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Luis Ignacio Sánchez-Abarca,
Isabel Alvarez-Laderas,
María Díez Campelo,
Teresa Caballero-Velázquez,
Carmen Herrero,
Sandra Muntión,
Cristina Calderón,
Estefanía García-Guerrero,
Fermín Sánchez-Guijo, Consuelo Del Cañizo,
Jesús San Miguel,
José Antonio Pérez-Simón
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ABSTRACT: BACKGROUND AIMS: Mesenchymal stromal cells (MSCs) are multipotent stem cells with immunosuppressive properties. Nevertheless, it has been previously reported that MSCs might also trigger the immune response. We studied whether MSCs may act as carriers, capturing antigens that can be endocytosed by antigen-presenting cells later on. METHODS: We measured the cellular uptake of mannose receptor-mediated fluid phase macropinocytosis, assessed as cellular uptake of fluorescein isothiocyanate-dextran, and PKH-67-labeled cell lysates as a surrogate marker for antigen capture among dendritic cells (DCs, positive control), T lymphocytes (negative control) and MSCs. RESULTS: All experiments confirmed that MCSs displayed pinocytic and endocytic capacities, which were lower than those observed for DCs but significantly higher than those observed for T cells. We also demonstrated that MSCs release previously endocytosed antigens, which subsequently can be captured by DCs. CONCLUSIONS: MSCs have the ability to capture and release antigens.
Cytotherapy 03/2013; · 3.63 Impact Factor
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Carlos Santamaría,
Fernando Ramos,
Noemi Puig,
Eva Barragán,
Raquel de Paz,
Carme Pedro,
Andrés Insunza,
Mar Tormo, Consuelo Del Cañizo,
María Diez-Campelo,
Blanca Xicoy,
Eduardo Salido,
Javier Sánchez Del Real,
Montserrat Hernández,
Carmen Chillón,
Guillermo F Sanz,
Ramón García-Sanz,
Jesús F San Miguel,
Marcos González
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ABSTRACT: Several studies have evaluated the prognostic value of the individual expression of certain genes in patients with myelodysplastic syndromes (MDS). However, none of them includes their simultaneous analysis by quantitative polymerase chain reaction (PCR). We evaluated relative expression levels of 14 molecular markers in 193 peripheral blood samples from untreated MDS patients using real-time PCR. Detectable WT1 expression levels, low TET2, and low IER3 gene expression were the only markers showing in univariate analysis a poor prognostic value for all treatment-free (TFS), progression-free (PFS), and overall survival (OS). In multivariate analysis, molecular parameters associated with a shorter TFS were: WT1 detection (p = 0.014), low TET2 (p = 0.002), and low IER3 expression (p = 0.025). WT1 detection (p = 0.006) and low TET2 (p = 0.006) expression were associated with a shorter PFS when multivariate analysis was carried out by including only molecular markers. Molecular values with an independent value in OS were: WT1 detection (p = 0.003), high EVI1 expression (p = 0.001), and undetectatable p15-CDKN2B (p = 0.037). WT1 expressers were associated with adverse clinical-biological features, high IPSS and WPSS scoring, and unfavorable molecular expression profile. In summary, detectable WT1 expression levels, and low TET2 and low IER3 expression in peripheral blood showed a strong association with adverse prognosis in MDS patients at diagnosis. However, WT1 was the only molecular marker displaying an independent prognostic value in both OS and TFS.
Annals of Hematology 08/2012; · 2.62 Impact Factor
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ABSTRACT: The onset of myelodysplastic syndromes (MDS) is usually around the age of 70. Despite this, most clinical trials are restricted to younger subjects. Thus, the management of elderly patients with MDS is not always optimal. Physiologically, elderly patients show characteristics that differ from those of younger patients and that condition their pharmacological treatment. In this regard, the comprehensive geriatric assessment (CGA) becomes particularly important. This document gathers conclusions from the 1(st) Meeting of Members of the Sociedad Española de Medicina Geriátrica and the Sociedad Española de Hematología y Hemoterapia, with the objective of proposing the establishment of CGA instruments to assist in the decision-making process of elderly patients with MDS. The results of this consensus document will focus on the diagnosis, prognosis, treatment and management of adverse events in this age group.
Medicina Clínica 02/2012; 138(3):119.e1-9. · 1.38 Impact Factor
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Iris Cordoba,
José R González-Porras,
Benet Nomdedeu,
Elisa Luño,
Raquel de Paz,
Esperanza Such,
Mar Tormo,
Teresa Vallespi,
Rosa Collado,
Blanca Xicoy,
Rafael Andreu,
Juan A Muñoz,
Francesc Solé,
Jose Cervera, Consuelo del Cañizo
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ABSTRACT: Abnormalities involving chromosome 7 are frequent in myelodysplastic syndrome (MDS) and suggest a poor prognosis.
