevida encontrada em 5 anos foi de 75% (2,4 a 60 meses). A análise estatística demonstrou significância estatística para o grau de resposta ao tratamento aplicado previamente ao transplante autólogo de medula óssea (p = 0,01). Não houve significância estatística para o estadiamento clínico ou período de diagnóstico (antes ou após o ano 2000). Conclusão: A experiência na realização de transplante autólogo de medula para mileoma múltiplo, em uma instituição brasileira, demonstrou uma evolução concordante com a literatura médica e destacou a importância da resposta prévia ao transplante na sobrevida desses pacientes.

Transplante alogênico de células-tronco hematopoéticas em crianças com imunodeficiências primárias: a experiência do Hospital Israelita Albert Einstein / Allogeneic hematopoietic stem cell transplantation in children with primary immunodeficiencies: Hospital Israelita Albert Einstein experience

Objective: To report the experience of a tertiary care hospital with allogeneic hematopoietic stem cell transplantation in children with primary immunodeficiencies. Methods: Seven pediatric patients with primary immunodeficiencies (severe combined immunodeficiency: n = 2; combined immunodeficiency: n = 1; chronic granulomatous disease: n = 1; hyper-IgM syndrome: n = 2; and IPEX syndrome: n = 1) who underwent eight hematopoietic stem cell transplants in a single center, from 2007 to 2010, were studied. Results: Two patients received transplants from HLA-identical siblings; the other six transplants were done with unrelated donors (bone marrow: n = 1; cord blood:n = 5). All patients had pre-existing infections before hematopoietic stem cell transplants. One patient received only anti-thymocyte globulin prior to transplant, three transplants were done with reduced intensity conditioning regimens and four transplants were done after myeloablative therapy. Two patients were not evaluated for engraftmentdue to early death. Three patients engrafted, two had primary graft failure and one received a second transplant with posterior engraftment. Two patients died of regimen related toxicity (hepatic sinusoidal obstruction syndrome); one patient died of progressive respiratory failure due to Parainfluenza infection present prior to transplant. Four patients are alive and well from 60 days to 14 months after transplant. Conclusion: Patients' status prior to transplant is the most important risk factor on the outcome of hematopoietic stem cell transplants in the treatment of these diseases. Early diagnosis and the possibility of a faster referral of these patients for treatment in reference centers may substantially improve their survival and quality of life.

Objetivo: Relatar a experiência de um hospital terciário no tratamento de pacientes pediátricos com imunodeficiências primárias com transplante de células-tronco hematopoéticas. Métodos: De 2007 a 2010, foram realizados oito transplantes em sete pacientes pediátricos com imunodeficiências primárias: imunodeficiência combinada grave (n = 2); imunodeficiência combinada (n = 1); doença granulomatosa crônica (n = 1); síndrome hiper-IgM (n = 2); síndrome IPEX (n=1). Resultados: Dois pacientes foram transplantados com medula óssea de irmãos HLA-idênticos; seis transplantes foram feitos com doadores não aparentados (medula óssea: n = 1; sangue de cordão umbilical: n = 5). Todos os pacientes haviam tido episódios de infecção grave previamente ao tratamento. Um paciente recebeu apenas globulina antitimocítica antes do transplante de células-tronco hematopoéticas, três transplantes foram feitos com quimioterapia de intensidade reduzida e quatro após quimioterapia mieloablativa. Dois pacientes morreram precocemente e não foram avaliados em relação à enxertia. Três pacientes tiveram enxertia completa, dois evoluíram com falha primária de pega, um deles recebeu um segundo transplante com pega do enxerto. Dois pacientes morreram de toxicidade do transplante (síndrome da obstrução sinusoidal hepática), um paciente morreu de insuficiência respiratória por infecção por parainfluenza já existente antes do transplante. Quatro pacientes estão vivos e bem entre 60 dias e 14 meses após o transplante. Conclusão: A condição do paciente ao transplante é o fator mais importante no sucesso do tratamento. O diagnóstico precoce dos pacientes e a possibilidade de encaminhá-los mais rapidamente para tratamento em centros de referência podem melhorar substancialmente a sobrevida e a qualidade de vida deles.

