Cytomegalovirus (CMV) genotype in allogeneic hematopoietic stem cell transplantation.

BACKGROUND: Based on sequence variation in the UL55 gene that encodes glycoprotein B (gB), human cytomegalovirus (CMV) can be classified into four gB genotypes. Previous studies have suggested an association between CMV gB genotype and clinical outcome in patients who underwent an allogeneic hematopoietic stem cell transplant (HSCT). The goals of this study were identify patients with active infection caused by CMV in recipients of HSCT; determine the prevalence of CMV genotypes in the study group; correlate genotype with CMV disease, acute GVHD and overall survival. METHODS: The diagnosis of active CMV infection after allogeneic HSCT was detected by antigenemia (AGM) and/or nested-PCR (N-PCR). Positive samples from patients with active CMV infection were submitted to genotyping using N-PCR to amplify a region of UL55, followed by restriction analysis based on HinfI and RsaI digestion. Real-time PCR (qPCR) was used to determine the viral load during active CMV infection and antiviral treatment. RESULTS: Sixty-three allogeneic HSCT recipients were prospectively evaluated; 49/63 (78%) patients were infected with CMV genotypes - gB1 19/49 (39%), gB2 17/49 (35%), gB3 3/49 (6%), gB4 7/49 (14%) - and 3 (6%) had mixed CMV genotypes (gB1 + gB3, gB1 + gB4 and gB2 + gB4). Characterized by gastrointestinal disease, CMV disease occurred in 3/49 (6.1%) patients, who had CMV gB3 genotype. These gB3 genotype patients presented an increasing AGM number, mean 125 (± 250) (P = 0.70), and qPCR copies/ml, mean 37938 (SD ± 50542) (P = 0.03), during antiviral treatment, when compared with other CMV genotypes. According to CMV genotypes, stratified overall survival was 55% for gB1, 43% for gB2; 0% for gB3 and 57% for gB4 (P = 0.03). CONCLUSIONS: One of the restrictions of the presented study was the low number of CMV gB sub-cohorts). However, we demonstrated that the frequency of active CMV infection in this HSCT population was high, and the most prevalent genotype in these patients with active CMV infection was gB1 and gB2 genotype (74%). In Brazil, HSCT recipients seem to carry mainly gB1 and gB2 CMV genotype.

Time-course of sFlt-1 and VEGF-A release in neutropenic patients with sepsis and septic shock: a prospective study.

BACKGROUND: Septic shock is the most feared complication of chemotherapy-induced febrile neutropenia. So far, there are no robust biomarkers that can stratify patients to the risk of sepsis complications. The VEGF-A axis is involved in the control of microvascular permeability and has been involved in the pathogenesis of conditions associated with endothelial barrier disruption such as sepsis. sFlt-1 is a soluble variant of the VEGF-A receptor VEGFR-1 that acts as a decoy receptor down-regulating the effects of VEGF-A. In animal models of sepsis, sFlt-1 was capable to block the barrier-breaking negative effects of VEGF-A and to significantly decrease mortality. In non-neutropenic patients, sFlt-1 has been shown to be a promising biomarker for sepsis severity. METHODS: We prospectively evaluated concentrations of sFlt-1 and VEGF-A at different time-points during febrile neutropenia, and evaluated the association of these levels with sepsis severity and septic shock development. RESULTS: Neutropenic patients that evolved with septic shock (n = 10) presented higher levels of sFlt-1 and VEGF-A measured 48 hours after fever onset than patients with non-complicated sepsis (n = 31) and levels of these biomarkers correlated with sepsis severity scores. Estimation of the diagnostic accuracy of sFlt-1 levels for the discrimination of patients that evolved to septic shock yielded promising results in our study population. DISCUSSION: Our data suggest that sFlt-1 and VEGF-A could be useful biomarkers for sepsis severity in patients with febrile neutropenia. In addition, the kinetics of sFlt-1 release in patients that evolve to septic shock suggest that the sFlt-1 could be a salvage compensatory mechanism in patients with septic shock, but that the magnitude of the sFlt-1 release observed in human sepsis is not sufficient to reproduce the beneficial anti-VEGF-A effects observed in animal models of sepsis.