The authors examined the hypothesis that the clinical features and survival associated with isolated deletion (del) of part of the long arm of chromosome 7 (7q) in MDS are different from those associated with isolated monosomy 7 (complete loss of chromosome 7). In total, 133 patients with a diagnosis of de novo MDS (according to the World Health Organization [WHO] classification) and chromosome 7 abnormalities in the Spanish MDS Registry were evaluated retrospectively. Four karyotypic groups were identified: isolated del(7q) (n = 29), isolated monosomy 7 (n = 27), del(7q) with additional abnormalities (n = 24), and monosomy 7 with additional abnormalities (n = 53).
Isolated del(7q) was more frequent in patients with less advanced MDS according to the WHO classification or the International Prognostic Scoring System. In addition, isolated del(7q) was associated with fewer blasts in bone marrow than other cytogenetics groups. Survival was significantly superior in patients with isolated del(7) than in those with isolated monosomy 7, del(7q) with additional abnormalities, or monosomy 7 with additional abnormalities. On multivariate analysis, age, the percentage of blasts in bone marrow, and other chromosome 7 abnormalities apart from isolated del(7q) were identified as independent risk factors for survival.
The current results demonstrated that patients who had MDS with isolated del(7q) had some distinct clinical-pathologic characteristics as well as better survival than patients who had MDS with isolated monosomy 7.
Cancer 06/2011; 118(1):127-33. · 4.77 Impact Factor
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ABSTRACT: The clone size has been postulated as a prognostic factor in myelodysplastic syndromes (MDS), though it has not been studied systematically. We tested its impact (<100% vs. 100%) in a population of 216 MDS with chromosome 7 abnormalities (-7/7q-) (n=84), trisomy 8 (n=99), 20q deletion (n=28) and loss of Y chromosome (n=26). Focusing on the survival the bad prognosis of -7/7q- was independent of the clone size (9.3 vs. 5.0 months, P=0.188, not significant) but trisomy 8 cases with 100% aberrant metaphases did reveal a worse prognosis (13.9 vs. 5.9 months, P=0.003).
Leukemia research 01/2011; 35(6):834-6. · 2.36 Impact Factor
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Fernando Silva,
José A Pérez-Simón,
Teresa Caballero Velazquez,
Cristina Encinas,
Fermín M Sánchez-Guijo,
María Díez-Campelo,
Enrique Colado,
Jesús Martín,
Fernanda Villanueva-Gomez,
Lourdes Vazquez, Consuelo Del Cañizo,
Dolores Caballero,
Jesús San Miguel
American Journal of Hematology 12/2009; 85(4):290-3. · 4.67 Impact Factor
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Belén Blanco,
José A Pérez-Simón,
Luis I Sánchez-Abarca,
Teresa Caballero-Velazquez,
Silvia Gutierrez-Cossío,
Pilar Hernández-Campo,
María Díez-Campelo,
Carmen Herrero-Sanchez,
Concepción Rodriguez-Serrano,
Carlos Santamaría,
Fermín M Sánchez-Guijo, Consuelo Del Cañizo,
Jesús F San Miguel
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ABSTRACT: In vitro depletion of alloreactive T cells using the proteasome inhibitor bortezomib is a promising approach to prevent graft-versus-host disease after allogeneic stem cell transplantation. We have previously described the ability of bortezomib to selectively eliminate alloreactive T cells in a mixed leukocyte culture, preserving non-activated T cells. Due to the role of regulatory T cells in the control of graft versus host disease, in the current manuscript we have analyzed the effect of bortezomib in regulatory T cells.
Conventional or regulatory CD4(+) T cells were isolated with immunomagnetic microbeads based on the expression of CD4 and CD25. The effect of bortezomib on T-cell viability was analyzed by flow cytometry using 7-amino-actinomycin D staining. To investigate the possibility of obtaining an enriched regulatory T-cell population in vitro with the use of bortezomib, CD4(+) T cells were cultured during four weeks in the presence of anti-CD3 and anti-CD28 antibodies, IL-2 and bortezomib. The phenotype of these long-term cultured cells was studied, analyzing the expression of CD25, CD127 and FOXP3 by flow cytometry, and mRNA levels were determined by RT-PCR. Their suppressive capacity was assessed in co-culture experiments, analyzing proliferation and IFN-gamma and CD40L expression of stimulated responder T cells by flow cytometry.
