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Background: Long-term rhythm monitoring (LTRM) can detect undiagnosed atrial fibrillation (AF) in patients at risk of AF and stroke. Circulating microRNAs (miRNAs), which have been shown to play a role in atrial electrical and structural remodelling, could help to select patients who would benefit most from LTRM. The aim of this study was to investigate whether patients with diabetes mellitus (DM) and hypertension and screen-detected subclinical AF (SCAF) using an insertable cardiac monitor (ICM) have significantly different plasma baseline levels of five selected miRNAs playing a role in the modulation of atrial electrical and structural remodelling (miR-21-5p, miR-29b-3p, miR-150-5p, miR-328-3p, and miR-432-5p) compared to those without SCAF. Methods: This study was performed at the outpatient clinic of a secondary academic teaching hospital between December 2013 and November 2015. Eligible patients were ≥65 years of age with DM and hypertension but without known heart diseases. All patients received an ICM. On the day of ICM implantation, blood samples for the measurement of plasma levels of the five miRNAs were drawn. In this post hoc analysis, we investigated their expression by reverse transcription-quantitative polymerase chain reaction. MiRNA plasma levels in patients with and without newly detected SCAF were compared. Results: We included 82 consecutive patients (median age of 71.3 years (IQR 67.4-75.1)), who were followed for a median of 588 days (IQR: 453-712 days). Seventeen patients (20.7%) had ICM-detected SCAF. Plasma levels of miR-328-3p, miR-29b-3p, miR-21-5p, miR-432-5p, and miR-150-5p were slightly but not significantly different in patients with incident SCAF compared with patients without. Conclusions: In patients with hypertension and DM, newly detected SCAF was not significantly associated with changes in expression levels of miR-21-5p, miR-29b-3p, miR-150-5p, miR-328-3p, and miR-432-5p.
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After a positive phase III trial, it is evident that treatment with tumor-infiltrating lymphocytes (TIL) is a safe, feasible, and effective treatment modality for patients with metastatic melanoma. Further, the treatment is safe and feasible in diverse solid tumors, regardless of the histologic type. Still, TIL treatment has not obtained the regulatory approvals to be implemented on a larger scale. Therefore, its availability is currently restricted to a few centers worldwide. In this review, we present the current knowledge of TIL therapy and discuss the practical, logistic, and economic challenges associated with implementing TIL therapy on a larger scale. Finally, we suggest strategies to facilitate the widespread implementation of TIL therapy and approaches to develop the next generation of TILs.
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Inmunoterapia Adoptiva , Melanoma , Humanos , Linfocitos Infiltrantes de Tumor/patología , Melanoma/patologíaRESUMEN
BACKGROUND: Immune checkpoint inhibitors and targeted therapies have dramatically improved outcomes in patients with advanced melanoma, but approximately half these patients will not have a durable benefit. Phase 1-2 trials of adoptive cell therapy with tumor-infiltrating lymphocytes (TILs) have shown promising responses, but data from phase 3 trials are lacking to determine the role of TILs in treating advanced melanoma. METHODS: In this phase 3, multicenter, open-label trial, we randomly assigned patients with unresectable stage IIIC or IV melanoma in a 1:1 ratio to receive TIL or anti-cytotoxic T-lymphocyte antigen 4 therapy (ipilimumab at 3 mg per kilogram of body weight). Infusion of at least 5×109 TILs was preceded by nonmyeloablative, lymphodepleting chemotherapy (cyclophosphamide plus fludarabine) and followed by high-dose interleukin-2. The primary end point was progression-free survival. RESULTS: A total of 168 patients (86% with disease refractory to anti-programmed death 1 treatment) were assigned to receive TILs (84 patients) or ipilimumab (84 patients). In the intention-to-treat population, median progression-free survival was 7.2 months (95% confidence interval [CI], 4.2 to 13.1) in the TIL group and 3.1 months (95% CI, 3.0 to 4.3) in the ipilimumab group (hazard ratio for progression or death, 0.50; 95% CI, 0.35 to 0.72; P<0.001); 49% (95% CI, 38 to 60) and 21% (95% CI, 13 to 32) of the patients, respectively, had an objective response. Median overall survival was 25.8 months (95% CI, 18.2 to not reached) in the TIL group and 18.9 months (95% CI, 13.8 to 32.6) in the ipilimumab group. Treatment-related adverse events of grade 3 or higher occurred in all patients who received TILs and in 57% of those who received ipilimumab; in the TIL group, these events were mainly chemotherapy-related myelosuppression. CONCLUSIONS: In patients with advanced melanoma, progression-free survival was significantly longer among those who received TIL therapy than among those who received ipilimumab. (Funded by the Dutch Cancer Society and others; ClinicalTrials.gov number, NCT02278887.).