Surveillance of active human cytomegalovirus infection in hematopoietic stem cell transplantation (HLA sibling identical donor): search for optimal cutoff value by real-time PCR.

BACKGROUND: Human cytomegalovirus (CMV) infection still causes significant morbidity and mortality after allogeneic hematopoietic stem cell transplantation (HSCT). Therefore, it is extremely important to diagnosis and monitor active CMV infection in HSCT patients, defining the CMV DNA levels of virus replication that warrant intervention with antiviral agents in order to accurately prevent CMV disease and further related complications. METHODS: During the first 150 days after allogeneic HSTC, thirty patients were monitored weekly for active CMV infection by pp65 antigenemia, nested-PCR and real-time PCR assays. Receiver operating characteristic (ROC) plot analysis was performed to determine a threshold value of the CMV DNA load by real-time PCR. RESULTS: Using ROC curves, the optimal cutoff value by real-time PCR was 418.4 copies/104 PBL (sensitivity, 71.4%; specificity, 89.7%). Twenty seven (90%) of the 30 analyzed patients had active CMV infection and two (6.7%) developed CMV disease. Eleven (40.7%) of these 27 patients had acute GVHD, 18 (66.7%) had opportunistic infection, 5 (18.5%) had chronic rejection and 11 (40.7%) died - one died of CMV disease associated with GVHD and bacterial infection. CONCLUSIONS: The low incidence of CMV disease in HSCT recipients in our study attests to the efficacy of CMV surveillance based on clinical routine assay. The quantification of CMV DNA load using real-time PCR appears to be applicable to the clinical practice and an optimal cutoff value for guiding timely preemptive therapy should be clinically validated in future studies.

Brazilian experience using high-dose sequential chemotherapy followed by autologous hematopoietic stem cell transplantation for relapsed or refractory Hodgkin lymphoma.

PURPOSE: We evaluate the effectiveness and toxicity of high-dose sequential chemotherapy (HDS) as salvage therapy in patients with advanced-stage Hodgkin lymphoma. PATIENTS AND METHODS: We performed a retrospective analysis on 77 patients receiving HDS between 1998 and 2006. Patients enrolled were in disease progression or relapsed disease, or did not achieve a complete remission after first-line treatment. HDS consisted of the sequential administration of cyclophosphamide and granulocyte colony-stimulating factor with stem cell harvesting, followed by methotrexate plus vincristine and etoposide. RESULTS: The majority of patients had stage III/IV (64%) and B symptoms (71.4%). Disease status improvement after HDS was observed in 24 of 57 patients (42%) previously in disease progression or relapse. HDS-related deaths occurred in 8 of 77 patients (10.4%). Four patients (5.2%) developed acute myeloid leukemia/myelodysplastic syndrome. Overall, disease-free and progression-free survival was 27%, 57%, and 25%, respectively. CONCLUSION: Despite the treatment-related mortality, HDS is feasible, with satisfactory response rates, even in patients with poor prognosis.

Brazilian experience using high-dose sequential therapy (HDS) followed by autologous hematopoietic stem cell transplantation (ASCT) for malignant lymphomas / Experiência brasileira utilizando terapia sequencial de alta dose seguido de transplante autólogo de célula-tronco hematopoética para linfomas malignos

