INTRODUCTION:Dendritic cell (DC) vaccines have demonstrated good efficacy in preventing relapse and in increasing survival of patients affected by a variety of both solid and hematological tumors. Most protocols used to generate these cells involve the automated separation of peripheral blood monocytes from patients. This approach requires specialized equipment, which elevates the cost of this type of therapy, potentially limiting the widespread access to patients.METHOD:In this study, we compare the yield and quality of dendritic cells generated from monocytes and isolated by an automated method or by manual methods using gradient centrifugation.RESULTS:The results demonstrate the equivalence of the 3 methods in relation to the yield and final quality of the product, however with considerable differences between the costs of these procedures. In addition, this study also demonstrates the feasibility of the antigenic pulse with autologous tumor cell lysates, constituting a source of antigens, not only easily obtained and manipulated, but also specific to the patient's tumor.CONCLUSION:These findings may have important implications for emerging centers interested in using this medical approach and potentially increase the access of a greater number of patients to this therapeutic option.

Background RBC alloimmunization remains a significant problem for many patients with SCD. To reduce alloimmunization some strategies have been implemented to provide limited or extended antigen matched RBC transfusions to patients with SCD who need chronic transfusion support. The aim of this study was to evaluate the effects of prophylactic RBC transfusion with extended antigen matching on alloimmunization in patients with SCD. Methods This is a 20-year retrospective study of patients with SCD transfused with RBCS that were prospectively matched for D, C, c, E, e, K, Fya/Fyb, Jka/Jkb and S antigens. Our study included 95 patients, and none had antibodies documented before their first transfusion. Patients and donors were phenotyped and molecular typing was performed in all patients who had recent transfusions or a positive direct antiglobulin test to predict their antigen profile. Unexpected antibodies to the Rh system, meaning anti-Rh antibodies in patients whose serologic phenotype was Rh positive, were investigated by molecular genotyping for RH variant alleles. Results During this study-period, 12 (12.6%) were alloimmunized and 83 (87.4%) were not. Among the 12 patients who alloimmunized, 7 (58.3%) developed antibodies to Rh antigens and 5 (41.7%) produced antibodies to low prevalence antigens. All patients who developed Rh antibodies had RH variant alleles. Autoantibodies were found in 16 (16.8%) transfused patients. Conclusion SCD patients benefit from receiving prophylactic RBC transfusions with extended antigen matching, as demonstrated by the reduction on the rates of alloimmunization and the lack of antibodies to K, FY, JK and S antigens, however, this strategy does not avoid alloimmunization to Rh and low-prevalence antigens.

Limb ulcers are one of the most debilitating complications of Sickle Cell Diseases (SCD), greatly limiting the quality of life of these patients. Although Hydroxyurea (HU) is a fundamental element in the management of patients with SCD, its role of in the emergence or worsening of limb ulcers in SCD patients is still a matter of debate.

Disturbances in the physiological regulation of erythropoietin (EPO) in patients with sickle cell disease (SCD) may contribute to worsening anaemia and increased transfusion requirements, but the use of recombinant EPO in this group of patients is controversial. The objective of this study was to evaluate the use of this drug in adult patients with SCD and its effects on haemoglobin levels and transfusion requirements. We conducted a retrospective analysis at the University of Campinas, with nineteen adults with a diagnosis of SCD (HbSS and HbS/β+ thalassaemia), who had received at least 1 year of EPO therapy between 2007 and 2014. Haemoglobin concentrations and trends of variation in transfused RBC volumes were compared before and after EPO administration. We observed that seven patients had a good response to treatment (Hb increment higher than 1·5 g/dl) and nine had a partial response (0·5–1·5 g/dl increment) and there was a significant decrease in the need for transfusion amongst those who usually required regular transfusions. There were no increases in the rates of vaso‐occlusive crisis or venous thromboembolism in comparison to the year before the onset of the therapy. Erythropoietin therapy led to a marked increase in haemoglobin concentration with a concomitant decrease in the demand for transfusion. Considering all complications related to allogeneic transfusion, we believe that EPO therapy represents an important therapeutic tool in sickle cell anaemia.

