Marco Bitencourt - Academia.edu (original) (raw)
Papers by Marco Bitencourt
Biology of Blood and Marrow Transplantation
Psicologia: Ciência e Profissão, 2004
Resumo: O presente estudo, objetiva investigar a representação simbólica do câncer em profissiona... more Resumo: O presente estudo, objetiva investigar a representação simbólica do câncer em profissionais da área de saúde, pacientes, familiares e população em geral. Investigou-se, as imagens simbólicas que permeiam a atitude de cada um perante a convivência com o câncer por meio de questionário. Os resultados indicaram informações ricas de subjetividade, independentemente das diferenças demográficas. A imagética interfere na percepção, na reação e na distribuição do tempo que cada um determina para a escuta da singularidade daquele que adoece com o câncer e/ou com a contaminação simbólica. Na medida em que o fantasma da morte acompanha o paciente em seus movimentos, espera-se que este estudo possa contribuir para que se busquem formas de aprimoramento do tratamento do paciente com câncer. No entanto, para ajudar o outro, é preciso, primeiramente, redimensionar a própria formação imagética; afinal, nós, profissionais da área de saúde, também somos semeadores das imagens que se reproduzem ao longo dos séculos, apesar de toda a evolução científica. Palavras-Chave: Câncer, representação simbólica, qualidade de vida.
Cytokine, 2013
Fanconi anemia (FA) is a rare disease, autosomal recessive and X linked, which is clinically pron... more Fanconi anemia (FA) is a rare disease, autosomal recessive and X linked, which is clinically prone to development of hematological abnormalities and neoplasms, especially acute myeloid leukemia. In this work IL-10 and TGF-b levels were measured on FA patients' plasma since they are the regulatory cytokines of TNF-a and INF-c which had been described to be overexpressed in this genetic disease. Our results show increased IL-10 plasma levels in 25% of FA patients studied, but levels of TGF-b within the normal range. TNF-a and INF-c were also measured and found to be increased in 24% and 23% of FA patients, respectively. However, no inverse correlation was observed between augmented levels of IL-10 and TNF or IFN-c. Patients with elevated levels of TNF-a and INF-c presented bone marrow hypocellularity. IL-10 levels did not appear to be determinant for bone marrow cellularity. These data suggest that IL-10 is also a feature of Fanconi anemia pathophysiology.
Biology of Blood and Marrow Transplantation, 2016
We report on long-term survival in 157 patients with Fanconi anemia (FA) who survived 2 years or ... more We report on long-term survival in 157 patients with Fanconi anemia (FA) who survived 2 years or longer after their first transplant with a median follow-up of 9 years. Marrow failure (80%) was the most common indication for transplantation. There were 20 deaths beyond 2-years post-transplantation with 12 of the deaths occurring beyond 5 years after transplantation. Donor chimerism was available for 149 patients; n=112 (76%) reported >95%, n=27 (18%), 90-95%, and n=8 (5%), 20-89% donor chimerism. Two patients have <20% donor chimerism. The 10- and 15-year probabilities of survival were 90% and 79%, respectively. Results of multivariate analysis showed higher mortality risks for transplantations prior to 2003 (HR: 7.87, p=0.001), chronic GVHD (HR: 3.80, p=0.004) and squamous cell carcinoma, post-transplant (HR 38.17, p<0.0001). The predominant cause of late mortality was squamous cell carcinoma with an incidence of 8% and 14% at 10 and 15 years post-transplantation, respectively, and was more likely to occur in those with chronic GVHD. Other causes of late mortality included chronic GVHD, infection, graft failure, other cancers and hemorrhage. Although most patients are disease-free and functional long-term, our data support aggressive surveillance for long periods to identify those at risk for late mortality.
Haematologica, 2004
As some patients with Fanconi s anemia (FA) present excessive telomere shortening correlating wit... more As some patients with Fanconi s anemia (FA) present excessive telomere shortening correlating with poor outcome, we investigated whether human telomerase RNA component (hTERC) mutations also play a role in telomere shortening in 115 FA patients. Only one patient was heterozygous for the G58A polymorphism. No other mutation or deletion was found. We conclude that hTERC gene mutations do not contribute to telomere shortening in FA.
