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1.
Eur Arch Otorhinolaryngol ; 280(10): 4519-4530, 2023 Oct.
Article in English | MEDLINE | ID: mdl-37254001

ABSTRACT

PURPOSE: Whilst immunotherapy is an appealing option as it could reduce the burden of recurrent pediatric respiratory tract infections (RTI), there is limited evidence on its effectiveness and more research was requested in order to better understand this therapeutic modality. METHODS: We performed a prospective cohort study involving 57 subjects to assess the safety and effectiveness a 3-month regimen of either typified or patient-specific bacterial lysates could have in reducing the number of RTIs in children aged 0 to 11 years with histories of recurrent episodes. RESULTS: After a 6-month follow-up, the number of RTIs and school absenteeism dropped sharply and significantly, from an adjusted mean (standard error) of 0.6 (0.04) episodes/month to 0.1 (0.03) episodes/month (74.7% reduction, P < 0.001), and from an adjusted mean score of 4.6 (1.06) points to 0.0 (0.01) points over 10 (99.5% reduction, P < 0.001), respectively. There was also a significant decrease in the severity of symptoms. No adverse reactions were observed. CONCLUSION: The use of the study product is associated with a decreased risk of recurrent RTIs in children, with a very favorable safety profile that warrants further investigation in randomized clinical trials.


Subject(s)
Respiratory Tract Infections , Child , Humans , Prospective Studies , Respiratory Tract Infections/prevention & control , Immunotherapy , Absenteeism , Bacteria
2.
Dis Model Mech ; 15(3)2022 03 01.
Article in English | MEDLINE | ID: mdl-35191981

ABSTRACT

Pearson syndrome is a rare multisystem disease caused by single large-scale mitochondrial DNA deletions (SLSMDs). The syndrome presents early in infancy and is mainly characterised by refractory sideroblastic anaemia. Prognosis is poor and treatment is supportive, thus the development of new models for the study of Pearson syndrome and new therapy strategies is essential. In this work, we report three different cell models carrying an SLMSD: fibroblasts, transmitochondrial cybrids and induced pluripotent stem cells (iPSCs). All studied models exhibited an aberrant mitochondrial ultrastructure and defective oxidative phosphorylation system function, showing a decrease in different parameters, such as mitochondrial ATP, respiratory complex IV activity and quantity or oxygen consumption. Despite this, iPSCs harbouring 'common deletion' were able to differentiate into three germ layers. Additionally, cybrid clones only showed mitochondrial dysfunction when heteroplasmy level reached 70%. Some differences observed among models may depend on their metabolic profile; therefore, we consider that these three models are useful for the in vitro study of Pearson syndrome, as well as for testing new specific therapies. This article has an associated First Person interview with the first author of the paper.


Subject(s)
Lipid Metabolism, Inborn Errors , Mitochondrial Diseases , Congenital Bone Marrow Failure Syndromes/genetics , DNA, Mitochondrial/genetics , Humans , Lipid Metabolism, Inborn Errors/genetics , Mitochondrial Diseases/genetics , Muscular Diseases
3.
J Clin Pathol ; 74(3): 198-201, 2021 Mar.
Article in English | MEDLINE | ID: mdl-32796052

ABSTRACT

Screening of haemoglobinopathies is indicated for the detection of sickle cell anaemia; thus, neonates can benefit from early and adequate treatment that prevents neurological damage, reduces morbidity and mortality associated with the disease. These types of programmes sometimes lead to unexpected findings. We present a new haemoglobin (Hb) variant (Hb Miguel Servet) detected by newborn screening. During neonatal screening of haemoglobinopathies by cation-exchange high-performance liquid chromatography (CE-HPLC) newborn, an Hb variant was detected. An analysis at 8 months of age using capillary zone electrophoresis (CZE) confirmed the presence of this new Hb. The molecular characterisation was performed by automatic sequencing of the α and ß globin genes in an ABI PRISM 3100 Genetic Analyzer. Hb analysis by CE-HPLC ß-thalassaemia short programmedid not indicate the presence of abnormal Hbs. By CZE showed a peak in the zone 12 zone comprising 3.3% of the total Hb. A new analysis by CE-HPLC on a Tosoh G8-2 (Horiba) shown a peak, in the region of HbA1b, did not interfere with the quantification of HbA1c. Sequencing of the ß gene revealed the substitution of a guanine for a thymine (GGT >TGT) in codon 69 of the second exon, resulting in substitution of cysteine for the amino acid glutamine (HBB:c.208G>T). Hb Miguel Servet is a ß-chain globin variant detected by CE-HPLC newborn (BioRad), by CZE and by CE-HPLC-CE Tosoh G8-2 (Horiba), but no by CE-HPLC-CE ß-thalassaemia short programme (BioRad). In fact, for all the techniques that are visualised, what would be detected would be the glutathione variant of Hb (Miguel Servet).