Using the overall survival (OS), disease free survival (DFS) and progression free survival (PFS), as well as associated toxicity, the purpose of this work was to evaluate the effectiveness of HDS followed by ASCT as salvage therapy. A retrospective analysis was performed of 106 patients with high grade non-Hodgkin lymphoma receiving HDS followed by ASCT, between 1998 and 2006. Median age was 45 years (Range: 8-65), with 66 (62 percent) men. Histopathological classification was: 78 percent DLBCL patients, 12 percent T and anaplastic and 9 percent Mantle cell lymphomas; 87 percent had B cell and 12 percent T cell lymphomas; 83 percent were stage III-IV (Ann Arbor Staging), 63 percent had B symptoms, 32 percent had bone marrow involvement, 62 percent bulky disease and 42 percent high-intermediate or high risk IPI. After HDCY, 9 patients died, 7 from toxicity and 2 from sepsis. Eighty patients underwent ASCT, 47 percent were in complete remission (CR) and 15 percent died, all from toxicity. Their OS was 45 percent over 8 years. During the follow-up, another 35 patients died [4 CR, 1 partial response (PR), 2 relapsed disease (RD) and 28 disease progression (DP)], 11 (31 percent) had not performed ASCT. OS was 37 percent; DFS was 49 percent and PFS 28 percent. OS by diagnosis was 42 percent for DLBCL, 40 percent for T-cell (8 y) and 20 percent for Mantle Cell (6 y) (P=NS). OS by B symptom patients was 22 percent vs. 58 percent (P=0.002) and PFS was 23 percent vs. 37 percent (P=0.03). Patients who achieved CR after HDCY (38) had significantly better OS and PFS (38 percent and 17 percent) than patients who remained in DP (P<0.0001). Cox Regression demonstrated therapeutic lines before HDCY (Relative risk - RR = 1.41; CI 95 percent: 1.04-1.90; P= 0.02) and PD both before (RR = 2.70; CI 95 percent: 1.49-4.91, P<0.001) and after HDCY (RR = 5.38; 95 percent CI: 2.93-9.87; P<0.0001). Conclusions: Our study suggests HDS is an efficient treatment to ...

A proposta deste trabalho foi avaliar a eficácia da HDS seguida do transplante autólogo como terapia de salvamento através da sobrevida global, livre de doença e livre de progressão bem como sua toxicidade. Realizou-se estudo retrospectivo com 106 pacientes com LNH de alto grau de malignidade entre 1998 e 2006. A mediana de idade foi 45 anos (8-65); 62 por cento homens; DLBCL, 78 por cento; 12 por cento, T e anaplásico e 9 por cento, linfoma da zona do manto; 87 por cento, células B; 83 por cento estádios III-IV; 63 por cento com sintomas B; 32 por cento com infiltração da medula óssea ao diagnóstico; 62 por cento com grande massa e 42 por cento com IPI de alto risco ou intermediário. Após alta dose de ciclofosfamida (HDCY), nove pacientes faleceram. Oitenta pacientes realizaram o transplante, sendo que 47 por cento estavam em RC e 15 por cento faleceram devido à toxicidade. A sobrevida global foi de 45 por cento em oito anos para estes pacientes. Trinta e cinco pacientes não realizaram o transplante por causas diversas. Sobrevida global para todos os pacientes foi de 42 por cento, DLBCL, 40 por cento; T-cell, 40 por cento e zona do manto, 20 por cento (P=NS). Pacientes que obtiveram RC após HDCY tiveram melhor sobrevida global e livre de progressão (38 por cento e 17 por cento, respectivamente) do que os que permaneceram em PD (P<0.0001). O modelo de Cox resultou que o número de linhas terapêuticas antes da HDCY (RR 1.41 IC 95 por cento: 1.04-1.90, P=0.02) e PD antes da HDCY (RR 2.70, IC 95 por cento: 1.49-4.91, P<0.001) e após HDCY (RR 5.38, IC 95 por cento: 2.93-9.87, P<0.0001). Nosso estudo sugere que HDS é um método eficiente de tratamento para melhorar o status e reduzir a massa tumoral. Em relação à toxicidade, é factível, especialmente em pacientes de prognóstico ruim

Correlation of IL-6 and IL-10 production following bone marrow transplantation with donor cytokine gene polymorphisms / Correlação da produção de IL-6 e IL-10 seguindo o transplante de medula óssea com os polimorfismos de genes de citocinas do doador

Several candidate gene studies have demonstrated that genetic polymorphisms in cytokine genes contribute to variations in the levels of cytokines produced and this variation may influence the occurrence and severity of complications after stem cell transplantation (HSCT). In this work we compared the serum concentrations of TNF-α, IFN-γ, IL-6, IL-10, and TGF-β1 in 13 recipients following HSCT with the TNF-308, IFNG+874, IL6-174, IL10-1082,-819,-592, and TGFB1+869,+915 polymorphisms. Serum cytokine levels were assessed using commercial ELISA kits for TNF-α, IFN-γ, IL-6, IL-10, and TGF-β1 (BioSource®, Nivelles, Belgium, Europe). Donor/recipient genotypes for these cytokine polymorphisms were analyzed by polymerase chain reaction-sequence-specific primer (PCR-SSP) with the Cytokine Genotyping Primers Kit (One Lambda , Canoga Park, CA, USA). We found correlation between the levels of IL-6 and IL-10 concentrations following HSCT and the IL6-174 and IL10-1082,-819,-592 polymorphisms, but not for other cytokines investigated in this study. Those with genotypes associated with low production of IL-6 and IL-10 produced lower levels of these cytokines than those with genotypes associated with high or intermediate production of these cytokines (P < 0.05).