Revista Brasileira De Hematologia E Hemoterapia, 2010
Revista Brasileira de Hematologia e Hemoterapia, 2003
Page 1. 239 Pasquini R et al Rev. bras. hematol. hemoter. 2003;25(4):239-246 Relato de caso / Cas... more Page 1. 239 Pasquini R et al Rev. bras. hematol. hemoter. 2003;25(4):239-246 Relato de caso / Case report Carcinoma de células escamosas em língua pós-transplante de medula óssea por Anemia de Fanconi Squamous ...
Jornal de Pediatria, 1999
INTRODUCTION: The Imerslund-Gräsbeck syndrome is a rare hereditary autosomal recessive disease, c... more INTRODUCTION: The Imerslund-Gräsbeck syndrome is a rare hereditary autosomal recessive disease, characterized by the onset of megaloblastic anemia and asymptomatic proteinuria during the first 2 years of life. OBJECTIVE: To emphasize the importance of early detection of this disorder, due to high morbidity when not correctly treated, in addition to the necessity of screening and genetic counseling of the asymptomatic family members. METHODS: The authors report two patients, male and female, 8 and 10 years old, respectively. Their past history revealed anemia and multiple blood transfusions since their infancy. They evolved with pancytopenia during childhood and diagnosis of Severe Aplastic Anemia or Fanconi Syndrome was suspected. They were referred to the Bone Marrow Transplantation Section -HC- UFPR. RESULTS: Laboratory investigations revealed pancytopenia in peripheral blood. Bone marrow aspiration showed a marked megaloblastic erythropoiesis. Twenty-four-hour urine collection revealed proteinuria (3.0 and 5.8 g/dl respectively). Cytogenetic analysis was normal. Resolution of symptoms followed replacement therapy with parenteral vitamin B12. CONCLUSIONS: The presence of megaloblastic anemia in children should be followed by investigation of proteinuria, due to the existence of this rare disorder, that has a simple diagnosis and an effective treatment.
Revista Brasileira de Hematologia e Hemoterapia, 2008
Imatinib mesylate is currently the gold-standard therapy for patients with newly diagnosed Chroni... more Imatinib mesylate is currently the gold-standard therapy for patients with newly diagnosed Chronic Myelogenous Leukemia. From the clinical trials in 1998 to the IRIS study, which compared first line imatinib treatment with interferon and low dose ara-C, this drug has been consolidated in regards to its safety and efficacy. There are still some questions to answer. Which would be the best initial dose? Are there any patients who benefit more than others? What is the best approach to suboptimal response and resistance? The most important published clinical studies are reviewed in the current article and discussed from a Brazilian perspective.
Revista Brasileira de Hematologia e Hemoterapia, 2010
... Os pacientes em ambos os grupos eram comparáveis em relação à idade, sexo, estádio, isotipo d... more ... Os pacientes em ambos os grupos eram comparáveis em relação à idade, sexo, estádio, isotipo da proteína monoclonal e nível de β2-microglobulina. ... As características e fatores de risco dos pacientes eram semelhantes entre os quatro grupos. ...
Human Mutation, 2003
Fanconi anemia (FA) is a genetically heterogeneous autosomal recessive syndrome associated with c... more Fanconi anemia (FA) is a genetically heterogeneous autosomal recessive syndrome associated with chromosomal instability, hypersensitivity to DNA cross-linking agents, and predisposition to malignancy. The gene for FA complementation group G (FANCG) was the third FA gene to be cloned, and was found to be identical with human XRCC9, which maps to 9p13. The cDNA is predicted to encode a polypeptide of 622 amino acids, with no sequence similarities to any other known protein or motifs that could point to a molecular function for FANCG/XRCC9. We used single strand conformational polymorphism analysis (SSCP) to screen genomic DNA from a panel of 307 racially and ethnically diverse unrelated FA patients from the International Fanconi Anemia Registry (IFAR) for variants in FANCG. Twenty-seven abnormal SSCP patterns were found; 18 of these variants appear to be pathogenic mutations while nine are likely to be nonpathogenic polymorphisms. Direct sequencing of genomic DNA from seven FA-G probands with one mutant allele not detected in the SSCP study and three additional probands assigned to the FA-G complementation group by retroviral correction with FANCG resulted in the detection of nine additional pathogenic mutations and two common SNPs. Conditions for rapid screening for these mutations by DHPLC for use in a clinical laboratory setting were established. The most common FANCG mutations in the IFAR population were: IVS8-2A>G (seven Portuguese-Brazilian probands), IVS11+1G>C (seven French-Acadian probands), 1794_1803del10 (seven European probands), and IVS3+1G>C (five Korean or Japanese probands). Our data suggest that the Portuguese-Brazilian, French-Acadian, and Korean/Japanese mutations were likely to have been present in a founding member of each of these populations.