Subject(s)
Anemia, Sickle Cell/diagnosis , Hemoglobinopathies/diagnosis , beta-Thalassemia/diagnosis , Anemia, Sickle Cell/genetics , Anemia, Sickle Cell/pathology , Chromatography, High Pressure Liquid , Electrophoresis, Capillary , Hemoglobinopathies/genetics , Hemoglobinopathies/pathology , Humans , Male , beta-Globins/genetics , beta-Thalassemia/genetics , beta-Thalassemia/pathology
5.
Clin Nucl Med ; 45(2): e122-e124, 2020 Feb.
Article in English | MEDLINE | ID: mdl-31714280

ABSTRACT

Immunoglobulin G4-related disease (IgG4-RD) is a systemic inflammatory condition that can manifest in atypical locations. We present the case of a middle-aged woman presenting with isolated IgG4-RD of left maxilla and nasal septum, confirmed with F-FDG PET/CT. She achieved complete metabolic response with multiple pharmacologic treatment, but 1 year later, she experienced local relapse, and FDG PET/CT found new additional lesions in humeral vasculature and lungs. This case illustrates the usefulness of FDG PET/CT for initial evaluation and monitoring of IgG4-RD with unusual nasomaxillary involvement.


Subject(s)
Fluorodeoxyglucose F18 , Immunoglobulin G4-Related Disease/diagnostic imaging , Maxilla/diagnostic imaging , Nasal Septum/diagnostic imaging , Positron Emission Tomography Computed Tomography , Female , Follow-Up Studies , Humans , Middle Aged , Recurrence
6.
Orphanet J Rare Dis ; 14(1): 82, 2019 04 17.
Article in English | MEDLINE | ID: mdl-30995915

ABSTRACT

BACKGROUND: Telomeres are nucleoprotein structures present at the terminal region of the chromosomes. Mutations in genes coding for proteins involved in telomere maintenance are causative of a number of disorders known as telomeropathies. The genetic origin of these diseases is heterogeneous and has not been determined for a significant proportion of patients. METHODS: This article describes the genetic characterization of a cohort of patients. Telomere length was determined by Southern blot and quantitative PCR. Nucleotide variants were analyzed either by high-resolution melting analysis and Sanger sequencing of selected exons or by massive sequencing of a panel of genes. RESULTS: Forty-seven patients with telomere length below the 10% of normal population, affected with three telomeropathies: dyskeratosis congenita (4), aplastic anemia (22) or pulmonary fibrosis (21) were analyzed. Eighteen of these patients presented known pathogenic or novel possibly pathogenic variants in the telomere-related genes TERT, TERC, RTEL1, CTC1 and ACD. In addition, the analyses of a panel of 188 genes related to haematological disorders indicated that a relevant proportion of the patients (up to 35%) presented rare variants in genes related to DNA repair or in genes coding for proteins involved in the resolution of complex DNA structures, that participate in telomere replication. Mutations in some of these genes are causative of several syndromes previously associated to telomere shortening. CONCLUSION: Novel variants in telomere, DNA repair and replication genes are described that might indicate the contribution of variants in these genes to the development of telomeropathies. Patients carrying variants in telomere-related genes presented worse evolution after diagnosis than the rest of patients analyzed.


