RESUMO
This phase I trial reports the safety and activity of BPX101, a second-generation antigen-targeted autologous antigen presenting cell (APC) vaccine in men with metastatic castration-resistant prostate cancer (mCRPC). To manufacture BPX101, APCs collected in a single leukapheresis were transduced with adenoviral vector Ad5f35 encoding inducible human (ih)-CD40, followed by incubation with protein PA001, which contains the extracellular domain of human prostate-specific membrane antigen. The ih-CD40 represents a modified chimeric version of the dendritic cell (DC) co-stimulatory molecule, CD40, which responds to a bioinert membrane-permeable activating dimerizer drug, rimiducid (AP1903), permitting temporally controlled, lymphoid-localized, DC-specific activation. Eighteen men with progressive mCRPC following ≤1 prior chemotherapy regimen were enrolled to evaluate three doses of BPX101 (4 × 106, 12.5 × 106 and 25 × 106 cells) administered intradermally every 2-4 weeks followed by rimiducid (0.4 mg/kg) intravenous (IV) infusion 24 h after each BPX101 dose. There were no dose-limiting toxicities. Immune upregulation as well as anti-tumor activity was observed with PSA declines, objective tumor regressions and robust efficacy of post-trial therapy. This novel antigen-targeted and in vivo activated immunotherapy platform may warrant further development as monotherapy and as a component of rational combinations.
Assuntos
Antígenos CD40/metabolismo , Vacinas Anticâncer/imunologia , Células Dendríticas/imunologia , Neoplasias da Próstata/imunologia , Idoso , Vacinas Anticâncer/uso terapêutico , Estudos de Coortes , Humanos , MasculinoRESUMO
BACKGROUND: Poor engraftment due to low cell doses restricts the usefulness of umbilical-cord-blood transplantation. We hypothesized that engraftment would be improved by transplanting cord blood that was expanded ex vivo with mesenchymal stromal cells. METHODS: We studied engraftment results in 31 adults with hematologic cancers who received transplants of 2 cord-blood units, 1 of which contained cord blood that was expanded ex vivo in cocultures with allogeneic mesenchymal stromal cells. The results in these patients were compared with those in 80 historical controls who received 2 units of unmanipulated cord blood. RESULTS: Coculture with mesenchymal stromal cells led to an expansion of total nucleated cells by a median factor of 12.2 and of CD34+ cells by a median factor of 30.1. With transplantation of 1 unit each of expanded and unmanipulated cord blood, patients received a median of 8.34×10(7) total nucleated cells per kilogram of body weight and 1.81×10(6) CD34+ cells per kilogram--doses higher than in our previous transplantations of 2 units of unmanipulated cord blood. In patients in whom engraftment occurred, the median time to neutrophil engraftment was 15 days in the recipients of expanded cord blood, as compared with 24 days in controls who received unmanipulated cord blood only (P<0.001); the median time to platelet engraftment was 42 days and 49 days, respectively (P=0.03). On day 26, the cumulative incidence of neutrophil engraftment was 88% with expansion versus 53% without expansion (P<0.001); on day 60, the cumulative incidence of platelet engraftment was 71% and 31%, respectively (P<0.001). CONCLUSIONS: Transplantation of cord-blood cells expanded with mesenchymal stromal cells appeared to be safe and effective. Expanded cord blood in combination with unmanipulated cord blood significantly improved engraftment, as compared with unmanipulated cord blood only. (Funded by the National Cancer Institute and others; ClinicalTrials.gov number, NCT00498316.).
Assuntos
Transplante de Células-Tronco de Sangue do Cordão Umbilical , Neoplasias Hematológicas/terapia , Transplante de Células-Tronco Mesenquimais , Adolescente , Adulto , Contagem de Células Sanguíneas , Plaquetas , Causas de Morte , Técnicas de Cultura de Células , Facilitação Imunológica de Enxerto , Doença Enxerto-Hospedeiro , Neoplasias Hematológicas/mortalidade , Humanos , Células-Tronco Mesenquimais , Pessoa de Meia-Idade , Neutrófilos , Quimeras de Transplante , Transplante Homólogo , Adulto JovemRESUMO
BACKGROUND AIMS: Wide acceptance of the colony-forming unit (CFU) assay as a reliable potency test for stem cell products is hindered by poor inter-laboratory reproducibility. The goal of this study was to ascertain current laboratory practices for performing the CFU assay with an eye towards identifying practices that could be standardized to improve overall reproducibility. METHODS: A survey to evaluate current laboratory practices for performing CFU assays was designed and internationally distributed. RESULTS: There were 105 respondents to the survey, of whom 68% performed CFU assays. Most survey recipients specified that an automated rather than a manual cell count was performed on pre-diluted aliquots of stem cell products. Viability testing methods employed various stains, and when multiple sites used the same viability stain, the methods differed. Cell phenotype used to prepare working cell suspensions for inoculating the CFU assay differed among sites. Most respondents scored CFU assays at 14-16 days of incubation, but culture plates were read with various microscopes. Of 57 respondents, 42% had not performed a validation study or established assay linearity. Only 63% of laboratories had criteria for determining if a plate was overgrown with colonies. CONCLUSIONS: Survey results revealed inconsistent inter-laboratory practices for performing the CFU assay. The relatively low number of centers with validated CFU assays raises concerns about assay accuracy and emphasizes a need to establish central standards. The survey results shed light on numerous steps of the methodology that could be targeted for standardization across laboratories.