European Journal of Haematology, 2005
Fanconi anaemia (FA) is a rare genetic disease whose patients have a high predisposition to haema... more Fanconi anaemia (FA) is a rare genetic disease whose patients have a high predisposition to haematological abnormalities and cancer. Fas expression levels in peripheral blood lymphocytes samples of 73 FA patients were measured to verify if alterations in Fas expression could lead to predisposition/resistance to spontaneous or PHA induced apoptosis, as well as, to reflect some haematological features of this disease. The anti- and pro-apoptotic proteins Bcl-2 and Bax were also evaluated. FA patients samples could be divided into three different groups based on Fas expression: 20 samples had low, 32 normal and 21 increased Fas levels when compared to 41 control samples. No correlation was found between Fas and Bcl-2 expression but a good association was obtained with Bax, in the subgroup with increased Fas expression. The best correlation was seen between Bax expression and apoptosis. Out of the 15 samples with high Bax expression, 11 underwent apoptosis whereas only one out of seven samples with low levels of Bax displayed increased induced apoptosis. Most patients with normal haematological features expressed Fas within normal levels. It is difficult to establish, however, if Fas-expression is involved in the cause or is a consequence of the effects observed.
Brazilian Archives of Biology and Technology, 2005
ABSTRACT
Biology of Blood and Marrow Transplantation, 2014
Biology of Blood and Marrow Transplantation, 2007
The major complication associated with allogeneic hematopoietic stem cell transplantation (HSCT) ... more The major complication associated with allogeneic hematopoietic stem cell transplantation (HSCT) is graft-versus-host disease (GVHD), which is mediated by mature T cells in the donor transplant inoculum. Since donor T cells are also important in preventing graft failure, opportunistic infections and leukemia relapses after transplant, it would be highly advantageous to identify and separate T cells with GVHD-reactivity from those capable of mediating graft-versus-leukemia (GVL) responses. In this regard, TCR V repertoire analysis by CDR3-size spectratyping can be a powerful tool to characterize alloreactive T cell responses. As an initial test of this capability, we studied lethal GVHD in the B6 -Ͼ CXB-2 MHC-matched, minor histocompatibility antigen disparate murine model to determine the correlation between the in vitro detected alloreactive B6 CD8 ϩ T cell responses and their actual involvement in the development of GVHD upon transfer to lethally irradiated CXB-2 recipients. Previous methods to obtain the V repertoire from responding mixed lymphocyte reactions (MLR) required expansion of the alloreactive cells in secondary stimulation cultures, for a total of 20 days. To significantly shorten this incubation period, in order to eventually adapt this technique for clinical use, we used the fluorescent-based intracellular dye CFSE to label responding cells in a 4-day MLR and fluorescent cell sorting to isolate the divided alloreactive cells that could then be processed for V spectratype analysis. In a comparative study of the predictive value of the V repertoire detected in traditional MLR versus CFSE/MLR, the results indicated that the former approach correlated with 78% of the V CDR3-size skewing patterns observed in mice with B6 -Ͼ CXB-2 GVHD, and the CFSE/MLR approach corresponded to 65% of those skewed V families found in vivo. These results suggest that in vitro spectratyping analysis, and particularly the use of CFSE to predict in vivo reactivity, may be a useful technique in eventually guiding the manipulation of the donor T cell inoculum in order to improve the outcome of HSCT.