O envolvimento renal na doença falciforme compreende uma gama de desordens glomerulares e tubulares sendo sua incidência diretamente proporcional ao aumento da sobrevida dessa população sendo importante causador de morbidade e mortalidade desses pacientes. Os distúrbios renais nas doenças falciformes podem apresentar-se de forma assintomática, ou através de sinais como hematúria. Envolvidos na fisiopatologia desses eventos estão diversos mecanismos, como alterações na concentração e acidificação urinárias, microtromboses e infarto renal. A isquemia medular pode levar a inabilidade em manter o gradiente de íon hidrogênio, levando a uma forma incompleta de acidose tubular renal distal (tipo IV), que pode estar associada a hipercalemia independente da aldosterona. Há escassez de dados na literatura avaliando hipercalemia e acidose tubular renal especificamente em pacientes portadores de síndromes falciformes. Pouco se sabe também sobre a melhor maneira de avaliar a função renal nesse grupo de pacientes, cuja doença é caracterizada, nos estágios iniciais, por hiperfluxo glomerular, levando a falsas estimativas do real acometimento da função dos rins. Há escassez de dados na literatura avaliando hipercalemia e acidose tubular renal especificamente em pacientes portadores de síndromes falciformes. O objetivo deste estudo é fazer esse diagnóstico, e analisar a eficiência do N-GAL(Neutrophil Gelatinase Associated Lipocalin) para este diagnóstico.

OBJECTIVES:Deferasirox is an oral iron chelator with established dose-dependent efficacy for the treatment of iron overload secondary to transfusion. However, there is few data reporting the use of Desferasirox in adult patients with sickle cell disease (SCD) and transfusional iron overload. METHODS:We conducted a prospective, single center, nonrandomized study from January 2014 to March 2015 in Campinas, Brazil. Seven patients (five women, median age 50 y.o.) who were followed up on regular transfusion program were treated with a single daily dose of deferasirox (median dose 20 mg/kg). They were monitored for clinical symptoms, renal function and hepatotoxicity. RESULTS:One patient discontinued the study due to lack of compliance. Two patients reported mild to moderate adverse events (gastrointestinal disturbances). Five patients had the drug discontinued due to worsening of renal function. One patient had the drug discontinued due to severe hepatotoxicity that evolved to death; no patient finished the study. Discussion and conclusions: Deferasirox does not appear to be well tolerated in SCD patients older than 40 years, in which complications of the underlying disease are already fully installed. The choice of the ideal iron chelator for this population should include an evaluation of comorbidities and organic dysfunctions, as well as the need to find pharmacogenetic safety markers in this group of patients.

Platelet transfusion is an essential supportive therapy for patients with hemato-oncological disorders as many of them present with thrombocytopenia and bleeding.Platelet refractoriness, defined as the lack of adequate post-transfusion platelet count increment, remains a challenge in the management of platelet transfusion dependent patients.The majority of refractory cases have nonimmune causes and include infection/sepsis, fever, splenomegaly, use of antibiotics and even storage conditions. 1 Immune causes comprise ABO incompatibility, antibodies against class I human leukocyte antigens (HLA) (mostly HLA-A and HLAB), and antibodies against human platelet antigens (HPA).In hematology/oncology patients, platelet refractoriness has been reported in 7-34% of cases. 2 The transfusion needs of patients with immune platelet refractoriness triggers a series of processes that include the identification of the immune cause, the search for and notification of compatible donors in a genotyped donor bank, the donation of the actual blood product and finally its release and availability for use.These are lengthy procedures, which are costly and often unsuccessful.On the other hand, we have the refractory patient, bleeding or at imminent risk of bleeding, where urgency and agility are essential.These are two dissonant situations attempting to converge on the same goal.In this regard, despite the few studies that rigorously assess bleeding in patients requiring cross-matched platelet support, the refractory state is associated with increased mortality. 3hus, the development of technologies that aim to optimize the availability of the blood product as described by

Objectives and methods: We evaluated possible relationships between echocardiographic findings and clinical and laboratory parameters, in a cohort of Brazilian patients diagnosed with sickle cell/beta-thalassemia, to better understand the cardiac involvement in this disease. Results: Left atrial (LA) and left ventricular (LV) dilation were found in 19.5 and 11% of patients, respectively; systolic left ventricular dysfunction was present in a single patient. There were no differences in masses and volumes of cardiac chambers comparing S beta(0) with S beta(+) patients, and no relationship between these parameters and specific complications of the disease. However, parameters of altered ventricular geometry were significantly correlated with serum creatinine, hepatic transaminases and bilirubin levels. Moreover, 3 patients presented stroke; they were significantly older [53 (41-56) x 37.5 (18-70), p = 0.048], had higher values of LV posterior wall diastolic thickness [10 (10-11) x 8 (6-14), p = 0.03], LV mass [226 (194-260) x 147 (69-537), p = 0.039] and LA/aortic ratio [1.545 (1.48-1.61) x 1.26 (0.9-1.48), p = 0.032]. Conclusions: Cardiac involvement in this disease does not appear to depend on the thalassemia phenotype. The presence of signs of myocardial remodeling in this group of patients was related to multi-organ impairment and rendered a higher propensity for stroke in older patients, suggesting the need for greater vigilance and control of associated factors. (C) 2018 Associacao Brasileira de Hematologia, Hemoterapia e Terapia Celular. Published by Elsevier Editora Ltda. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

This manuscript describes the case of a patient with sickle cell anemia who died of fulminant hepatitis after therapy with the iron chelator Deferasirox. The patient was homozygous for the -1774delG polymorphism in the Abcc2 gene, which raises the concern about the use of hepatotoxic drugs in this specific context.