We observed that naturally occurring CD4(+)CD25(+) regulatory T cells are resistant to the pro-apoptotic effect of bortezomib. Furthermore, we found that long-term culture of CD4(+) T cells in the presence of bortezomib promotes the emergence of a regulatory T-cell population that significantly inhibits proliferation, IFN-gamma production and CD40L expression among stimulated effector T cells.
These results reinforce the proposal of using bortezomib in the prevention of graft versus host disease and, moreover, in the generation of regulatory T-cell populations, that could be used in the treatment of multiple T-cell mediated diseases.
Haematologica 07/2009; 94(7):975-83. · 6.42 Impact Factor
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Maitane Pérez-Ilzarbe,
María Díez-Campelo,
Pablo Aranda,
Soraya Tabera,
Tania Lopez, Consuelo del Cañizo,
Juana Merino,
Cristina Moreno,
Enrique J Andreu,
Felipe Prósper,
José Antonio Pérez-Simón
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ABSTRACT: Mesenchymal stem cells (MSCs) are multipotent stem cells. Based on their properties, several clinical trials have been designed to explore their potential therapeutic effect. Fetal calf serum (FCS, commonly used for in vitro expansion) is an undesirable source of xenogeneic antigens and bears the risk of transmitting contaminations. As an alternative for FCS, platelet lysate (PL) and both autologous and allogeneic human serum have been proposed. The aim of this study is to compare the culture of bone marrow (BM)-derived MSCs in the presence of different serum supplements to determine the effect on cell growth, differentiation potential, and immunologic function.
MSCs from BM of healthy volunteer donors were grown in the presence of 10% FCS supplemented with 1 ng/mL basic fibroblast growth factor (bFGF), 10% human serum supplemented with 1 ng/mL bFGF, 5% PL, and PL 5% supplemented with 1 ng/mL bFGF (PL plus bFGF).
MSCs that expanded in either medium showed a comparable morphology, phenotype, and proliferative and differentiation capacity. While the presence of MSCs in vitro significantly decreased CD3/CD28-mediated T-cell activation, this effect was significantly higher in MSCs cultured with human serum. Production of interferon-gamma was inhibited by cocultured media with MSCs while MSCs also induced a significant inhibition of cell cycle in T cells.
In conclusion, PL or autologous serum could offer an alternative to the use of FCS in MSC expansion for clinical use maintaining the same growing potential, phenotype, immunomodulatory properties, and differentiation potential.
Transfusion 06/2009; 49(9):1901-10. · 3.22 Impact Factor
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José A Pérez-Simón,
Cristina Encinas,
Fernando Silva,
Maria José Arcos,
María Díez-Campelo,
Fermín M Sánchez-Guijo,
Enrique Colado,
Jesús Martín,
Lourdes Vazquez, Consuelo del Cañizo,
Dolores Caballero,
Jesús San Miguel
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ABSTRACT: Several grading systems have been developed in the bone marrow transplantation setting in attempts to predict survival in patients with chronic graft-versus-host disease (cGVHD). In this study, we evaluated the prognostic value of the National Institutes of Health (NIH) scoring system and investigated for any additional prognostic factors in a series of 171 patients undergoing peripheral blood stem cell transplantation (PBSCT) from matched related donors. The cumulative incidence of cGVHD was 70%; cumulative incidences of mild, moderate, and severe cGVHD were 29%, 42% and 28%, respectively. Overall, 68% of patients were free from immunosuppression 5 years after transplantation. Absence of previous acute GVHD (aGVHD; hazard ratio [HR] = 2; P = .004) and mild cGVHD (HR = 4.2; P = .007) increased the probability of being off immunosuppressive treatment by the last follow-up. Overall survival (OS) at 5 years was 52%. Severe cGVHD, according to the NIH scoring system (HR = 13.27; P = .001) adversely influenced outcome, whereas de novo onset (HR = 0.094; P = .003) had a more favorable impact on survival. The combination of both variables allowed us to identify 4 different subgroups of patients with OS of 82%, 70%, 50%, and 25%. Our findings indicate that the NIH scoring system has some prognostic value in patients undergoing PBSCT and, together with the type of onset, must be considered to predict the possible outcome of patients who develop cGVHD.