Biology of Blood and Marrow Transplantation, 2007
pts engrafted at a md of 22 (11-41) days. 6 pts lost complete chimerism during follow up with pro... more pts engrafted at a md of 22 (11-41) days. 6 pts lost complete chimerism during follow up with proven relapse at a md of 401 days in four. At a md follow up of 23 (4-79) months probability of overall (OS) vs eventfree survival (EFS) was 74% vs 50% for the whole cohort. A trend towards better survival was seen in recipients of related compared to unrelated transplants (100 vs 64 %, pϭ0.06) and in those with less advanced disease (90 vs 68 % in Dupriez 0 vs 1ϩ2, pϭ0.26). However the only significant predictor for OS by log rank test was the pretransplant CCI: 85 vs 34 % in pts with CCI 0 to 2 vs 3 to 6 (pϭ0.01). Probability of EFS at day 480 was significantly less in pts with abnormal karyoptype compared to those with no detectable anomalies (17 vs 61%, pϭ0.01). Five pts with decreasing chimerism received immunotherapy (3 DLI, 2 PBSC) with return to complete chimerism in three and postDLI aplasia in one.-Conclusions: Our results support the use of related and unrelated alloHCT as a curative treatment option in MMM. Outcome is better in pts without disease associated and disease independent comorbidity. For pts with abnormal karyotype there is a considerable risk of relapse/reversal post transplant. Especially in risk pts chimerism therefore should be monitored closely as disease recurrence may respond to immunotherapy.
Biology of Blood and Marrow Transplantation, 2007
We have demonstrated that patient derived myeloma cells fused with autologous dendritic cells (DC... more We have demonstrated that patient derived myeloma cells fused with autologous dendritic cells (DCs) potently stimulate anti-tumor immunity in vitro. We are conducting phase I clinical trials in which patients with myeloma undergo serial vaccination with DC/ myeloma fusions alone or in conjunction with stem cell transplantation. To date, 18 patients have been enrolled (11-vaccine alone, 7 vaccine following autologous stem cell transplant). To generate mature DCs, adherent mononuclear cells were isolated from a leukapharesis collection and cultured with GM-CSF, IL-4, and TNF␣ . The mean yield and viability of the DC preparations was 1.5 ϫ 10 8 cells and 88%, respectively. Patient derived myeloma cells were isolated from bone marrow aspirates and were quantified by the expression of CD38 and/or CD138. The mean yield and viability of the myeloma cell collections was 7.3 ϫ 10 7 cells and 89%, respectively. Fusion cells were generated by coculture of DCs with myeloma cells in the presence of 50% polyethylene glycol. The mean fusion efficiency was 40% as determined by the percentage of cells that co-expressed unique DC and myeloma antigens. In contrast to myeloma cells, the DC and fusion cell preparations prominently stimulated allogeneic T cell proliferation in vitro. To date, 13 patients have completed vaccination at a dose of 1-5 ϫ 10 6 fusion cells. GM-CSF (100 g) was administered subcutaneously on the day of vaccination and for 3 days thereafter. Adverse events judged to be potentially vaccine related have included vaccine injection site reactions, edema, rash, fever, chills, fatigue, muscle aches, pruritis, and diarrhea. One patient with a history of prior deep venous thrombosis (DVT) developed a DVT and pulmonary embolus of uncertain relation to the vaccine. To date, a majority of evaluable patients demonstrated evidence of vaccine induced anti-myeloma immunity as demonstrated by at least 2 fold increase in IFN␥ expression by CD4 and/or CD8 T cells in response to ex vivo exposure to autologous tumor lysate. Of patients undergoing vaccine therapy alone, 5 patients demonstrated stabilization of the myeloma paraprotein for 2-6 months following initiation of vaccination. Of 3 patients completing posttransplant vaccination, 2 patients demonstrated resolution of the persisting myeloma protein post-transplant and 1 patient demonstrated a transient increase followed by a decline in paraprotein levels post-transplant.
Biology of Blood and Marrow Transplantation, 2007
sults There was no significant difference between the 3 groups in the occurrence of FN or documen... more sults There was no significant difference between the 3 groups in the occurrence of FN or documented infection. However, hyperglycemia was significantly associated with organ dysfunction and aGVHD. OS was better and TRM was less in group1 compared with group2 and group3. Conclusion Degrees of hyperglycemia during neutropenia was associated with an increased risk of organ dysfunction and aGVHD, which further led to higher TRM and lower OS. These results support the possibility that intensive glucose control reduces morbidity and mortality after HSCT.