Subject(s)
Anemia, Aplastic/genetics , DNA Repair/genetics , Dyskeratosis Congenita/genetics , Pulmonary Fibrosis/genetics , Telomere Shortening/genetics , Telomere/genetics , Adolescent , Adult , Child , Child, Preschool , Exons/genetics , Female , Humans , Infant , Male , Pedigree , RNA/genetics , Telomerase/genetics , Young Adult
7.
Front Immunol ; 8: 802, 2017.
Article in English | MEDLINE | ID: mdl-28747912

ABSTRACT

GATA binding protein 2 (GATA2) deficiency is a rare disorder of hematopoiesis, lymphatics, and immunity caused by spontaneous or autosomal dominant mutations in the GATA2 gene. Clinical manifestations range from neutropenia, lymphedema, deafness, to severe viral and mycobacterial infections, bone marrow failure, and acute myeloid leukemia. Patients also present with monocytopenia, dendritic cell, B- and natural killer (NK)-cell deficiency. We studied the T-cell and NK-cell compartments of four GATA2-deficient patients to assess if changes in these lymphocyte populations could be correlated with clinical phenotype. Patients with more severe clinical complications demonstrated a senescent T-cell phenotype whereas patients with lower clinical score had undetectable changes relative to controls. In contrast, patients' NK-cells demonstrated an immature/activated phenotype that did not correlate with clinical score, suggesting an intrinsic NK-cell defect. These studies will help us to determine the contribution of T- and NK-cell dysregulation to the clinical phenotype of GATA2 patients, and may help to establish the most accurate therapeutic options for these patients. Asymptomatic patients may be taken into consideration for hematopoietic stem cell transplantation when dysregulation of T-cell and NK-cell compartment is present.

9.
Nutr Hosp ; 33(3): 260, 2016 Jun 30.
Article in Spanish | MEDLINE | ID: mdl-27513487

ABSTRACT

Introducción: los supervivientes de leucemia aguda (LA) infantil presentan un riesgo incrementado de alteraciones metabólicas y cardiovasculares que aumentan su morbimortalidad a largo plazo.Objetivo: estimar la prevalencia de obesidad, resistencia a la insulina, dislipemia e hipertensión arterial como factores de riesgo cardiometabólico (FRCM) en un grupo de supervivientes de LA infantil, y analizar las posibles causas asociadas a su desarrollo.Material y métodos: estudio observacional retrospectivo en 47 supervivientes de LA tratados en un periodo de 4 años, que recibieron seguimiento durante 10 años.Resultados: el 40% de los participantes presentaron al menos un FRCM durante el seguimiento, siendo la dislipemia (aumento LDL) el más frecuente (38,3%), seguido de obesidad/sobrepeso (31,9%) y HTA sistólica (23,4%). El sexo femenino se estableció como factor de riesgo parael desarrollo de todos ellos (RR 1,6; RR 3,16; RR 1,69; p < 0,05). Ningún superviviente desarrolló diabetes mellitus, pero sí resistencia a la insulina el 19,4%. Los pacientes con leucemias de peor pronóstico presentaron mayor riesgo de desarrollar obesidad, resistencia a la insulina y aumento de LDL (RR 3,56; RR 4,08; RR 2,53; p < 0,05). Los pacientes tratados con trasplante de progenitores hematopoyéticos presentaron mayor riesgo de obesidad, aumento de LDL e HTA sistólica (RR 2,86; RR 2,39; RR 3,12; p<0,05). La radioterapia se asoció de igual modo con un incremento de resistencia a la insulina e hipertensión arterial sistólica (RR 2,47; RR 2,53; p < 0,05).Conclusiones: existe un aumento en la prevalencia de obesidad/sobrepeso, dislipemia, resistencia a la insulina y alteración de la tensión arterial sistólica en supervivientes de leucemia aguda infantil a lo largo del tiempo, especialmente en aquellos con enfermedades y tratamientos más agresivos.


Subject(s)
Cardiovascular Diseases/etiology , Leukemia/complications , Metabolic Diseases/etiology , Survivors , Adolescent , Cardiovascular Diseases/epidemiology , Child , Child, Preschool , Female , Humans , Infant , Insulin Resistance , Leukemia/epidemiology , Lost to Follow-Up , Male , Metabolic Diseases/epidemiology , Obesity/epidemiology , Obesity/etiology , Overweight/epidemiology , Overweight/etiology , Prevalence , Risk Factors , Young Adult
10.
Nutr. hosp ; 33(3): 549-555, mayo-jun. 2016. tab, graf
Article in Spanish | IBECS | ID: ibc-154470