Incidência de aloimunização eritrocitária em pacientes poli transfundidos / Prevalence of erythrocyte alloimmunization in polytransfused patients

Objective: To determine the incidence and the rate of red blood cell alloimmunization in polytransfused patients. Methods: A polytransfused patient was defined as having received at least 6 units of red cell concentrates during a 3-month period. The recordsof all patients (n = 12,904) who had received red blood cell units were examined retrospectively by searching the computer database at Hospital Israelita Albert Einstein in São Paulo, Brazil, over a 6-year period, between 2003 and 2009. Results: During this time, 77,049 red cell concentrate transfusions were performed in 12,904 patients. There were 3,044 polytransfused patients, 227 of whom (7.5%) presented with irregular erythrocyte antibodies. The prevalence of alloantibody specificity was: Anti-E>anti-D>anti-K>anti-C>anti-Dia>anti-c>anti-Jka>anti-S in 227 polytransfused patients. We found combinations of alloantibodies in 79 patients (34.8%), and the most commonspecificities were against the Rh and/or Kell systems. These antibodies show clinical significance, as they can cause delayed hemolytic transfusion reactions and perinatal hemolytic disease. About 20% of the patients showed an IgG autoantibody isolatedor combined with alloantibodies. Interestingly, a high incidence of antibodies against low frequency antigens was detected in this study, mainly anti-Dia. Conclusion: Polytransfused patients have a high probability of developing alloantibodies whetheralone or combined with autoantibodies and antibodies against low frequency antigens. Transfusion of red blood cells with a phenotype-compatible with RH (C, E, c), K, Fya, and Jka antigens is recommended for polytransfused patients in order to preventalloimmunization and hemolytic transfusion reactions.

Objetivo: Determinar a incidência e a taxa de aloimunização eritrocitária em pacientes politransfundidos. Métodos: Foram classificados como politransfundidos todos os pacientes que receberam no mínimo 6 unidades de concentrado de hemácias no período de 3 meses. Foram examinados retrospectivamente os prontuários de todos os pacientes(n = 12.904) que receberam transfusões de unidades de hemácias procurados nas bases de dados computadorizados do Hospital Israelita Albert Einstein, em São Paulo (SP), no período de 6 anos, entre 2003 e 2009. Resultados: Nesse período foram realizadas 77.049 transfusões de concentrado de hemácias em 12.904 pacientes. Os pacientes politransfundidos totalizaram 3.044, sendo que 227 (7,5%) apresentam anticorpos eritrocitários irregulares. A prevalência da especificidade dos aloanticorpos encontrados nos 227 pacientes politransfundidos foi: Anti-E>anti-D>anti-K>anti-C>anti-Dia>antic>anti-Jka>anti-S. Em 79 pacientes (34,8%) foram encontradas associações de aloanticorpos e as combinações mais freqüentes foram dos anticorpos dos sistemas Rh e/ou Kell. Esses anticorpos têm importância clínica, pois podem causar reações transfusionais hemolíticas tardias e doença hemolítica perinatal. Cerca de 20% dos pacientes apresentavam autoanticorpo IgG isolado ou em associação com aloanticorpos. Um achado interessante neste estudo foi a alta incidência de anticorpos contra antígenos de baixa frequência, com predomínio anti-Dia. Conclusão: Pacientes politransfundidos têm alta probabilidade de desenvolver aloanticorpos isolados ou em associação com autoanticorpos e anticorpos contra antígenos de baixa frequência. A transfusão de concentrado de hemácias com fenótipo compatível para os antígenos RH (C, E, c), K, Fya, e Jka deve ser recomendada para o grupo de pacientes politransfundidos, com objetivo de evitar a aloimunização e a reação transfusional hemolítica.

Fatores que afetam a coleta de células-tronco hematopoiéticas do sangue periférico por leucaférese de grande volume: experiência de um único centro / Factors affecting autologous peripheral blood hematopoietic stem cell collections by large-volume leukapheresis: a single center experience