Assuntos
Terapia Baseada em Transplante de Células e Tecidos , Ensaio de Unidades Formadoras de Colônias/normas , Células-Tronco Hematopoéticas , Células-Tronco/citologia , Antígenos CD34/metabolismo , Transplante de Células-Tronco Hematopoéticas , Humanos , Padrões de Referência , Reprodutibilidade dos TestesRESUMO
Alternative donor stem cell transplantation from cord blood or haploidentical peripheral blood donors is increasingly being used for patients who lack a matched related or unrelated donor. A higher nonrelapse mortality (NRM) rate has been noted with these 2 types of transplants, primarily because of infectious complications. Here, we hypothesized that the time to lymphocyte recovery (absolute lymphocyte count [ALC] of ≥1000/µL for the first 3 consecutive days) after transplant correlates with outcomes. We retrospectively analyzed 65 consecutive patients treated at our institution with cord blood (n = 37) and haploidentical (n = 28) transplantation with myeloablative fludarabine, melphalan, and thiotepa conditioning. Patients with lymphocyte recovery at day 60 posttransplant were more likely to survive long term than those without lymphocyte recovery. In multivariate analysis, ALC recovery was the only independent prognostic factor associated with mortality; patients without ALC recovery were 10.5 times (95% confidence interval [CI]: 4.3-25.4) more likely to die than those with ALC recovery (P < .0001). This difference appeared to be related to NRM (hazard ratio [HR] = 0.1, 95% CI: 0.02-0.6, P = .008), whereas ALC recovery did not influence the rate of disease relapse. These results suggest that ALC recovery is an important prognostic indicator for patients treated with cord blood and T cell-depleted peripheral haploidentical transplants.
Assuntos
Transplante de Células-Tronco de Sangue do Cordão Umbilical/métodos , Transplante de Células-Tronco Hematopoéticas/métodos , Linfócitos/imunologia , Doença Aguda , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Haploidia , Haplótipos , Humanos , Leucemia/sangue , Leucemia/imunologia , Leucemia/cirurgia , Contagem de Linfócitos , Linfócitos/citologia , Linfoma/sangue , Linfoma/imunologia , Linfoma/cirurgia , Masculino , Pessoa de Meia-Idade , Taxa de Sobrevida , Linfócitos T/citologia , Linfócitos T/imunologia , Condicionamento Pré-Transplante , Resultado do Tratamento , Adulto JovemRESUMO
The following commentary was developed by the National Marrow Donor Program Cord Blood Advisory Group and is intended to provide an overview of umbilical cord blood (UCB) processing, summarize the current state of potency assays used to characterize UCB, and define limitations of the assays and future needs of the cord blood banking and transplant community. The UCB banking industry is eager to participate in the development of standardized assays to uniformly characterize cellular therapy products that are manufactured in a variety of ways. This paper describes the desired qualities of these assays and how the industry proposes to co-operate with developers to bring relevant assays to market. To that end, the National Marrow Donor Program (NMDP) Cord Blood Bank Network is available to serve as a resource for UCB testing material, research and development consulting, and product/assay testing in an accredited UCB manufacturing environment.