BACKGROUND The reason for the difference in susceptibility to red blood cell (RBC) alloimmunization among patients with sickle cell disease (SCD) is not clearly understood and is probably the result of multiple factors. Our hypothesis is that genetic polymorphisms are associated with RBC alloimmunization. STUDY DESIGN AND METHODS We investigated the possible association of susceptibility to RBC alloimmunization with polymorphisms of HLA and cytokines genes in 161 SCD patients prior exposed to RBC transfusion. Cytokine gene polymorphisms were analyzed by polymerase chain reaction (PCR) and TaqMan assays. HLA Class I genotyping was performed using PCR‐specific sequence of oligonucleotides. Polymorphism frequencies were compared using the Fisher's exact test. RESULTS Our results revealed increased percentage of the A allele and the GA genotype of the TNFA −308G/A cytokine among alloimmunized patients when compared to nonalloimmunized patients (A allele, 16.4% vs. 6.8%, p = 0.004; GA genotype, 32.8% vs. 11.7%, p = 0.0021). In addition, the IL1B −511T allele and the IL1B −511TT and CT genotype frequencies were overrepresented among alloimmunized patients (T allele, 53.0% vs. 37.5%, p = 0.0085; CT + TT genotypes, 81.82% vs. 60.87%, p = 0.0071). In relation to HLA Class I, we found a higher frequency of HLA‐DRB1*15 among patients alloimmunized to Rh antigens when compared to nonalloimmunized patients (15.63% vs. 6.98%, p = 0.044). CONCLUSION Brazilian SCD patients with the TNFA , IL1B, and HLA‐DRB1 gene polymorphisms were at increased risk of becoming alloimmunized by RBC transfusions. These findings may contribute to the development of future therapeutic strategies for patients with SCD with higher susceptibility of alloimmunization.

O envolvimento renal na doença falciforme compreende uma gama de desordens glomerulares e tubulares sendo sua incidência diretamente proporcional ao aumento da sobrevida dessa população sendo importante causador de morbidade e mortalidade desses pacientes.Os distúrbios renais nas doenças falciformes podem apresentar-se de forma assintomática, ou através de sinais como hematúria.Envolvidos na fisiopatologia desses eventos estão diversos mecanismos, como alterações na concentração e acidificação urinárias, microtromboses e infarto renal.A isquemia medular pode levar a inabilidade em manter o gradiente de íon hidrogênio, levando a uma forma incompleta de acidose tubular renal distal (tipo IV), que pode estar associada a hipercalemia independente da aldosterona.Há escassez de dados na literatura avaliando hipercalemia e acidose tubular renal especificamente em pacientes portadores de síndromes falciformes.Pouco se sabe também sobre a melhor maneira de avaliar a função renal nesse grupo de pacientes, cuja doença é caracterizada, nos estágios iniciais, por hiperfluxo glomerular, levando a falsas estimativas do real acometimento da função dos rins.O objetivo deste estudo é avaliar a prevalência da acidose tubular renal em coorte específica de pacientes brasileiros com anemia falciforme atendidos no Hemocentro de Campinas e identificar exames preditivos dessa alteração.

There are 99 hemoglobin (Hb) variants with increased oxygen affinity inducing compensatory erythrocytosis (Globin Gene Server Web Site: http://globin.cse.psu.edu). One of these mutations, named Hb Coimbra, was described in 1991 in Portugal by Tamagnini et al. (HBB c.300T>A, p.Asp100Glu). Limited amount of published clinical and laboratory data of this condition and the absence of specific guidelines for its management suggest it would be a very benign condition. Here we report clinical and laboratorial findings of carriers of Hb Coimbra with some unexpectedly more severe clinical presentations.