Objective: To evaluate factors affecting peripheral blood hematopoietic stem cell yield in patients undergoing large-volume leukapheresis for autologous peripheral blood stem cell collection. Methods: Data from 304 consecutive autologous peripheral blood stem cell donors mobilized with hematopoietic growth factor (usually G-CSF), associated or not with chemotherapy, at Hospital Israelita Albert Einstein between February 1999 and June 2010 were retrospectively analyzed. The objective was to obtain at least 2 x 106 CD34+ cells/kg of body weight. Pre-mobilization factors analyzed included patient?s age, gender and diagnosis. Post mobilization parameters evaluated were pre-apheresis peripheral white blood cell count, immature circulating cell count, mononuclear cell count, peripheral blood CD34+ cell count, platelet count, and hemoglobin level. The effect of pre and post-mobilization factors on hematopoietic stem cell collection yield was investigated using logistic regression analysis (univariate and multivariate approaches). Results: Premobilization factors correlating to poor CD34+ cell yield in univariate analysis were acute myeloid leukemia (p = 0.017) and other hematological diseases (p = 0.023). Significant post-mobilization factors included peripheral blood immature circulating cells (p = 0.001), granulocytes (p = 0.002), hemoglobin level (p = 0.016), and CD34+ cell concentration (p < 0.001) in the first harvesting day. However, according to multivariate analysis, peripheral blood CD34+ cell content (p < 0.001) was the only independentfactor that significantly correlated to poor hematopoietic stem cell yield. Conclusion: In this study, peripheral blood CD34+ cell concentration was the only factor significantly correlated to yield in patients submitted to for autologous collection.

Objetivo: Avaliar fatores que afetam o rendimento da coleta em pacientes submetidos à leucaférese de grande volume para obtenção de células-tronco hematopoiéticas do sangue periférico para autotransplante. Métodos: Análise retrospectiva de 304 doadores de células-tronco hematopoiéticas de sangue periférico para autotransplante submetidos à mobilização com fator de crescimento hematopoiético (geralmente G-CSF), associado ou não à quimioterapia, no Hospital Israelita Albert Einstein de Fevereiro de 1999 a Junho de 2010. O objetivo da coleta foi obter pelo menos 2x106 CD34+ células/kg peso. Os fatores pré-mobilização incluíam idade, sexo e diagnóstico do paciente. Os parâmetros pós-mobilização avaliados foram contagem de leucócitos, células imaturas, células mononucleares e células CD34+, plaquetas e nível de hemoglobina no sangue periférico. O efeito desses fatores no rendimento da coleta de CTH foi investigado por meio de regressão logística (análise univariada e multivariada). Resultados: A análise univariada revelou os seguintes fatores pré-mobilização estatisticamente significantes: diagnóstico de leucemia mieloide aguda (p = 0,017) e outras doenças hematológicas (p = 0,023), células imaturas circulantes (p = 0,001), granulócitos (p = 0,002), nível de hemoglobina (p = 0,016) e contagem de células CD34+ (p <0,001) no primeiro dia de coleta. Entretanto, só a contagem de células CD34+ no sangue periférico manteve-se associada de forma significante ao rendimento ruim da coleta de células-tronco hematopoiéticas na análise multivariada. Conclusão: Neste estudo, a contagem de células CD34+ no sangue periférico foi o único fator significantemente associado ao rendimento da coleta de células-tronco hematopoiéticas com leucaférese de grande volume para autotransplante.

Uso de gemtuzumabe ozogamicina combinado com quimioterapia convencional em pacientes com leucemia mieloide aguda / Use of gemtuzumab ozogamycin combined with conventional chemotherapy in patients with acute myeloid leukemia

Objective: To analyze the outcome of patients treated with gemtuzumab ozogamycin combined with conventional therapy treated at Hospital Israelita Albert Einstein. Methods: 14 patients who had high risk features (secondary leukemia, unfavorable cytogenetics, and refractory disease) were treated with gemtuzumab ozogamycincombined with conventional therapy and their outcome was analysed by reviewing their medical records. Results: Overall response rate was 58%, with 43% achieving complete response, with a median followup of 11 months, event-free survival was 3 months. Eleven patients died, 6 of them due to refractory acute myeloid leukemia. Only four patients presented with grade 3 to 4 toxicities and only one patient had sinusoidal obstruction syndrome after bone marrow transplant. Conclusion: gemtuzumab ozogamycin combined with chemotherapy is a feasible treatment regimen in acute myeloid leukemia patients. However, further studies are necessary to clarify which subgroup of patients may benefit from this treatment.