Assuntos
Bioensaio/métodos , Bioensaio/normas , Sangue Fetal , Bancos de Sangue/normas , Transplante de Células/métodos , Humanos , Reprodutibilidade dos TestesRESUMO
BACKGROUND: Most hematopoietic progenitor cell (HPC) products are infused or processed shortly after collection, but in some cases this may be delayed for up to 48 hours. A number of variables such as temperature and cell concentration are of critical importance for the integrity of HPCs during this time. STUDY DESIGN AND METHODS: We evaluated critical variables using cord blood HPC units that were divided equally and stored at 4 °C versus room temperature (RT) for up to 96 hours. Total nucleated cell (TNC) and mononuclear cell (MNC) counts, viable CD34+ cell counts, and CD45+ cell viability as well as colony-forming unit-granulocyte-macrophage (CFU-GM) present over time at each temperature were determined. RESULTS: Overall, the data indicate that with the exception of viable CD34+ cells, there was a significant decrease in each variable measured for 72 to 96 hours and, with the exception of viable CD34+ cells and CFU-GM, the reductions were significantly greater in RT units than 4 °C units. There was an increase in viable CD34+ count for units where TNC count was greater than 8.5 × 10(9) /L, compared with units where TNC count was less than 8.5 × 10(9) /L, that was different for each storage temperature. CONCLUSIONS: Cord blood HPC collections maintained at 4 °C retained higher TNC counts, MNC counts, and CD45+ cell viability over a 72- to 96-hour storage period.
Assuntos
Preservação de Sangue/métodos , Células Progenitoras de Granulócitos e Macrófagos/citologia , Células-Tronco Hematopoéticas/citologia , Antígenos CD34/metabolismo , Sangue Fetal/citologia , TemperaturaRESUMO
BACKGROUND: The purpose of this study was to perform a detailed analysis of the charges associated with chemomobilization and remobilization of autologous hematopoietic stem cells (HSCs) and to quantify medical costs and resource utilization associated with these procedures. STUDY DESIGN AND METHODS: Patients with lymphoma underwent chemomobilization with ifosfamide and etoposide with or without rituximab (IE ± R). Patients with multiple myeloma (MM) received a modified hyperfractionated cyclophosphamide, vincristine, doxorubicin, dexamethasone (hyper-CVAD) regimen after failing to mobilize with growth factors only. RESULTS: Between January 2004 and October 2006, 98 patients with lymphoma underwent HSC mobilization with IE ± R. Mobilization with IE ± R was effective, with 90.8% of patients collecting at least 2 × 10(6) CD34+ cells/kg. The total charges for treatment were $27,996 and $37,667 for patients mobilized with IE and IE + R, respectively. Hospital readmission for complications occurred in 26.5% of patients, resulting in additional charges of $10,356. The preapheresis procedure charge was estimated to be $2522, the charge for a 2-day apheresis session was $5160, and the postapheresis phase resulted in charges of $8040. Our analysis determined that reducing apheresis by 1 day has the potential to save $6600. We also performed a retrospective analysis of 16 patients with MM remobilized with a modified hyper-CVAD regimen. Remobilization was successful, with 87.5% of patients. Our analysis determined that mobilization, preapheresis, apheresis, and postapheresis phase charges were $24,968, $2522, $6158, and $12,060, respectively. CONCLUSIONS: Optimization of HSC mobilization regimens to reduce failure rates would not only benefit patients but also reduce the overall medical costs.
Assuntos
Mobilização de Células-Tronco Hematopoéticas/métodos , Linfoma/terapia , Mieloma Múltiplo/terapia , Transplante de Células-Tronco de Sangue Periférico/métodos , Transplante Autólogo/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Monoclonais Murinos/uso terapêutico , Antineoplásicos/uso terapêutico , Ciclofosfamida/uso terapêutico , Dexametasona/uso terapêutico , Doxorrubicina/uso terapêutico , Etoposídeo/uso terapêutico , Feminino , Humanos , Ifosfamida/uso terapêutico , Linfoma/tratamento farmacológico , Masculino , Pessoa de Meia-Idade , Mieloma Múltiplo/tratamento farmacológico , Rituximab , Adulto JovemRESUMO
BACKGROUND: Interlaboratory scoring performances were determined using a traditional 14-day colony-forming unit (CFU) assay and a new 7-day CFU assay. STUDY DESIGN AND METHODS: Digital images of colonies were utilized to train personnel at each site. A central laboratory inoculated methylcellulose with progenitors and sent the samples by overnight courier to participating labs for plating. RESULTS: Colony counts from two digital images showed greater variability by novice counters (coefficients of variation [CV], 18.5 and 23.0%; n = 8) than for experienced staff (CV, 7.3 and 4.8%; n = 5). CFU assays plated immediately, 24 and 48 hours after methylcellulose inoculation displayed 39.5 CFU, 37.1 ± 10.6 (CV, 28%) and 34.8 ± 8.5 (CV, 24%) colonies for the 7-day assay and 39.5 CFU, 39.1 ± 9.9 (CV, 25%) and 37.1 ± 10.6 (CV, 28%) colonies for the 14-day assay, respectively. Overall, no significant differences in colony counts were noted between assays (p = 0.68). Also, no differences in CFU counts were seen when assays were set up immediately, 24 and 48 hours after methylcellulose inoculation (14-day p = 0.695; 7-day p = 0.632). CONCLUSION: Total CFUs obtained in 7- and 14-day CFU assays are comparable and show similar levels of interlaboratory variability. The major source of this variability is due to differences in how CFU plates are scored by individuals at different sites. UCB progenitor cells can be maintained in methylcellulose-based media at room temperature for up to 48 hours prior to transport without a significant loss in CFUs.