Estudos de vários genes candidatos têm demonstrado que polimorfismos genéticos em genes de citocinas contribuem com variações nos níveis de citocinas produzidas e esta variação pode influenciar a ocorrência e gravidade de complicações após o transplante de células-tronco hematopoéticas (TCTH). Neste trabalho comparamos as concentrações séricas de TNF-α, IFN-γ, IL-6, IL-10 e TGF-β 1 em 13 receptores seguindo o TCTH com os polimorfismos TNF-308, IFNG+874, IL6-174, IL10-1082,-819,-592 e TGFB1+869,+915. Os níveis séricos de citocinas foram medidos usando-se kits comerciais de ELISA para TNF-α, IFN-γ, IL-6, IL-10 e TGF-β 1 (BioSource®, Nivelles, Belgium, Europe). Os genótipos de doadores/receptores para estes polimorfismos de citocinas foram analisados pela reação em cadeia da polimerase com sequências específicas de primer (PCR-SSP) com o kit Cytokine Genotyping Primers (One Lambda, Canoga Park, CA, USA). Encontramos correlação entre os níveis de IL-6 e IL-10 seguindo o TCTH e os polimorfismos IL6-174 e IL10-1082,-819,-592, mas não para outras citocinas investigadas neste estudo. Aqueles com genótipos relativos à baixa produção de IL-6 e IL-10 produziram mais baixos níveis destas citocinas que aqueles com genótipos relativos à produção alta e/ou intermediária destas citocinas (P < 0,05).

Leucemia Mielóide Crônica: transplante de medula óssea / Cronic Myeloid Leukemia: bone marrow transplantation

A única terapia curativa para Leucemia Mielóide Crônica ainda é o transplante de células hematopoéticas progenitoras (TCHP); os atuais resultados com uso do imatinibe são suficientes para indicarmos o TCHP como um tratamento de segunda ou terceira linha. A decisão de realizar TCHP existe em uma variedade de momentos: falha em se conseguir remissão hemtológica, citogenética ou molecular, ou quando se perde a melhor resposta conseguida ou por progressão da doença para uma fase avançada. Há decisão também de como transplantar. Nesta revisão apontamos algumas destas decisões e as atuais controvérsias.

Although the only curative therapy for chronic myeloid leukemia remains allogeneic stem cell transplantation (allo-SCT), the results of imatinib in newly diagnosed patients are sufficiently impressive to have displaced allo-SCT to second or third-line treatment. Patients now arrive at a decision for transplantation in a variety of disease situations: failing to achieve certain hematological, cytogenetic and molecular remission by some pre-determined timepoint, having lost a previous best response or due to progression to an advanced phase. The decision is also how to transplant. In this review article, the evidence supporting some of these decisions and current controversies are discussed.

Associação dos níveis de citocinas no pós-transplante de células-tronco hematopoiéticas com a Doença do Enxerto Contra o Hospedeiro aguda / Association of cytokine levels with acute graft versus host disease following full match allogeneic stem cell transplantation