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Inmunoterapia Adoptiva , Linfocitos Infiltrantes de Tumor , Melanoma , Humanos , Tratamiento Basado en Trasplante de Células y Tejidos , Ipilimumab/efectos adversos , Melanoma/tratamiento farmacológicoRESUMEN
BACKGROUND: Long-term rhythm monitoring (LTRM) can detect undiagnosed atrial fibrillation (AF) in patients at high risk of AF and stroke. Biomarkers and echocardiographic parameters could, however, help identify patients benefitting most from LTRM. The aim of this study was to investigate, whether circulating biomarkers of cardiac and vascular function (brain natriuretic peptide (BNP), cardiac troponin I (cTnI), copeptin, and mid-regional proadrenomedullin (MR-proADM)) and echocardiographic parameters were associated with incident subclinical AF (SCAF) in a population at high risk of stroke in the presence of AF. For this purpose, we investigated individuals ≥65 years of age with hypertension and diabetes mellitus, but no history or symptoms of AF or other cardiovascular disease (CVD). METHODS: We included 82 consecutive patients (median age 71.3 years (IQR 67.4-75.1)). All patients received an insertable cardiac monitor (ICM) and were followed for a median of 588 days (IQR 453-712). On the day of ICM implantation, a comprehensive echocardiogram and blood samples were obtained. RESULTS: During a median follow-up of 588 days (IQR: 453-712 days), incident SCAF occurred in 17 patients (20.7%) with a median time to first-detected episode of 91 days (IQR 41-251 days). MR-proADM (median 0.87 nmol/L (IQR 0.76-1.02) vs 0.78 nmol/L (IQR 0.68-0.98)) and copeptin (median 13 pmol/L (IQR 9-17) vs 8 pmol/L (IQR 4-18)) levels were insignificantly higher in patients with incident SCAF. BNP and cTnI concentrations and echocardiographic parameters were similar in the two groups. CONCLUSIONS: MR-proADM, BNP, cTnI, copeptin, and several echocardiographic parameters were not associated with incident SCAF in this cohort of patients with hypertension and diabetes, but without any underlying CVD.
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Fibrilación Atrial/sangre , Fibrilación Atrial/diagnóstico por imagen , Biomarcadores/sangre , Complicaciones de la Diabetes/sangre , Complicaciones de la Diabetes/diagnóstico por imagen , Ecocardiografía , Hipertensión/complicaciones , Anciano , Electrocardiografía , Femenino , Humanos , Masculino , Estudios Prospectivos , Factores de RiesgoRESUMEN
An in-depth understanding of the impact of aging, cognitive reserve, and HIV status on cognitive function is needed in older West African adults. Ninety-nine HIV-negative and 334 HIV-positive adults aged ≥ 50 years were enrolled in three clinics (Senegal and Côte d'Ivoire) participating in the IeDEA West Africa collaboration. All subjects underwent the Free and Cued Selective Reminding Test (FCSRT) and the Isaacs Set Test (IST). Age (both linear and quadratic), education level, and HIV status effects on Z-scores were assessed using multivariate linear regression models. Interactions between HIV status and age or educational level were tested. In the present cohort of older West African adults, the role of age and educational level on episodic memory and verbal fluency was observed without revealing an interaction between HIV status and age effect. As age had quadratic effects, older HIV-positive adults should not be considered as a unique group irrespective of their age. Low-educated HIV-positive patients had the lowest verbal fluency performance compared to others. Further studies are needed to duplicate these results. In clinical settings, screening and adapted programs focusing on improving cognition in those patients are needed.