Assuntos
Ensaio de Unidades Formadoras de Colônias/métodos , Humanos , Fatores de TempoRESUMO
Lenalidomide is an agent that has shown great activity in patients with multiple myeloma (MM). However, studies have suggested that this drug negatively affects subsequent stem cell collection. To investigate whether lenalidomide impairs stem cell mobilization and collection, we reviewed data for patients with MM who underwent mobilization with filgrastim. Predictors of mobilization failure were evaluated using logistic regression analysis. In 26 (9%) of 302 myeloma patients, stem cell mobilization failed. Mobilization failed in 25% of patients who had previously received lenalidomide, compared with 4% of patients who had not received lenalidomide (P < .001). In a multivariate analysis, prior lenalidomide use (odds ratio: 5.9; 95% confidence interval [CI]: 2.4-14.3) and mobilization more than 1 year after diagnosis (odds ratio: 4.6; 95% CI: 1.9-11.1) were significantly associated with failed mobilization. Twenty-one of 26 patients in whom mobilization with filgrastim failed underwent remobilization with chemotherapy and filgrastim; in 18 (86%) of these 21 patients, stem cells were successfully mobilized and collected. In patients with multiple myeloma, prior lenalidomide therapy is associated with failure of stem cell mobilization with filgrastim. Remobilization with chemotherapy and filgrastim is usually successful in these patients.
Assuntos
Antineoplásicos/efeitos adversos , Medula Óssea/efeitos dos fármacos , Fator Estimulador de Colônias de Granulócitos/antagonistas & inibidores , Mobilização de Células-Tronco Hematopoéticas/métodos , Mieloma Múltiplo/sangue , Transplante de Células-Tronco de Sangue Periférico , Talidomida/análogos & derivados , Adulto , Idoso , Inibidores da Angiogênese/administração & dosagem , Inibidores da Angiogênese/efeitos adversos , Inibidores da Angiogênese/uso terapêutico , Antineoplásicos/administração & dosagem , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Benzilaminas , Terapia Combinada , Ciclamos , Ciclofosfamida/administração & dosagem , Ciclofosfamida/farmacologia , Dexametasona/administração & dosagem , Doxorrubicina/administração & dosagem , Etoposídeo/administração & dosagem , Etoposídeo/farmacologia , Feminino , Filgrastim , Fator Estimulador de Colônias de Granulócitos/administração & dosagem , Fator Estimulador de Colônias de Granulócitos/farmacologia , Compostos Heterocíclicos/administração & dosagem , Compostos Heterocíclicos/farmacologia , Humanos , Ifosfamida/administração & dosagem , Ifosfamida/farmacologia , Lenalidomida , Leucaférese , Masculino , Pessoa de Meia-Idade , Mieloma Múltiplo/tratamento farmacológico , Mieloma Múltiplo/radioterapia , Mieloma Múltiplo/cirurgia , Proteínas Recombinantes , Estudos Retrospectivos , Fatores de Risco , Talidomida/administração & dosagem , Talidomida/efeitos adversos , Talidomida/uso terapêutico , Transplante Autólogo , Falha de Tratamento , Vincristina/administração & dosagemRESUMO
The purpose of this retrospective study was to determine the incidence and predictive factors if any, of mobilization failure in lymphoma patients referred for autologous stem cell transplantation. A total of 588 lymphoma patients were referred for transplant consultation from January 2003 to December 2004. Predictors of mobilization failure were evaluated using logistic regression analysis including diagnosis, mobilization regimen, age, sex, type and number of prior chemotherapies, bone marrow cellularity, platelet count, white count, prior bone marrow involvement with malignancy, and prior radiation therapy. Two hundred and six patients were eligible for transplantation and underwent stem cell mobilization. Twenty-nine (14%) patients failed to mobilize adequate stem cells after the first attempt. For the entire group age (>or=60 versus <60 years), diagnosis (Hodgkin's versus non-Hodgkin's lymphoma), use of cytokines alone, platelet count <150 x 10(9)/L, and bone marrow cellularity <30% were significant predictors for mobilization failure on univariate analysis. In view of small number of patients multivariate analysis was not possible. However, a low platelet count (150 x 10(9)/L) was the only significant predictor when the analysis was restricted to non-Hodgkin's lymphoma patients who were mobilized with chemotherapy. Mobilization failure rates are higher in patients with non-Hodgkin's lymphoma compared with those with Hodgkin's lymphoma. In the subset of patients who undergo chemomobilization for non-Hodgkin's lymphoma platelet count at the time of mobilization is a predictor of mobilization failure.