Este estudo foi realizado para investigar se os níveis séricos de sIL-2R, TNF-alfa, IFN-gama, IL-6, IL-10 e TGF-beta1 estavam associados com o desenvolvimento de DECH (Doença do Enxerto Contra o Hospedeiro) aguda. Os níveis de citocinas foram seqüencialmente mensurados por Elisa em 13 pacientes que haviam sido submetidos ao transplante alogênico de células progenitoras hematopoiéticas. Os níveis de sIL-2R e IL-10 da 1ª a 15ª semanas pós-transplante foram significativamente maiores no grupo que desenvolveu DECH aguda que naquele sem a doença. Os níveis de sIL-2R aumentaram em direta correlação com a pega do enxerto e ao tempo do DECH aguda, enquanto os níveis de IL-10 aumentaram transitoriamente pós-transplante. A média da concentração de TNF-alfa nas primeiras semanas após o transplante foi maior no grupo que desenvolveu DECH aguda. Além disso, uma queda dos níveis de TGF-beta1 após a pega esteve significativamente associada à DECH aguda. Nenhuma correlação foi encontrada entre DECH aguda e as outras citocinas investigadas. Estes resultados suportam a idéia de que um balanço entre as citocinas derivadas de linfócitos T auxiliadores do tipo 1 e 2 pode ser importante no desenvolvimento e controle da DECH aguda. Embora os níveis de sIL-2R, TNF-alfa, IL-10 e TGF-beta1 tenham sido correlacionados com a DECH aguda, os níveis de sIL-2R ao tempo da pega podem prover um melhor parâmetro para a detecção precoce de DECH aguda após o transplante alogênico.

This study was performed to investigate whether the serum levels of sIL-2R, TNF-alpha, IFN-gamma, IL-6, IL-10, and TGF-beta1 are associated with the development of acute GVHD. Serum cytokine levels were sequentially measured by sandwich Enzyme Linked-Immuno-Sorbent Assay (Elisa) in 13 patients who had received full match allogeneic stem cell transplantation. Serum sIL-2R and IL-10 levels from the 1st to the 15th week post transplantation were significantly higher in the group who developed acute GVHD than in the group without acute GVHD. Soluble IL-2R levels increased in direct correlation to engraftment and onset of acute GVHD, while IL-10 levels increased transiently following transplantation. The mean TNF-alpha concentration in the first weeks after transplantation was augmented in the group that developed acute GVHD. Furthermore, a drop in TGF-beta1 levels after the engraftment was significantly associated to acute GVHD. No correlation was found between acute GVHD and the other evaluated cytokines. These results support the idea that a balance between cytokines derived from type 1 and type 2 T-helper cells may be important in the development and control of acute GVHD. Although sIL-2R, TNF-alpha, IL-10, and TGF-beta1 levels, correlated with acute GVHD, sIL-2R levels at the engraftment may provide a better parameter for the early detection of acute GVHD after allogeneic stem cell transplantation.

Platelet glycoprotein Ibalpha polymorphisms modulate the risk for myocardial infarction.

Platelet glycoprotein Iba (GPIba) gene polymorphisms have been reported to affect the risk of developing coronary heart disease. Here, within the GPIba gene, we determine the association between the variable number of tandem repeats (VNTR), the -5C/T Kozak sequence dimorphism, and the human platelet antigen (HPA)-2 polymorphisms with occurrence of myocardial infarction (MI). Patients (n=180) presenting survivors of MI were compared to 180 controls matched by age, gender, and race. Carriers of VNTR-CD genotype had a 2-fold higher risk for MI compared to controls. The prevalence of VNTR-BC was lower among patients than among controls (P=.007). These data are in agreement with recent reports of increased plug formation by human platelets containing VNTRCD but no other VNTR genotypes. Among patients, the number of vessels severely occluded was greater among carriers of the D-allele (P=.019) or VNTR-CD (P=.026) and lower among carriers of the C-allele (P=.003) or VNTR-CC (P=.0009) compared to non-carriers of these alleles. No influence was seen with the Kozak or HPA-2 polymorphisms. Determination of VNTR of the GPIba gene may prove useful for identifying high-risk individuals for MI.

The influence of the graft monocytes in the outcome of allogeneic bone marrow transplantation.

The influence of graft monocytes on graft-versus-host disease (GVHD) has not yet been established in clinical trials. To understand this association better, we evaluated the influence of bone marrow graft monocytes aiming to analyze, primarily, the correlation with acute GVHD and chronic GVHD and, secondarily, the correlation with engraftment and survival.