Biology of Blood and Marrow Transplantation
Psicologia: Ciência e Profissão, 2004
Resumo: O presente estudo, objetiva investigar a representação simbólica do câncer em profissiona... more Resumo: O presente estudo, objetiva investigar a representação simbólica do câncer em profissionais da área de saúde, pacientes, familiares e população em geral. Investigou-se, as imagens simbólicas que permeiam a atitude de cada um perante a convivência com o câncer por meio de questionário. Os resultados indicaram informações ricas de subjetividade, independentemente das diferenças demográficas. A imagética interfere na percepção, na reação e na distribuição do tempo que cada um determina para a escuta da singularidade daquele que adoece com o câncer e/ou com a contaminação simbólica. Na medida em que o fantasma da morte acompanha o paciente em seus movimentos, espera-se que este estudo possa contribuir para que se busquem formas de aprimoramento do tratamento do paciente com câncer. No entanto, para ajudar o outro, é preciso, primeiramente, redimensionar a própria formação imagética; afinal, nós, profissionais da área de saúde, também somos semeadores das imagens que se reproduzem ao longo dos séculos, apesar de toda a evolução científica. Palavras-Chave: Câncer, representação simbólica, qualidade de vida.
Cytokine, 2013
Fanconi anemia (FA) is a rare disease, autosomal recessive and X linked, which is clinically pron... more Fanconi anemia (FA) is a rare disease, autosomal recessive and X linked, which is clinically prone to development of hematological abnormalities and neoplasms, especially acute myeloid leukemia. In this work IL-10 and TGF-b levels were measured on FA patients' plasma since they are the regulatory cytokines of TNF-a and INF-c which had been described to be overexpressed in this genetic disease. Our results show increased IL-10 plasma levels in 25% of FA patients studied, but levels of TGF-b within the normal range. TNF-a and INF-c were also measured and found to be increased in 24% and 23% of FA patients, respectively. However, no inverse correlation was observed between augmented levels of IL-10 and TNF or IFN-c. Patients with elevated levels of TNF-a and INF-c presented bone marrow hypocellularity. IL-10 levels did not appear to be determinant for bone marrow cellularity. These data suggest that IL-10 is also a feature of Fanconi anemia pathophysiology.
Biology of Blood and Marrow Transplantation, 2016
We report on long-term survival in 157 patients with Fanconi anemia (FA) who survived 2 years or ... more We report on long-term survival in 157 patients with Fanconi anemia (FA) who survived 2 years or longer after their first transplant with a median follow-up of 9 years. Marrow failure (80%) was the most common indication for transplantation. There were 20 deaths beyond 2-years post-transplantation with 12 of the deaths occurring beyond 5 years after transplantation. Donor chimerism was available for 149 patients; n=112 (76%) reported >95%, n=27 (18%), 90-95%, and n=8 (5%), 20-89% donor chimerism. Two patients have <20% donor chimerism. The 10- and 15-year probabilities of survival were 90% and 79%, respectively. Results of multivariate analysis showed higher mortality risks for transplantations prior to 2003 (HR: 7.87, p=0.001), chronic GVHD (HR: 3.80, p=0.004) and squamous cell carcinoma, post-transplant (HR 38.17, p<0.0001). The predominant cause of late mortality was squamous cell carcinoma with an incidence of 8% and 14% at 10 and 15 years post-transplantation, respectively, and was more likely to occur in those with chronic GVHD. Other causes of late mortality included chronic GVHD, infection, graft failure, other cancers and hemorrhage. Although most patients are disease-free and functional long-term, our data support aggressive surveillance for long periods to identify those at risk for late mortality.
Haematologica, 2004
As some patients with Fanconi s anemia (FA) present excessive telomere shortening correlating wit... more As some patients with Fanconi s anemia (FA) present excessive telomere shortening correlating with poor outcome, we investigated whether human telomerase RNA component (hTERC) mutations also play a role in telomere shortening in 115 FA patients. Only one patient was heterozygous for the G58A polymorphism. No other mutation or deletion was found. We conclude that hTERC gene mutations do not contribute to telomere shortening in FA.