Assuntos
Mobilização de Células-Tronco Hematopoéticas/métodos , Linfoma/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Transplante de Células-Tronco Hematopoéticas/métodos , Humanos , Linfoma/cirurgia , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/patologia , Recidiva Local de Neoplasia/cirurgia , Recidiva , Análise de Regressão , Estudos Retrospectivos , Fatores de Risco , Transplante Autólogo , Adulto JovemRESUMO
The long-term outcome of graft failure after allogeneic stem cell transplantation (SCT) has not been well described. To fill this knowledge gap we performed a retrospective analysis of patients with graft failure over a 10-year time period in a single institution. Cases were included for analysis if they had failed to achieve an absolute neutrophil count (ANC) of 500/microL or more by 28 days post-SCT or 42 days after cord blood transplantation (primary graft failure); had a decrease in their ANC to <500/mL for 3 consecutive days after having achieved neutrophil engraftment (secondary graft failure); or failed to have evidence of at least 5% or more donor cell engraftment (primary graft failure with autologous reconstitution). Among 1726 patients who underwent allografts from January 1, 1990, through December 31, 2000, we identified 68 patients with graft failure. The 1-, 2-, and 5-year overall survival (OS) for all patients was 31%, 24%, and 15%. A diagnosis of acute leukemia was a significant predictor for poor survival on multivariate analysis. We conclude that graft failure is an uncommon complication postallogeneic SCT, and is associated with poor outcomes. Collection of autologous stem cells prior to high-risk allografting can salvage a fraction of patients and lead to prolonged survivals.
Assuntos
Rejeição de Enxerto/etiologia , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Feminino , Seguimentos , Rejeição de Enxerto/diagnóstico , Transplante de Células-Tronco Hematopoéticas/mortalidade , Humanos , Contagem de Leucócitos , Masculino , Pessoa de Meia-Idade , Neutrófilos/citologia , Prognóstico , Estudos Retrospectivos , Análise de Sobrevida , Transplante Autólogo , Transplante HomólogoRESUMO
The Elutra biomedical device allows semi-automatic enrichment of monocytes by elutriation, using a single-use, closed and cGMP compliant tubing set, in a cost effective way. The procedure has been validated using fresh apheresis products from nonmobilized donors. We here evaluated the possibility of using Elutra to enrich monocytes from frozen/thawed apheresis products collected from mobilized healthy donors. Frozen apheresis products from 6 G CSF mobilized donors were thawed and used in 16 elutriation procedures. We compared the recovery and purity of enriched monocytes using different buffer compositions and elutriation profiles. Elutriated monocytes were cultured to generate mature dendritic cells (DCs). Depending in part of the initial granulocyte contamination in the apheresis product, the use of Desoxyribo Nuclease (DNAse) to avoid aggregation, was needed through only the initial steps or throughout the elutriation process. The average monocyte recovery was 85+/-31%. The average purity was 73+/-9%. The recovery of mature DC at d8 of culture was 20+/-6% of the input monocyte numbers. We conclude that Elutra allows the purification of monocytes from thawed mobilized apheresis. It requires no pre-processing of the cell product before elutriation, and allows the generation of phenotypically mature DC in quantities that are compatible with a clinical use.