ABSTRACT

Introducción: los supervivientes de leucemia aguda (LA) infantil presentan un riesgo incrementado de alteraciones metabólicas y cardiovasculares que aumentan su morbimortalidad a largo plazo. Objetivo: estimar la prevalencia de obesidad, resistencia a la insulina, dislipemia e hipertensión arterial como factores de riesgo cardiometabólico (FRCM) en un grupo de supervivientes de LA infantil, y analizar las posibles causas asociadas a su desarrollo. Material y métodos: estudio observacional retrospectivo en 47 supervivientes de LA tratados en un periodo de 4 años, que recibieron seguimiento durante 10 años. Resultados: el 40% de los participantes presentaron al menos un FRCM durante el seguimiento, siendo la dislipemia (aumento LDL) el más frecuente (38,3%), seguido de obesidad/sobrepeso (31,9%) y HTA sistólica (23,4%). El sexo femenino se estableció como factor de riesgo para el desarrollo de todos ellos (RR 1,6; RR 3,16; RR 1,69; p < 0,05). Ningún superviviente desarrolló diabetes mellitus, pero sí resistencia a la insulina el 19,4%. Los pacientes con leucemias de peor pronóstico presentaron mayor riesgo de desarrollar obesidad, resistencia a la insulina y aumento de LDL (RR 3,56; RR 4,08; RR 2,53; p < 0,05). Los pacientes tratados con trasplante de progenitores hematopoyéticos presentaron mayor riesgo de obesidad, aumento de LDL e HTA sistólica (RR 2,86; RR 2,39; RR 3,12; p< 0,05). Conclusiones: existe un aumento en la prevalencia de obesidad/sobrepeso, dislipemia, resistencia a la insulina y alteración de la tensión arterial sistólica en supervivientes de leucemia aguda infantil a lo largo del tiempo, especialmente en aquellos con enfermedades y tratamientos más agresivos (AU)


Background: Survivors of childhood acute leukemia (AL) face an increased risk of metabolic and cardiovascular late effects which increase their long-term morbimotality. Objective: To assess the prevalence of obesity, insulinresistance, dyslipidemia and hypertension as cardiometabolic risk factors in survivors of a childhood AL, and also to determine possible causes for these adverse cardiometabolic traits. Material and methods: A retrospective cohort study of 47 pediatric acute leukemia survivors diagnosed between 0-15 years, with a ten years follow-up. Results: Forty percent of participants had at least one cardiometabolic risk factor. Dyslipidemia (increased LDL cholesterol) was the most frequent (38.3%), secondly overweight/obese (31.9%), followed by systolic hypertension (23.4%). Females in contrast to males had an increased risk of developing all three risk factors (RR 1.6; RR 3.16; RR 1.69; p < 0.05). Only 19.4% of participants developed insulin resistance, while none were diagnosed with diabetes mellitus. High risk acute leukemia survivors were significantly more likely than low risk leukemia survivors to manifest multiple cardiometabolic traits like overweight/obesity, insulin resitance and dyslipidemia (RR 3.56; RR 2.39; RR 2.53; p < 0.05). Also, those who received hematopoietic cell trasplantation had an increased prevalence of overweight/obesity, increased LDL-cholesterol and systolic hypertension. Radiotherapy treatment was also associated with insulin resitance and systolic hypertension (RR 2.47; RR 2.53; p < 0.05). Conclusions: There is an increased risk of overweight/obesity, dyslipidemia, insulin resistance and systolic blood pressure modifi cation in childhood acute leukemia survivors, specially in those who were diagnosed as a high risk acute laukemia and those treated with more aggressive treatments (AU)


Subject(s)
Humans , Male , Female , Child , Leukemia/epidemiology , Cardiovascular Diseases/epidemiology , Metabolic Syndrome/epidemiology , Survivors/statistics & numerical data , Obesity/epidemiology , Dyslipidemias/epidemiology , Retrospective Studies
11.
Immunobiology ; 221(1): 40-7, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26323380

ABSTRACT

Autoimmune lymphoproliferative syndrome (ALPS) is a primary immunodeficiency caused by impaired Fas/FasL-mediated apoptosis of lymphocytes and is characterized by chronic nonmalignant or benign lymphoproliferation, autoimmune manifestations and expansion of double negative (DN) T-cells (TCRαß+CD4-CD8-). Most cases of ALPS are associated with germline (ALPS-FAS) or somatic (ALPS-sFAS) heterozygous FAS mutations or a combination of both. Here we report three unrelated patients with ALPS-sFAS. Only one of them showed impaired Fas function in PHA-activated T-cells. In this patient, the genetic analysis of the caspase-10 gene (CASP10) identified a heterozygous germline change in exon 9 (c.1337A>G) causing Y446C substitution in the caspase-10 protein. In addition, this patient had a dysregulated T- and B-cell phenotype; circulating lymphocytes showed expansion of T effector memory CD45RA+ (TEMRA) CD4 T-cells, effector memory CD8 T-cells, CD21(low) B-cells and reduced memory switched B-cells. Additionally, this patient showed altered expression in T-cells of several molecules that change during differentiation from naïve to effector cells (CD27, CD95, CD57 and perforin). Molecular alterations in genes of the Fas pathway are necessary for the development of ALPS and this syndrome could be influenced by the concurrent effect of other mutations hitting different genes involved in Fas or related pathways.