Revista Brasileira De Hematologia E Hemoterapia, 2010
Revista Brasileira de Hematologia e Hemoterapia, 2003
Page 1. 239 Pasquini R et al Rev. bras. hematol. hemoter. 2003;25(4):239-246 Relato de caso / Cas... more Page 1. 239 Pasquini R et al Rev. bras. hematol. hemoter. 2003;25(4):239-246 Relato de caso / Case report Carcinoma de células escamosas em língua pós-transplante de medula óssea por Anemia de Fanconi Squamous ...
Jornal de Pediatria, 1999
INTRODUCTION: The Imerslund-Gräsbeck syndrome is a rare hereditary autosomal recessive disease, c... more INTRODUCTION: The Imerslund-Gräsbeck syndrome is a rare hereditary autosomal recessive disease, characterized by the onset of megaloblastic anemia and asymptomatic proteinuria during the first 2 years of life. OBJECTIVE: To emphasize the importance of early detection of this disorder, due to high morbidity when not correctly treated, in addition to the necessity of screening and genetic counseling of the asymptomatic family members. METHODS: The authors report two patients, male and female, 8 and 10 years old, respectively. Their past history revealed anemia and multiple blood transfusions since their infancy. They evolved with pancytopenia during childhood and diagnosis of Severe Aplastic Anemia or Fanconi Syndrome was suspected. They were referred to the Bone Marrow Transplantation Section -HC- UFPR. RESULTS: Laboratory investigations revealed pancytopenia in peripheral blood. Bone marrow aspiration showed a marked megaloblastic erythropoiesis. Twenty-four-hour urine collection revealed proteinuria (3.0 and 5.8 g/dl respectively). Cytogenetic analysis was normal. Resolution of symptoms followed replacement therapy with parenteral vitamin B12. CONCLUSIONS: The presence of megaloblastic anemia in children should be followed by investigation of proteinuria, due to the existence of this rare disorder, that has a simple diagnosis and an effective treatment.
Revista Brasileira de Hematologia e Hemoterapia, 2008
Imatinib mesylate is currently the gold-standard therapy for patients with newly diagnosed Chroni... more Imatinib mesylate is currently the gold-standard therapy for patients with newly diagnosed Chronic Myelogenous Leukemia. From the clinical trials in 1998 to the IRIS study, which compared first line imatinib treatment with interferon and low dose ara-C, this drug has been consolidated in regards to its safety and efficacy. There are still some questions to answer. Which would be the best initial dose? Are there any patients who benefit more than others? What is the best approach to suboptimal response and resistance? The most important published clinical studies are reviewed in the current article and discussed from a Brazilian perspective.
Revista Brasileira de Hematologia e Hemoterapia, 2010
... Os pacientes em ambos os grupos eram comparáveis em relação à idade, sexo, estádio, isotipo d... more ... Os pacientes em ambos os grupos eram comparáveis em relação à idade, sexo, estádio, isotipo da proteína monoclonal e nível de β2-microglobulina. ... As características e fatores de risco dos pacientes eram semelhantes entre os quatro grupos. ...
Human Mutation, 2003
Fanconi anemia (FA) is a genetically heterogeneous autosomal recessive syndrome associated with c... more Fanconi anemia (FA) is a genetically heterogeneous autosomal recessive syndrome associated with chromosomal instability, hypersensitivity to DNA cross-linking agents, and predisposition to malignancy. The gene for FA complementation group G (FANCG) was the third FA gene to be cloned, and was found to be identical with human XRCC9, which maps to 9p13. The cDNA is predicted to encode a polypeptide of 622 amino acids, with no sequence similarities to any other known protein or motifs that could point to a molecular function for FANCG/XRCC9. We used single strand conformational polymorphism analysis (SSCP) to screen genomic DNA from a panel of 307 racially and ethnically diverse unrelated FA patients from the International Fanconi Anemia Registry (IFAR) for variants in FANCG. Twenty-seven abnormal SSCP patterns were found; 18 of these variants appear to be pathogenic mutations while nine are likely to be nonpathogenic polymorphisms. Direct sequencing of genomic DNA from seven FA-G probands with one mutant allele not detected in the SSCP study and three additional probands assigned to the FA-G complementation group by retroviral correction with FANCG resulted in the detection of nine additional pathogenic mutations and two common SNPs. Conditions for rapid screening for these mutations by DHPLC for use in a clinical laboratory setting were established. The most common FANCG mutations in the IFAR population were: IVS8-2A>G (seven Portuguese-Brazilian probands), IVS11+1G>C (seven French-Acadian probands), 1794_1803del10 (seven European probands), and IVS3+1G>C (five Korean or Japanese probands). Our data suggest that the Portuguese-Brazilian, French-Acadian, and Korean/Japanese mutations were likely to have been present in a founding member of each of these populations.