Assuntos
Criopreservação/métodos , Leucaférese , Monócitos/citologia , Antígenos CD/análise , Contagem de Células , Diferenciação Celular/efeitos dos fármacos , Separação Celular/instrumentação , Separação Celular/métodos , Células Dendríticas/química , Células Dendríticas/citologia , Fator Estimulador de Colônias de Granulócitos/farmacologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Humanos , Interleucina-4/farmacologia , Monócitos/química , Monócitos/efeitos dos fármacos , Neutrófilos/citologia , Proteínas RecombinantesAssuntos
Transplante de Células-Tronco de Sangue do Cordão Umbilical , Animais , Bancos de Sangue/tendências , Transplante de Células-Tronco de Sangue do Cordão Umbilical/métodos , Transplante de Células-Tronco de Sangue do Cordão Umbilical/tendências , Seleção do Doador , Sangue Fetal , Genótipo , Sobrevivência de Enxerto/imunologia , Infecções por HIV/terapia , Hematologia/tendências , Histocompatibilidade , Humanos , Receptores KIR/imunologia , Medicina Regenerativa/tendências , Talassemia/terapia , Condicionamento Pré-TransplanteRESUMO
Rituximab, an anti-CD20 human-mouse chimeric monoclonal antibody has been shown to improve response rates when it is combined with standard salvage chemotherapy in patients with relapsed or refractory intermediate-grade B-cell non-Hodgkin's lymphoma. A vast majority of these patients subsequently undergo high-dose therapy followed by stem cell transplantation. However, the impact of rituximab on stem cell mobilization kinetics is not well characterized. The purpose of this study was to study the effect of high-dose rituximab given with chemotherapy on stem cell mobilization in patients with intermediate-grade B-cell non-Hodgkin's lymphoma. Thirty-six patients received ifosfamide, etoposide, and rituximab followed by filgrastim for stem cell mobilization. The chemotherapy regimen was well tolerated. Thirty-four of 36 patients (94%) were able to mobilize at least 2 x 10(6) CD34+ cells/kg body weight after a median of 2 apheresis procedures. The median CD34+ cell dose collected per kilogram of recipient body weight was 6.5 x 10(6) (range, 4.65-31.15). All patients who subsequently underwent high-dose chemotherapy and stem cell transplantation experienced sustained engraftment. In conclusion, high-dose rituximab given during stem cell mobilization does not negatively affect stem cell mobilization kinetics.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Antineoplásicos/uso terapêutico , Mobilização de Células-Tronco Hematopoéticas/métodos , Linfoma não Hodgkin/tratamento farmacológico , Linfoma não Hodgkin/terapia , Células-Tronco/efeitos dos fármacos , Adulto , Idoso , Anticorpos Monoclonais Murinos , Antígenos CD20/biossíntese , Antígenos CD34/biossíntese , Etoposídeo/administração & dosagem , Feminino , Filgrastim , Fator Estimulador de Colônias de Granulócitos/administração & dosagem , Humanos , Ifosfamida/administração & dosagem , Fatores Imunológicos/administração & dosagem , Cinética , Masculino , Pessoa de Meia-Idade , Proteínas Recombinantes , Rituximab , Transplante de Células-Tronco/métodos , Células-Tronco/citologiaRESUMO
Induction of donor type chimerism in mildly prepared hosts without graft-versus-host disease (GvHD) is a most desirable goal in bone morrow transplantation. We have recently demonstrated in a mouse model that donor veto cytotoxic T lymphocytes (CTLs) can facilitate the induction of donor type chimerism in sublethally irradiated recipients without causing GvHD if they are effectively depleted of alloreactivity against host cells by means of stimulation against a third party. We extend this approach to human cells, by preparing CTLs in two major steps: primary culture in the absence of interleukin 2, leading to death by neglect of antihost clones, and addition of interleukin 2 and subsequent dilution of antihost clones as a consequence of the expansion of the anti-third-party clones. CTLs prepared in this way specifically suppress host cytotoxic T cells directed against antigens of the donor, but not against fourth-party antigens, as demonstrated in a standard (51)Cr release assay. We conclude that human anti-third-party CTLs afford a new source of veto cells that are depleted of potential graft-versus-host-reactive clones. The cells generated by this approach could potentially be used to facilitate engraftment of allogeneic hematopoietic stem cells.