Subject(s)
Autoimmune Lymphoproliferative Syndrome/genetics , B-Lymphocytes/immunology , CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Caspase 10/genetics , fas Receptor/genetics , Adolescent , Antigens, CD/genetics , Antigens, CD/immunology , Autoimmune Lymphoproliferative Syndrome/immunology , Autoimmune Lymphoproliferative Syndrome/pathology , B-Lymphocytes/pathology , CD4-Positive T-Lymphocytes/pathology , CD8-Positive T-Lymphocytes/pathology , Caspase 10/immunology , Exons , Female , Gene Expression , Humans , Immunologic Memory , Lymphatic Diseases/genetics , Lymphatic Diseases/immunology , Lymphatic Diseases/pathology , Lymphocyte Activation/drug effects , Male , Middle Aged , Mutation , Perforin/genetics , Perforin/immunology , Phytohemagglutinins/pharmacology , Primary Cell Culture , Splenomegaly/genetics , Splenomegaly/immunology , Splenomegaly/pathology , fas Receptor/immunology
12.
Med Phys ; 41(9): 092503, 2014 Sep.
Article in English | MEDLINE | ID: mdl-25186412

ABSTRACT

PURPOSE: 90Y-microspheres are widely used for the radioembolization of metastatic liver cancer or hepatocellular carcinoma and there is a growing interest for imaging 90Y-microspheres with PET. The aim of this study is to evaluate the performance of a current generation PET/CT scanner for 90Y imaging and to optimize the PET protocol to improve the assessment and the quantification of 90Y-microsphere biodistribution after radioembolization. METHODS: Data were acquired on a Biograph mCT-TrueV scanner with time of flight (TOF) and point spread function (PSF) modeling. Spatial resolution was measured with a 90Y point source. Sensitivity was evaluated using the NEMA 70 cm line source filled with 90Y. To evaluate the count rate performance, 90Y vials with activity ranging from 3.64 to 0.035 GBq were measured in the center of the field of view (CFOV). The energy spectrum was evaluated. Image quality with different reconstructions was studied using the Jaszczak phantom containing six hollow spheres (diameters: 31.3, 28.1, 21.8, 16.1, 13.3, and 10.5 mm), filled with a 207 kBq/ml 90Y concentration and a 5:1 sphere-to-background ratio. Acquisition time was adjusted to simulate the quality of a realistic clinical PET acquisition of a patient treated with SIR-Spheres®. The developed methodology was applied to ten patients after SIR-Spheres® treatment acquiring a 10 min per bed PET. RESULTS: The energy spectrum showed the 90Y bremsstrahlung radiation. The 90Y transverse resolution, with filtered backprojection reconstruction, was 4.5 mm in the CFOV and degraded to 5.0 mm at 10 cm off-axis. 90Y absolute sensitivity was 0.40 kcps/MBq in the center of the field of view. Tendency of true and random rates as a function of the 90Y activity could be accurately described using linear and quadratic models, respectively. Phantom studies demonstrated that, due to low count statistics in 90Y PET acquisition, the optimal parameters for the standard OSEM+PSF reconstruction were only one iteration and a postreconstruction filter of 6 mm FWHM, for both TOF and non-TOF reconstructions. Moreover, when TOF is included, the signal to noise ratio increased and visibility achieved 100% by the experienced observers and 93.3% according to the Rose model of statistical detection. In 50% of patients, TOF allowed the visualization of 90Y radioembolized lesions not seen without TOF, confirming phantom results. Liver activity was accurately quantified, with no significant differences between reconstructed and actual delivered activity to the whole-liver [mean relative difference (10.2±14.7)%]. CONCLUSIONS: Qualitative and quantitative 90Y PET imaging improved with the introduction of TOF in a PET/CT scanner, thereby allowing the visualization of microsphere deposition in lesions not visible in non-TOF images. This technique accurately quantifies the total activity delivered to the liver during radioembolization with (90)Y-microspheres and allows dose estimation.