BACKGROUNDPregnancy represents a challenge for women with sickle cell disease (SCD), with higher rates of both maternal and fetal complications. The aim of this study was to evaluate the impact of prophylactic transfusion support administered specifically to pregnant women with sickle hemoglobin C disease.MATERIALS AND METHODSPatients were divided into two groups according to the type of transfusion support received: 10 women received prophylactic erythrocytapheresis or manual exchange transfusion at 28 weeks of gestation, and 14 received transfusions only on demand, due to acute complications, or received no transfusions at all.RESULTSOur results indicated higher frequencies of SCD‐related complications in the group that did not receive prophylactic transfusion support (35.7% vs. only 10% in the erythrocytapheresis group). Furthermore, these complications were more severe in this group and included all cases of acute chest syndrome. A significant difference was observed concerning gestational age at birth (38.7 weeks in the transfusion group vs. 34.4 weeks, p = 0.037), with a higher frequency of preterm births in the nontransfused group (69.23% vs. 30% in the transfusion group).CONCLUSIONWe demonstrated a clear reduction of unfavorable outcomes in patients receiving prophylactic transfusions, probably reflecting better maternal and fetal conditions, which corroborated to the more satisfactory indices of vitality, observed in newborns. Considering that erythrocytapheresis or manual exchange transfusions both represent feasible and safe procedures, they could represent important tools for the optimal management of these patients.

OBJECTIVES:In sickle cell/β-thalassemia, mutations in the corresponding β-globin genes are responsible for complex pathological events resulting in diverse clinical complications. The objective of this study was to provide an overview of the clinical and laboratory characteristics of patients with the syndrome, and of the degree of severity of clinical manifestations resulting from the β-thalassemia mutation.METHODS:A retrospective chart review was performed on 46 patients with sickle cell/β-thalassemia (31 Sβ° and 15 Sβ+), evaluating hematological parameters and end organ damage. Statistical analyzes were carried out in order to highlight differences between the two groups according to the nature of the thalassemia mutation.RESULTS:As expected, patients with the Sβ0 phenotype had a higher degree of hematological involvement in comparison to Sβ+ patients; with lower hemoglobin levels, and signs of more intense chronic hemolysis. However, Sβ+ patients were more prone to the occurrence of acute chest syndrome. The impact of the thalassemia mutation upon total body and bone composition was also evident, as Sβ0 patients presented lower body mass index (BMI) and bone mineral density. The degree of bone damage correlated to lower BMI and hemoglobin levels, as well as plaquetosis, monocytosis and elevated lactate dehydrogenase, possibly reflecting the effects of hemolysis and inflammation upon bone metabolism and body constitution.CONCLUSIONS:This study identified significant differences among sickle cell/β-thalassemia patients according to the beta mutation involvement, pointing to an important predictor of disease severity.

Purpose: Despite not yet explored, the serum lactate dehydrogenase (LDH) level in hemoglobinopathy SC (HbSC) patients could be a marker of disease severity as this association is strong in sickle cell patients. We hypothesized that the degree of hemolysis in HbSC patients is a key determinant influencing a spectrum of complications that reflect the severity of HbSC vasculopathy. The aim of this study was to analyze the associations between hemolytic parameters and chronic complications in adult SC patients.Findings: We demonstrated that LDH reflects the overall rate of hemolysis and presents a correlation with the complications related to the hemolytic subphenotype: retinopathy, venous thromboembolism and leg ulcers in HbSC patients. Remarkably, this simple biomarker was associated with a clinical subphenotype of complications in patients with HbSC disease.Conclusions: We propose that LDH elevation identifies HbSC patients with hemolysis, which could be a marker of endothelial dysfunction and end-organ vasculopathy. The use of this test as a marker of disease severity could complement the decisions taken during HbSC patient management. (C) 2016 Elsevier Inc. All rights reserved.

Objectives: In sickle cell/beta-thalassemia, mutations in the corresponding beta-globin genes are responsible for complex pathological events resulting in diverse clinical complications. The objective of this study was to provide an overview of the clinical and laboratory characteristics of patients with the syndrome, and of the degree of severity of clinical manifestations resulting from the beta-thalassemia mutation.Methods: A retrospective chart review was performed on 46 patients with sickle cell/beta-thalassemia (31 S beta degrees and 15 S beta(+)), evaluating hematological parameters and end organ damage. Statistical analyzes were carried out in order to highlight differences between the two groups according to the nature of the thalassemia mutation.Results: As expected, patients with the S beta(0) phenotype had a higher degree of hematological involvement in comparison to S beta(+) patients; with lower hemoglobin levels, and signs of more intense chronic hemolysis. However, S beta(+) patients were more prone to the occurrence of acute chest syndrome. The impact of the thalassemia mutation upon total body and bone composition was also evident, as S beta(0) patients presented lower body mass index (BMI) and bone mineral density. The degree of bone damage correlated to lower BMI and hemoglobin levels, as well as plaquetosis, monocytosis and elevated lactate dehydrogenase, possibly reflecting the effects of hemolysis and inflammation upon bone metabolism and body constitution.Conclusions: This study identified significant differences among sickle cell/beta-thalassemia patients according to the beta mutation involvement, pointing to an important predictor of disease severity.