Assuntos
Transplante de Medula Óssea/imunologia , Doença Enxerto-Hospedeiro/prevenção & controle , Depleção Linfocítica , Linfócitos T Citotóxicos/imunologia , Animais , Células Clonais , Doença Enxerto-Hospedeiro/imunologia , Humanos , Tolerância Imunológica/imunologia , Ativação Linfocitária/imunologia , Teste de Cultura Mista de Linfócitos , Depleção Linfocítica/métodos , Doadores de TecidosRESUMO
OBJECTIVE: To investigate the graft-vs-leukemia effect of allogeneic stem cell transplantation after a nonablative conditioning regimen as treatment for patients with chronic lymphocytic leukemia. PATIENTS AND METHODS: Patients were eligible to treatment if they were refractory or recurred after a prior response to fludarabine. Seventeen patients were treated. All patients received a preparative regimen of fludarabine (30 mg/m(2) daily for 3 days) and intravenous cyclophosphamide (750 mg/m(2) daily for 3 days). Ten patients received rituximab in addition to chemotherapy. The median time from diagnosis to transplant was 67 months. Nine of 17 patients had refractory disease. RESULTS: All patients had engraftment of donor cells. Eleven (65%) did not require platelet transfusions. Ten patients with persistent disease underwent immunomanipulation to augment GVL effects including immunosuppression withdrawal and donor lymphocyte infusion with or without rituximab treatment. Seven of these 10 patients had a complete response and 2 had a partial response; 8 of these 9 responders had received rituximab with their immunomanipulation process. The final response was complete remission in 12 and partial remission in 4 patients for an overall response rate of 94%. Overall survival was 100% for patients who received the combined chemo-rituximab conditioning regimen, vs 14% for those who received chemotherapy alone (p=0.03). CONCLUSION: Our results indicate that a pronounced GVL effect occurs after nonmyeloablative allogeneic hematopoietic transplantation for advanced CLL. This activity might be facilitated by rituximab. Prospective controlled trials are needed to define the role of nonablative allogeneic hematopoietic transplantation for treatment of this disease.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Transplante de Células-Tronco Hematopoéticas , Leucemia Linfocítica Crônica de Células B/terapia , Adulto , Idoso , Anticorpos Monoclonais Murinos , Feminino , Doença Enxerto-Hospedeiro/etiologia , Humanos , Tolerância Imunológica/efeitos dos fármacos , Leucemia Linfocítica Crônica de Células B/imunologia , Leucemia Linfocítica Crônica de Células B/mortalidade , Masculino , Pessoa de Meia-Idade , Rituximab , Transplante HomólogoRESUMO
High-dose myeloablative therapy with allogeneic hematopoietic transplantation is an effective treatment for hematologic malignancies, but this approach is associated with a high risk of complications. The use of relatively nontoxic, nonmyeloablative, or reduced-intensity preparative regimens still allows engraftment and the generation of graft-vs-malignancy effects, is potentially curative for susceptible malignancies, and reduces the risk of treatment-related morbidity. Two general strategies along these lines have emerged, based on the use of (1) immunosuppressive chemotherapeutic drugs, usually a purine analog in combination with an alkylating agent, and (2) low-dose total body irradiation, alone or in combination with fludarabine (Fludara).
Assuntos
Transplante de Medula Óssea , Transplante de Células-Tronco de Sangue Periférico , Antineoplásicos Alquilantes/uso terapêutico , Efeito Enxerto vs Tumor , Humanos , Imunossupressores/uso terapêutico , Neoplasias/terapia , Transplante Homólogo , Irradiação Corporal TotalRESUMO
BACKGROUND: Tumor suppressor gene TUSC2/FUS1 (TUSC2) is frequently inactivated early in lung cancer development. TUSC2 mediates apoptosis in cancer cells but not normal cells by upregulation of the intrinsic apoptotic pathway. No drug strategies currently exist targeting loss-of-function genetic abnormalities. We report the first in-human systemic gene therapy clinical trial of tumor suppressor gene TUSC2. METHODS: Patients with recurrent and/or metastatic lung cancer previously treated with platinum-based chemotherapy were treated with escalating doses of intravenous N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTAP):cholesterol nanoparticles encapsulating a TUSC2 expression plasmid (DOTAP:chol-TUSC2) every 3 weeks. RESULTS: Thirty-one patients were treated at 6 dose levels (range 0.01 to 0.09 milligrams per kilogram). The MTD was determined to be 0.06 mg/kg. Five patients achieved stable disease (2.6-10.8 months, including 2 minor responses). One patient had a metabolic response on positron emission tomography (PET) imaging. RT-PCR analysis detected TUSC2 plasmid expression in 7 of 8 post-treatment tumor specimens but not in pretreatment specimens and peripheral blood lymphocyte controls. Proximity ligation assay, performed on paired biopsies from 3 patients, demonstrated low background TUSC2 protein staining in pretreatment tissues compared with intense (10-25 fold increase) TUSC2 protein staining in post-treatment tissues. RT-PCR gene expression profiling analysis of apoptotic pathway genes in two patients with high post-treatment levels of TUSC2 mRNA and protein showed significant post-treatment changes in the intrinsic apoptotic pathway. Twenty-nine genes of the 82 tested in the apoptosis array were identified by Igenuity Pathway Analysis to be significantly altered post-treatment in both patients (Pearson correlation coefficient 0.519; p<0.01). CONCLUSIONS: DOTAP:chol-TUSC2 can be safely administered intravenously in lung cancer patients and results in uptake of the gene by human primary and metastatic tumors, transgene and gene product expression, specific alterations in TUSC2-regulated pathways, and anti-tumor effects (to our knowledge for the first time for systemic DOTAP:cholesterol nanoparticle gene therapy). TRIAL REGISTRATION: ClinicalTrials.gov NCT00059605.