Subject(s)
Brachytherapy , Embolization, Therapeutic , Microspheres , Positron-Emission Tomography/methods , Yttrium Radioisotopes/pharmacokinetics , Adenocarcinoma/diagnostic imaging , Adenocarcinoma/metabolism , Adenocarcinoma/therapy , Aged , Carcinoma, Hepatocellular/diagnostic imaging , Carcinoma, Hepatocellular/metabolism , Carcinoma, Hepatocellular/therapy , Cholangiocarcinoma/diagnostic imaging , Cholangiocarcinoma/metabolism , Cholangiocarcinoma/therapy , Colorectal Neoplasms/diagnostic imaging , Colorectal Neoplasms/metabolism , Colorectal Neoplasms/therapy , Computer Simulation , Esophageal Neoplasms/diagnostic imaging , Esophageal Neoplasms/metabolism , Esophageal Neoplasms/therapy , Humans , Liver/diagnostic imaging , Liver/metabolism , Liver Neoplasms/diagnostic imaging , Liver Neoplasms/metabolism , Liver Neoplasms/therapy , Male , Middle Aged , Neuroendocrine Tumors/diagnostic imaging , Neuroendocrine Tumors/metabolism , Neuroendocrine Tumors/therapy , Phantoms, Imaging , Positron-Emission Tomography/instrumentation , Radiation Dosage , Signal-To-Noise Ratio , Tissue Distribution , Yttrium Radioisotopes/therapeutic use
13.
J Immunol ; 187(11): 6130-42, 2011 Dec 01.
Article in English | MEDLINE | ID: mdl-22048768

ABSTRACT

Twenty-four patients with metastatic cancer received two cycles of four daily immunizations with monocyte-derived dendritic cells (DC). DC were incubated with preheated autologous tumor lysate and subsequently with IFN-α, TNF-α, and polyinosinic:polycytidylic acid to attain type 1 maturation. One DC dose was delivered intranodally, under ultrasound control, and the rest intradermally in the opposite thigh. Cyclophosphamide (day -7), GM-CSF (days 1-4), and pegIFN alpha-2a (days 1 and 8) completed each treatment cycle. Pretreatment with cyclophosphamide decreased regulatory T cells to levels observed in healthy subjects both in terms of percentage and in absolute counts in peripheral blood. Treatment induced sustained elevations of IL-12 in serum that correlated with the output of IL-12p70 from cultured DC from each individual. NK activity in peripheral blood was increased and also correlated with the serum concentration of IL-12p70 in each patient. Circulating endothelial cells decreased in 17 of 18 patients, and circulating tumor cells markedly dropped in 6 of 19 cases. IFN-γ-ELISPOT responses to DC plus tumor lysate were observed in 4 of 11 evaluated cases. Tracing DC migration with [(111)In] scintigraphy showed that intranodal injections reached deeper lymphatic chains in 61% of patients, whereas with intradermal injections a small fraction of injected DC was almost constantly shown to reach draining inguinal lymph nodes. Five patients experienced disease stabilization, but no objective responses were documented. This combinatorial immunotherapy strategy is safe and feasible, and its immunobiological effects suggest potential activity in patients with minimal residual disease. A randomized trial exploring this hypothesis is currently ongoing.


Subject(s)
Antigens, Neoplasm/immunology , Antineoplastic Agents/therapeutic use , Dendritic Cells/transplantation , Immunotherapy/methods , Neoplasms/therapy , Adjuvants, Immunologic/therapeutic use , Adult , Aged , Aged, 80 and over , Cancer Vaccines/immunology , Cancer Vaccines/therapeutic use , Cell Separation , Cyclophosphamide/therapeutic use , Dendritic Cells/immunology , Enzyme-Linked Immunosorbent Assay , Female , Flow Cytometry , Granulocyte-Macrophage Colony-Stimulating Factor/therapeutic use , Humans , Interferon-alpha/therapeutic use , Male , Middle Aged , Neoplasms/immunology , Pilot Projects , Polyethylene Glycols/therapeutic use , Recombinant Proteins/therapeutic use , T-Lymphocytes, Regulatory/drug effects , T-Lymphocytes, Regulatory/immunology
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