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Infecciones por VIH , Anciano , Cognición , Estudios de Cohortes , Côte d'Ivoire/epidemiología , Escolaridad , Infecciones por VIH/epidemiología , HumanosRESUMEN
OBJECTIVES: To cross-sectionally describe brain alterations in PLHIV aged above 50 years old, receiving antiretroviral treatment (ART) and living in Senegal compared to HIV-negative subjects. METHODS: Twenty PLHIV and 26 HIV-negative subjects with comparable socio-demographic and clinical characteristics underwent an MRI exam (3D-T1 and FLAIR sequences). Global atrophy and White Matter Hyperintensities (WMH) were evaluated. After assessing the feasibility and acceptability of MRI scans in this population, we described atrophy and WHM prevalence and associated factors using logistic regressions. RESULTS: Overall, 43.5% of the study sample were aged ≥60 years, 58.7% were women, and 28.3% had hypertension. The overall prevalence of atrophy and WMH was 19.6% [95% CI: 8.1-31.1] and 30.4% [95% CI: 17.1-43.7]. HIV status had no significant effect on atrophy or WMH. Unemployment and hypertension were significantly associated with atrophy, whereas women were less likely to present atrophy. Aged ≥60 years was the only factor associated with WMH. CONCLUSIONS: A high prevalence of atrophy and WMH was observed in West African adults aged over 50 years without a clear HIV impact. As brain MRI studies are critical to better understand cognitive and emotional outcomes, we encourage those studies in older PLHIV in West Africa.
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Encefalopatías/diagnóstico por imagen , Encefalopatías/etiología , Infecciones por VIH/complicaciones , Encéfalo/diagnóstico por imagen , Encéfalo/patología , Estudios Transversales , Femenino , Humanos , Hipertensión/complicaciones , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Prevalencia , SenegalRESUMEN
OBJECTIVES: The study sought to determine the incidence of subclinical atrial fibrillation (AF) in high-risk patients and to compare the effect of continuous versus intermittent monitoring. BACKGROUND: AF often occurs in a subclinical form, which makes it difficult to detect. The authors do not know the incidence of subclinical AF among patients ≥65 years of age with hypertension and diabetes mellitus. This group of patients has increased risk of developing AF and in addition a high thromboembolic risk, if AF is present. METHODS: A total of 82 outpatients ≥65 years of age (median age 71.3 years [interquartile range [IQR]: 67.4 to 75.1 years]) with hypertension and diabetes mellitus, and no history of AF or any other cardiovascular disease, were consecutively included. All patients received an insertable cardiac monitor (ICM) and were followed for a median of 588 days (IQR: 453 to 712 days). We compared continuous monitoring with 72-h Holter monitoring 1 month after ICM insertion. The primary endpoint was AF ≥2 min for the ICM and AF ≥30 s for the Holter monitoring. RESULTS: During follow-up 17 (20.7%) patients were found to have subclinical AF detected by ICM with a median time to first detected episode of 91 days (IQR: 41 to 251 days) from inclusion. Only 2 (2.4%) patients also had AF episodes on the 72-h Holter monitoring. All detected episodes were completely asymptomatic. CONCLUSIONS: The incidence of subclinical AF in this group of patients was surprisingly high. Continuous monitoring with ICM detected significantly more AF episodes than 72-h Holter monitoring. (Detection of Subclinical Atrial Fibrillation in High Risk Patients Using Implantable Loop Recorder; NCT02041832).
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Fibrilación Atrial/diagnóstico , Fibrilación Atrial/fisiopatología , Electrocardiografía Ambulatoria/métodos , Anciano , Fibrilación Atrial/epidemiología , Diabetes Mellitus/epidemiología , Electrocardiografía Ambulatoria/tendencias , Femenino , Estudios de Seguimiento , Humanos , Hipertensión/epidemiología , Incidencia , Masculino , Estudios Prospectivos , Factores de Riesgo , Tromboembolia/epidemiología , Tromboembolia/etiologíaRESUMEN