European Journal of Haematology, 2005
Fanconi anaemia (FA) is a rare genetic disease whose patients have a high predisposition to haema... more Fanconi anaemia (FA) is a rare genetic disease whose patients have a high predisposition to haematological abnormalities and cancer. Fas expression levels in peripheral blood lymphocytes samples of 73 FA patients were measured to verify if alterations in Fas expression could lead to predisposition/resistance to spontaneous or PHA induced apoptosis, as well as, to reflect some haematological features of this disease. The anti- and pro-apoptotic proteins Bcl-2 and Bax were also evaluated. FA patients samples could be divided into three different groups based on Fas expression: 20 samples had low, 32 normal and 21 increased Fas levels when compared to 41 control samples. No correlation was found between Fas and Bcl-2 expression but a good association was obtained with Bax, in the subgroup with increased Fas expression. The best correlation was seen between Bax expression and apoptosis. Out of the 15 samples with high Bax expression, 11 underwent apoptosis whereas only one out of seven samples with low levels of Bax displayed increased induced apoptosis. Most patients with normal haematological features expressed Fas within normal levels. It is difficult to establish, however, if Fas-expression is involved in the cause or is a consequence of the effects observed.
Brazilian Archives of Biology and Technology, 2005
ABSTRACT
Biology of Blood and Marrow Transplantation, 2014
Biology of Blood and Marrow Transplantation, 2007
The major complication associated with allogeneic hematopoietic stem cell transplantation (HSCT) ... more The major complication associated with allogeneic hematopoietic stem cell transplantation (HSCT) is graft-versus-host disease (GVHD), which is mediated by mature T cells in the donor transplant inoculum. Since donor T cells are also important in preventing graft failure, opportunistic infections and leukemia relapses after transplant, it would be highly advantageous to identify and separate T cells with GVHD-reactivity from those capable of mediating graft-versus-leukemia (GVL) responses. In this regard, TCR V repertoire analysis by CDR3-size spectratyping can be a powerful tool to characterize alloreactive T cell responses. As an initial test of this capability, we studied lethal GVHD in the B6 -Ͼ CXB-2 MHC-matched, minor histocompatibility antigen disparate murine model to determine the correlation between the in vitro detected alloreactive B6 CD8 ϩ T cell responses and their actual involvement in the development of GVHD upon transfer to lethally irradiated CXB-2 recipients. Previous methods to obtain the V repertoire from responding mixed lymphocyte reactions (MLR) required expansion of the alloreactive cells in secondary stimulation cultures, for a total of 20 days. To significantly shorten this incubation period, in order to eventually adapt this technique for clinical use, we used the fluorescent-based intracellular dye CFSE to label responding cells in a 4-day MLR and fluorescent cell sorting to isolate the divided alloreactive cells that could then be processed for V spectratype analysis. In a comparative study of the predictive value of the V repertoire detected in traditional MLR versus CFSE/MLR, the results indicated that the former approach correlated with 78% of the V CDR3-size skewing patterns observed in mice with B6 -Ͼ CXB-2 GVHD, and the CFSE/MLR approach corresponded to 65% of those skewed V families found in vivo. These results suggest that in vitro spectratyping analysis, and particularly the use of CFSE to predict in vivo reactivity, may be a useful technique in eventually guiding the manipulation of the donor T cell inoculum in order to improve the outcome of HSCT.