Terapêutica citoprotetora em pacientes tratados com químio e/ou radioterapia anti neoplásica / Cytoprotective therapy in patients treated with chemotherapy and/or antineoplasic radiotherapy

Nos últimos anos, vários agentes citoprotetores têm sido desenvolvidos para proteger células normais dos efeitos tóxicos da quimioterapia e radioterapia. O agente citoprotetor ideal seria aquele capaz de permitir a intensificaçäo da dose dos quimioterápicos; proteger um amplo espectro de órgäos e tecidos quando do tratamento com diversos fármacos quimioterápicos; conferir proteçäo específica aos tecidos normais; preservar o efeito anti-tumoral e ter pequena e/ou controlável toxicidade e efeitos colaterais. Um citoprotetor deve ser administrado antes da quimioterapia citotóxica, ao contrário dos fatores estimuladores de colônia e do Leucovorin, que säo administrados após quimioterapia como resgate à medula óssea e estimular a sua recuperaçäo. Do ponto de vista prático existem três agentes citoprotetores: dois citoprotetores quimio-específicos (Dexrazoxane e Mesna) e um citoprotetor de amplo espectro (Amifostina). Os autores discutem as principais propriedades e utilidades destas drogas utilizadas em Onco Hematologia.

Toxoplasmose cerebral após transplante alogênico de medula óssea / Central nervous system toxoplasmosis after bone marrow transplantation

As cpmplicaçöes relacionadas ao sistema nervoso central nos pacientes submetidos a transplante autólogo ou alogênico de medula óssea apresentam uma incidência média de 2 a 8 por cento, que correlacionam com o grau de imunossupressäo, doença enxerto contra o hospedeiro e a doença de base. Por este motivo observa-se maior incidência dessas complicaçöes nos tranplantes alogênicos. Dentre as mais comumentes encontradas, destacam-se as de causas infecciosas, como encefalites secundárias, o toxoplasma, aspergilus, cândida, citomegalovírus e outros vírus. Excluindo-se as infecçöes, destacam-se as hemorragias e a encefalopatia metabólica. Apesar da incidência de 3 a 8 por cento, a mortalidade dos pacientes acometidos atinge 67 por cento dos casos. Por este motivo é que torna-se importante a avaliaçäo sorológica dos pacientes previamente ao transplante e a investigaçäo dos agentes etiológicos em pacientes imunocomprometidos, após o transplante de medula óssea.

Tratamento ambulatorial com 2-clorodeoxiadenosina para doenças linfoproliferativas / Treatment of lymphoproliferatives syndromes and indolent lymphomas by 2-clorodexiadenosina in outpatient reginer

O presente estudo trata da experiência clínica do uso do 2-clorodeoxiadenosina no tratamento da tricoleucemia, síndromes linfoproliferativas e linfomas indolentes. O modo de administraçäo usado em todos os casos exceto dois, foi o da infusäo endovenosa diária por 2 horas, durante cinco dias, com ciclos mensais, num esquema ambulatorial. Dos sete casos de tricoleucemia, tratados com um único ciclo, todos tiveram regressäo da esplenomegalia e 5/7 normalizaram o hemograma. Após um segmento de 2-14 meses, todos os pacientes permaneceram com a doença controlada. Foram tratados sete casos com leucemia linfóide crônica refratária à quimioterapia oral bem como a um esquema de poliquimioterapia endovenosa. Em quatro casos houve uma remissäo parcial, mantida por 3-13 meses.Três pacientes que foram resistentes ao tratamento morreram por infecçäo. Os dois casos de leucemia pró-linfocítica receberam dois e três ciclos respectivamente, com controle da doença por 15 e seis meses respectivamente. Todos estes achados bem como uma observaçäo preliminar com resposta em dois pacientes com linfomas indolentes confirmam os dados da literatura de que o 2-CdA é um quimioterápico com boa açäo em síndromes linfoproliferativas em linfomas indolentes. Sua administraçäo é segura no regime de administraçäo usado, o que torna seu uso mais barato e prático.

Malignant lymphoma associated with hanseniasis

There are few reports in the world medical literature concerning association between leprosy and malignant lymphoma. It seams that, although defects in immune system related to leprosy are multiple and complex, no increase in incidence of malignant lymphoma among leprosy patients can be detected. This is also true for Brazil, where leprosy is an endemic disease, which poses an important public health problem. For this reason, the authors report on three cases of malignant lymphoma: one low grade, B-cell lymphoma (immunocytoma): one high grade, T-cell lymphoma and one mixed cellularity Hodgkin's disease in patients previously diagnosed as having leprosy.