Biology of blood and marrow transplantation: journal of the American Society for Blood and Marrow Transplantation 11/2008; 14(10):1163-71. · 3.15 Impact Factor
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ABSTRACT: Since the feasibility of stem cell therapy has been recognized, enthusiasm for this therapy has grown exponentially. Nevertheless, as professionals we must realize that this enthusiasm should relate not only to our scientific interest but also to the care of our patients. Within the next decade, patients' demand for the latest therapies is likely to rise because of changes in health care systems that will broaden availability. Stem cell therapy is likely to be among these in-demand treatments, and we must be prepared for this change. In this Review we discuss the basic principles of how to launch a clinical program for stem cell therapy for cardiovascular repair. First, we look at the composition of the program team. Second, we describe the different types of stem cells available in clinical practice. Third, we present in depth the two most widely applicable delivery approaches. Finally, we discuss selection of patients and approaches and clinical and imaging methods by which to evaluate the safety and efficacy of this therapy.
Nature Clinical Practice Cardiovascular Medicine 03/2007; 4 Suppl 1:S123-9. · 7.04 Impact Factor
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ABSTRACT: Four-color multiparameter immunophenotyping has recently proven to be an attractive technique for evaluating the plasma cell (PC) compartment since it allows discrimination between myelomatous and normal PC. This study was designed to investigate: i) whether peripheral blood is less contaminated than bone marrow as a source for an autologous transplant; ii) the effect of growth factors on mobilizing myelomatous PC into peripheral blood; iii) the degree of contamination by myelomatous PC in apheresis samples; and iv) whether the number of PC increases during the last days of apheresis.
Using 4-color antigen staining we investigated the composition of the PC compartment in 90 apheresis products from 40 patients with MM; in 17 cases bone marrow and peripheral blood samples were also simultaneously evaluated.
(i) All pre-mobilization bone marrow samples analyzed were always contaminated with myelomatous PC whereas only 41% of the post-mobilization peripheral blood samples were contaminated. Moreover, the use of peripheral blood would lead to a reduction of >5x10(5) infused myelomatous PC; (ii) mobilization with cytokines increased the number of circulating PC, generally because of an expansion of the normal PC population; (iii) forty-eight percent of all peripheral blood stem cell harvests were contaminated with myelomatous PC, although normal PC usually represented the predominant population; (iv) no significant changes were observed in the amount of contaminating myelomatous PC during the first three days of apheresis.
Multiparameter immunophenotyping is a useful approach for investigating the PC compartment in apheresis products.
Haematologica 10/2003; 88(9):1013-21. · 6.42 Impact Factor
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Miguel A Díaz,
Julián Sevilla,
Javier de la Rubia,
Amparo Verdeguer,
Ildefonso Espigado,
Marta G Vicent,
Maria J Pascual,
Concha Zamora,
Rosario Arrieta,
David Serrano, Consuelo del Cañizo,
Cristina Arbona,
Felipe de Arriba,
Joan Bargay,
Salut Brunet,
Miguel A Sanz
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ABSTRACT: Although several studies have reported on the use of children as donors for peripheral blood progenitor cells (PBPC), no specific characteristics have been identified as predictors of PBPC collection in this population. In this study we analyzed predictive factors for PBPC collection in pediatric donors.
We retrospectively analyzed factors predicting the yield for a target CD34+ cell dose of > or =4x10(6)/Kg donor or recipient body weight, in 105 aphereses from 76 healthy pediatric donors (36 boys and 40 girls) included in the Spanish National Donor Registry. Mobilization consisted of granulocyte colony-stimulating factor (G-CSF) in single doses of 10 microg/kg per day subcutaneously for 4 or 5 days. Apheresis started after the fourth dose of G-CSF.
Median age and body weight were 10 years (range 1-18) and 42 kg (range 9-89), respectively. The median number of CD34+ cells/kg recipient body weight was 4.22 (range 0.1-32). On multivariate analysis variables that had a significant negative impact on the CD34+ cell yield, considering the recipient's body weight were the total blood volume processed (regression coefficient (RC): 0.41, 95% CI: 0.21-0.81; p=0.01) and day of apheresis other than first (RC: 0.16, 95% CI: 0.07-0.34; p<0.0001). When considering donor's body weight the variables that positively influenced collection were younger age (RC: 6.79, 95% CI: 1.57-29.25; p<0.01) and large volume leukapheresis (RC: 3.33, 95% CI: 1.13-9.77; p<0.02).
Our data suggest that pediatric donors mobilized by G-CSF may donate sufficient numbers of CD34+ cells for allogeneic transplantation. The variables that influenced the yield were the donor's age, blood volume processed and the first day of the apheresis.