Biology of Blood and Marrow Transplantation, 2007
pts engrafted at a md of 22 (11-41) days. 6 pts lost complete chimerism during follow up with pro... more pts engrafted at a md of 22 (11-41) days. 6 pts lost complete chimerism during follow up with proven relapse at a md of 401 days in four. At a md follow up of 23 (4-79) months probability of overall (OS) vs eventfree survival (EFS) was 74% vs 50% for the whole cohort. A trend towards better survival was seen in recipients of related compared to unrelated transplants (100 vs 64 %, pϭ0.06) and in those with less advanced disease (90 vs 68 % in Dupriez 0 vs 1ϩ2, pϭ0.26). However the only significant predictor for OS by log rank test was the pretransplant CCI: 85 vs 34 % in pts with CCI 0 to 2 vs 3 to 6 (pϭ0.01). Probability of EFS at day 480 was significantly less in pts with abnormal karyoptype compared to those with no detectable anomalies (17 vs 61%, pϭ0.01). Five pts with decreasing chimerism received immunotherapy (3 DLI, 2 PBSC) with return to complete chimerism in three and postDLI aplasia in one.-Conclusions: Our results support the use of related and unrelated alloHCT as a curative treatment option in MMM. Outcome is better in pts without disease associated and disease independent comorbidity. For pts with abnormal karyotype there is a considerable risk of relapse/reversal post transplant. Especially in risk pts chimerism therefore should be monitored closely as disease recurrence may respond to immunotherapy.
Biology of Blood and Marrow Transplantation, 2007
We have demonstrated that patient derived myeloma cells fused with autologous dendritic cells (DC... more We have demonstrated that patient derived myeloma cells fused with autologous dendritic cells (DCs) potently stimulate anti-tumor immunity in vitro. We are conducting phase I clinical trials in which patients with myeloma undergo serial vaccination with DC/ myeloma fusions alone or in conjunction with stem cell transplantation. To date, 18 patients have been enrolled (11-vaccine alone, 7 vaccine following autologous stem cell transplant). To generate mature DCs, adherent mononuclear cells were isolated from a leukapharesis collection and cultured with GM-CSF, IL-4, and TNF␣ . The mean yield and viability of the DC preparations was 1.5 ϫ 10 8 cells and 88%, respectively. Patient derived myeloma cells were isolated from bone marrow aspirates and were quantified by the expression of CD38 and/or CD138. The mean yield and viability of the myeloma cell collections was 7.3 ϫ 10 7 cells and 89%, respectively. Fusion cells were generated by coculture of DCs with myeloma cells in the presence of 50% polyethylene glycol. The mean fusion efficiency was 40% as determined by the percentage of cells that co-expressed unique DC and myeloma antigens. In contrast to myeloma cells, the DC and fusion cell preparations prominently stimulated allogeneic T cell proliferation in vitro. To date, 13 patients have completed vaccination at a dose of 1-5 ϫ 10 6 fusion cells. GM-CSF (100 g) was administered subcutaneously on the day of vaccination and for 3 days thereafter. Adverse events judged to be potentially vaccine related have included vaccine injection site reactions, edema, rash, fever, chills, fatigue, muscle aches, pruritis, and diarrhea. One patient with a history of prior deep venous thrombosis (DVT) developed a DVT and pulmonary embolus of uncertain relation to the vaccine. To date, a majority of evaluable patients demonstrated evidence of vaccine induced anti-myeloma immunity as demonstrated by at least 2 fold increase in IFN␥ expression by CD4 and/or CD8 T cells in response to ex vivo exposure to autologous tumor lysate. Of patients undergoing vaccine therapy alone, 5 patients demonstrated stabilization of the myeloma paraprotein for 2-6 months following initiation of vaccination. Of 3 patients completing posttransplant vaccination, 2 patients demonstrated resolution of the persisting myeloma protein post-transplant and 1 patient demonstrated a transient increase followed by a decline in paraprotein levels post-transplant.
Biology of Blood and Marrow Transplantation, 2007
sults There was no significant difference between the 3 groups in the occurrence of FN or documen... more sults There was no significant difference between the 3 groups in the occurrence of FN or documented infection. However, hyperglycemia was significantly associated with organ dysfunction and aGVHD. OS was better and TRM was less in group1 compared with group2 and group3. Conclusion Degrees of hyperglycemia during neutropenia was associated with an increased risk of organ dysfunction and aGVHD, which further led to higher TRM and lower OS. These results support the possibility that intensive glucose control reduces morbidity and mortality after HSCT.