Objetivo: Analisar a evolução de pacientes tratados com gemtuzumabe ozogamicina combinado à terapêutica convencional no Hospital Israelita Albert Einstein. Métodos: 14 pacientes que tinham alto risco (leucemia secundária, citogenética desfavorávele doença refratária) foram tratados com gentuzumabe ozogamicina associado à terapêutica convencional, e sua evolução foi analisada por meio de seus prontuários médicos. Resultados: A taxa total de resposta foi de 58%, com 43% chegando a resposta completa, em acompanhamento médio de 11 meses, e três meses com intervalo de sobrevivência livre. Foram a óbito 11 pacientes, 6 deles por leucemia mieloide aguda. Somente quatro pacientes apresentaram graus 3 a 4 de toxicidade e apenas um paciente teve síndrome de obstrução sinusoidal após transplante de medula. Conclusão: Gemtuzumabe ozogamicina associado à terapêutica quimioterápica convencional éum tratamento factível em pacientes com leucemia mieloide aguda. Contudo, novos estudos são necessários para esclarecer qual o subgrupo de pacientes que pode se beneficiar desse tratamento.

Transplante de células-tronco de doador não aparentado com sucesso após transplante de células de cordão umbilical não aparentado em paciente com leucemia linfocítica aguda: À procura do efeito enxerto versus leucemia: [carta ao editor] / Successful stem cell transplantation from an unrelated donor after an unrelated cord blood transplantation in an acute lymphocytic leukemia patient: searching for the graft versus leukemia effect: [letter to editor]

Segundo transplante homólogo é uma opção terapêutica parapacientes com leucemia linfoblástica aguda (LLA) que têmrecorrência após o primeiro transplante. Entretanto, para pacientessem parentes compatíveis, pode ser difícil encontrar umsegundo doador não aparentado. Uma moça de 19 anos com pro-B LLA desde 1997 recebeu quimioterapia e entrou em remissãocompleta (RC) em quatro semanas. Alta dose de citarabina foimantida por dois anos. Dois anos após o final do tratamento, apaciente apresentou recorrência, e recebeu o protocolo Hyper-CVAD (ciclofosfamida, doxorrubicina, vincristina e dexametasona).Um doador de cordão umbilical compatível 6/6 foi identificado eusado. Não havia sinais de doença do enxerto versus hospedeiro(DEVH) aguda. Segunda recorrência foi diagnosticada 32 mesesapós o transplante. O tratamento consistiu novamente no protocoloHyper-CVAD), sem resposta, e, subsequentemente, citarabina dealta dosagem com mitoxantrona de alta dosagem, resultando numaterceira RC. Um doador não aparentado, 10/10 HLA compatível,foi identificado. A despeito da terapia profilática, a pacientedesenvolveu DEVH aguda e crônica e múltiplos episódios deinfecções, mas se recuperou completamente. Ela está viva, emboa performance e em remissão completa 24 meses após o segundotransplante. Houve tempo para encontrar um doador nãoaparentado após o transplante inicial de células de cordão apenasporque a paciente entrou numa terceira RC após quimioterapia.Segundo transplante de célula-tronco alogênico, de doador nãoaparentado, pode ser oferecido a pacientes selecionados apóstransplante de células de cordão, especialmente após recorrência,sem DEVH e/ou o fenômeno enxerto versus leucemia.

Aplicações da genotipagem de grupos sanguíneos / Applications of blood group genotyping

A possibilidade de realizar tipagem de grupos sangüíneos por DNA pode facilitar a resolução de problemasclínicos que não são solucionados pela hemaglutinação, aumentando assim a segurança dos pacientes transfundidos. Agenotipagem de grupos sangüíneos pode ser aplicada nas seguintes situações (a) tipagem de antígenos para os quais não há disponibilidade de anti-soros comerciais, (b) tipagem de pacientesrecentemente transfundidos e/ou que apresentam teste direto da antiglobulina positivo, (c) identificação de fetos de risco de mães RhD-negativo, (d) tipagem de doadores para aumentar o estoque de sangue raro para as transfusões. O objetivo desta revisão é apresentar as aplicações da genotipagem de grupos sangüíneos na medicina transfusional e materno-fetal e discutir os problemas clínicos que potencialmente podem ser resolvidospelas técnicas moleculares. A base de dados utilizada nesta revisão foi a busca no MEDLINE e por meio de revisão das referências bibliográficas. Todos os trabalhos e revisões incluídos abrangeram a segurança transfusional e materno-fetal, as bases moleculares dos antígenos de grupos sangüíneos e as aplicações da genotipagem na práticaclínica. Os dados foram coletados através da avaliação dos resultados de estudos e revisões de polimorfismosde grupos sangüíneos e suas aplicações na clínica.