Haematologica 08/2003; 88(8):919-22. · 6.42 Impact Factor
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Javier De la Rubia,
Cristina Arbona, Consuelo Del Cañizo,
Rosario Arrieta,
Felipe De Arriba,
María J Pascual,
Isabel Sanjuan,
Miguel A Díaz,
Salut Brunet,
Adrián Alegre, [......],
Javier De la Serna,
David Serrano,
Joan Bargay,
José Petit,
Dobleta Martínez,
Amparo Verdeguer,
José M Ribera,
Carmen Martínez,
Luis Benlloch,
Miguel A Sanz
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ABSTRACT: We have retrospectively evaluated the results of two cycles of mobilization and collection of peripheral blood progenitor cells (PBPC) from 46 healthy donors included in the Spanish National Donor Registry. Mobilization involved the administration of granulocyte colony-stimulating factor (G-CSF) at a median dose of 10 microg/kg per day, and apheresis was begun after the fourth dose of G-CSF in both cycles. The median interval between both mobilizations was 187 days (range, 7-1428 days). The incidence and types of side-effects were similar after both donations, with 25 and 26 donors developing some toxicity after the first and second donations, respectively. The median number of CD34(+) cells collected was higher after the first mobilization than after the second (5.15 versus 3.16 x 10(6)/kg, respectively; p = 0.05), and 29 donors yielded fewer CD34(+) cells after the second mobilization (p = 0.018). A lower proportion of donors yielded CD34(+) cell counts >4 x 10(6)/kg after the second cycle than after the first (52% versus 76%, respectively; p = 0.057). Our study shows that second rounds of PBPC collection from normal donors are well tolerated but are associated with a significantly reduced number of CD34(+) cells collected when the same mobilization scheme is used.
Journal of Hematotherapy & Stem Cell Research 08/2002; 11(4):705-9.
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Javier de la Rubia,
Cristina Arbona,
Felipe de Arriba, Consuelo del Cañizo,
Salut Brunet,
Concha Zamora,
Miguel A Díaz,
Joan Bargay,
José Petit,
Javier de la Serna, [......],
María J Pascual,
David Serrano,
Isabel Sanjuan,
Ildefonso Espigado,
Adrián Alegre,
Dobleta Martínez,
Amparo Verdeguer,
Carmen Martínez,
Luis Benlloch,
Miguel A Sanz
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ABSTRACT: Predictive factors of the response to rHuG-CSF in normal donors have not been extensively studied.
We analyzed factors influencing CD34+ cell yield in the 1st day of collection in 261 healthy donors from the Spanish National Donor Registry. The median age was 38 years (range, 2-72). The median dose of rHuG-CSF was 10 microg per kg per day (range, 5-20) over 4 days. In 103 donors (40%), <4 x 10(6) per kg CD34+ cells were collected. The variables that were analyzed included age, sex, weight, basal complete blood cell count, dose, type of rHuGCSF and schedule of administration, and maximum WBC count before apheresis.
By univariate analysis, the maximum WBC count (<50 vs. >or=50 x 10(9)/L, p = 0.004), advanced age (p = 0.008), and number of daily rHuG-CSF doses (one vs. two; p = 0.01) correlated with the number of CD34+ cells collected. By multivariate analysis, donors age (<38 vs. >or=38 years; p = 0.014) and a single daily dose of rHuG-CSF (p = 0.005) were the two variables that significantly predicted a low CD34+ cell yield.
Donors' age, with a threshold of 38 years or more, and the rHuG-CSF schedule are the factors that significantly affected CD34+ cell mobilization and collection in healthy donors.
Transfusion 01/2002; 42(1):4-9. · 3.22 Impact Factor
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ABSTRACT: In acute non-lymphoblastic leukemia (ANLL) progenitor cells frequently display a certain degree of autonomous growth. The aim of the present work was to analyze the autonomous proliterative capacity of leukemic progenitors in both de novo and secondary to myeloproliferative disorders (MPD) and myelodysplastic syndromes (MDS), acute myeloid leukemias and to correlate with clinical and biological characteristics of the disease. Clonogenic assays with and without leukocyte conditioned medium with PHA (LCM-PHA) were performed and the autonomous proliferation index (API) calculated in a series of 50 patients (34 de novo ANLL, eight secondary to MPD and eight secondary to MDS). Patients were divided into two groups according to their API, low (⩽0.4) or high (>0.4). Autonomous growth was observed in 84% of cases studied (82% in de novo ANLL, 75% secondary to MDS and 100% secondary to MPD). The group with the highest API (29 patients) had increased levels of hemoglobin (P = 0.006) and platelets (P = 0.01). A high API was also associated with an immature phenotype of blast cells (P = 0.02). Upon analyzing the de novo ANLL separately we observed that a high API correlated with high Hb values (P = 0.02), a lower rate of complete remission (42% vs 61%) and a lower survival rate (medium of 3 vs 10 months). These findings suggest that the capacity for autonomous proliferation can condition the clinical and biological profile of the disease.
Leukemia Research.