Assuntos
Técnicas de Transferência de Genes , Terapia Genética/métodos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/genética , Nanopartículas/uso terapêutico , Proteínas Supressoras de Tumor/uso terapêutico , Apoptose/efeitos dos fármacos , Apoptose/genética , Relação Dose-Resposta a Droga , Ácidos Graxos Monoinsaturados/administração & dosagem , Perfilação da Expressão Gênica , Humanos , Dose Máxima Tolerável , Nanopartículas/administração & dosagem , Plasmídeos/administração & dosagem , Tomografia por Emissão de Pósitrons , Compostos de Amônio Quaternário/administração & dosagem , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteínas Supressoras de Tumor/administração & dosagem , Proteínas Supressoras de Tumor/metabolismoRESUMO
Unrelated cord blood transplant (CBT) is an alternative treatment option for patients who lack a matched donor. However, the optimal type and intensity of the preparative regimen remains unclear. We evaluated the toxicity and outcomes of a conditioning regimen consisting of melphalan 140 mg/m(2) (day - 8), thiotepa 10 mg/kg (day - 7), fludarabine 160 mg/m(2) over 4 days (days - 6 to - 3) and rabbit antithymocyte globulin (ATG) 1.25 mg/kg (day - 4) and 1.75 mg/kg (day - 3) (FMT). Forty-seven patients with advanced hematologic malignancies with a median age of 23 years (30 adults and 17 children) were treated. Sixty percent of patients were in remission at transplant. Ninety-one percent of the patients engrafted neutrophils after a median of 22 days, and all but one of the patients achieving donor engraftment had hematopoietic recovery with 100% cord blood-derived cells. Grade 3 gastrointestinal toxicity was the major non-hematopoietic toxicity, occurring in 32% of patients. Cumulative incidence of day-100 grade II-IV acute graft-versus-host disease (aGVHD) and chronic graft-versus-host disease (cGVHD) was 53% and 34%, respectively, and non-relapse mortality at day 100 and 2 years was 11% and 40%. Two-year disease-free and overall survival rates were 31% and 44%, respectively. These results suggest that FMT is a feasible conditioning regimen for patients undergoing CBT.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Transplante de Células-Tronco de Sangue do Cordão Umbilical/métodos , Neoplasias Hematológicas/terapia , Condicionamento Pré-Transplante/métodos , Adolescente , Adulto , Soro Antilinfocitário/administração & dosagem , Estudos de Viabilidade , Sobrevivência de Enxerto , Doença Enxerto-Hospedeiro , Humanos , Melfalan/administração & dosagem , Taxa de Sobrevida , Tiotepa/administração & dosagem , Vidarabina/administração & dosagem , Vidarabina/análogos & derivados , Adulto JovemRESUMO
PURPOSE: Adoptive cell therapy (ACT) using autologous tumor-infiltrating lymphocytes (TIL) is a promising treatment for metastatic melanoma unresponsive to conventional therapies. We report here on the results of an ongoing phase II clinical trial testing the efficacy of ACT using TIL in patients with metastatic melanoma and the association of specific patient clinical characteristics and the phenotypic attributes of the infused TIL with clinical response. EXPERIMENTAL DESIGN: Altogether, 31 transiently lymphodepleted patients were treated with their expanded TIL, followed by two cycles of high-dose interleukin (IL)-2 therapy. The effects of patient clinical features and the phenotypes of the T cells infused on the clinical response were determined. RESULTS: Overall, 15 of 31 (48.4%) patients had an objective clinical response using immune-related response criteria (irRC) with 2 patients (6.5%) having a complete response. Progression-free survival of more than 12 months was observed for 9 of 15 (60%) of the responding patients. Factors significantly associated with the objective tumor regression included a higher number of TIL infused, a higher proportion of CD8(+) T cells in the infusion product, a more differentiated effector phenotype of the CD8(+) population, and a higher frequency of CD8(+) T cells coexpressing the negative costimulation molecule "B- and T-lymphocyte attenuator" (BTLA). No significant difference in the telomere lengths of TIL between responders and nonresponders was identified. CONCLUSION: These results indicate that the immunotherapy with expanded autologous TIL is capable of achieving durable clinical responses in patients with metastatic melanoma and that CD8(+) T cells in the infused TIL, particularly differentiated effectors cells and cells expressing BTLA, are associated with tumor regression.