Introduction : La mortalité maternelle reste élevée dans la République Islamique de Mauritanie. L'Organisation Mondiale de la Santé recommande la promotion de la contraception moderne comme stratégie efficace de réduction de la réduction de la mortalité maternelle. L'objectif général était connaissances et pratiques des hommes en âge de procréer sur la contraception moderne dans la commune de Boghé, Mauritanie.Patients et méthodes : Il s'agissait d'une étude transversale, descriptive et analytique. La population d'étude était constituée par l'ensemble des hommes âgés d'au moins 18 ans vivants depuis au moins 3 mois dans de la commune de Boghé. Un sondage à deux degrés a été effectué. L'analyse multivariée et la régression logistique simple étaient utilisées grâce au logiciel R 2.2.9 pour identifier les caractéristiques qui influençaientla non-utilisation de la contraception moderne.Résultats : La taille de l'échantillon était de 384 hommes en âge de procréer. L'âge moyen des hommes en âge de procréer était de 42,1 ± 9,5 ans. Ils avaient entendu parler de la contraception moderne dans 70,3% des cas). Ils étaient informés par leurs parents/amis (76%), la radio (32%), leurs conjointes (18%), les services de santé (8%) et la télévision /journaux (5%). Les mariés, utilisaient une méthode contraceptive au sein du couple dans 9,4% des cas. Parmi eux, pour 80% leurs femmes utilisaient la pilule, un contraceptif injectable (14,3%) et des implants (5,7%). Le déterminant de la non-utilisation de méthodes contraceptives modernes était le manque d'informations relatives à la contraception moderne (OR ajusté = 8,13 (1,9-34,72)). Conclusion : Face à cette situation, il importe aux autorités sanitaires de Boghé de mener des activités de sensibilisation relatives à la contraception moderne tout en impliquant les hommes en vue de soutenir leurs conjointes à utuliser les produits contraceptifs modernes
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OBJECTIVE: To establish the respective prevalence of microalbuminuria and dyslipidemia and to evaluate their association with diabetes type 2. ANALYZE DATA: Prospective study of 195 type 2 diabetic subjects (125 women and 70 men) from a hospital in the city of Dakar (Senegal) for a check-up of diabetes. Age and sex were determined ; fasting blood glucose, glycated hemoglobin, total cholesterol, HDL cholesterol, LDL cholesterol, triglycerides and micro- albuminuria were measured. RESULTS: In this study, the mean age of patients was 57.9 ± 11.1 years. Age, glycated hemoglobin and microalbuminuria were significantly higher in women than in men (P < 0.01 ; P < 0.03 ; P < 0.01 respectively). The prevalence of microalbuminuria is 48.7% and that of dyslipidemia is 41.1%. Glycated hemoglobin is higher in subjects with microalbuminuria than in patients with normal microalbuminuria with a statistically significant difference (P < 0.001). There is a strong correlation (R = 0.82) between glycated hemoglobin and microalbuminuria, 1% increase in HbA1c corresponding approximately to an increase of 39.7 mg/I of microalbuminuria. CONCLUSION: Microalbuminuria and dyslipidemia are frequently found in type 2 diabetes, but the pathophysiological mechanisms of the association are not well known.
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Albuminuria/orina , Diabetes Mellitus Tipo 2/metabolismo , Dislipidemias/sangre , Adulto , Factores de Edad , Glucemia/análisis , Colesterol/sangre , HDL-Colesterol/sangre , LDL-Colesterol/sangre , Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/orina , Femenino , Hemoglobina Glucada/análisis , Humanos , Hipercolesterolemia/sangre , Hipertrigliceridemia/sangre , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Factores Sexuales , Triglicéridos/sangreRESUMEN
We have investigated prospectively how many pregnant women purchase ultrasound imaging before week 20, why they purchase scans, and which professional skills and certifications women expect the person performing the scan to have. In addition, we wanted to investigate whether the women were aware of any professional authorization procedures. Women attending the second trimester malformation scan of the Danish Prenatal Screening Program (n = 645) filled in a questionnaire about their use of non-medical ultrasound scans. Of these women, 154 (24%) had bought ultrasound scans: 50% wanted to have the fetal health and development checked and 49% wanted to find out the gender of the fetus. In addition, 68% felt that they received an evaluation of the fetal health state and 58% believed that there was legislation demanding a professional authorization needed to perform ultrasound imaging. This study shows that there is a significant demand among pregnant women for commercial ultrasound imaging of their fetus for various reasons. Knowledge of certifications and requirements for legal authorization is, however, sparse.
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Diagnóstico Prenatal/instrumentación , Diagnóstico Prenatal/estadística & datos numéricos , Autocuidado/métodos , Encuestas y Cuestionarios , Ultrasonografía Prenatal/estadística & datos numéricos , Adulto , Actitud Frente a la Salud , Estudios de Cohortes , Seguridad de Productos para el Consumidor , Dinamarca , Diagnóstico por Imagen/instrumentación , Diagnóstico por Imagen/estadística & datos numéricos , Femenino , Hospitales Universitarios/estadística & datos numéricos , Humanos , Incidencia , Embarazo , Segundo Trimestre del Embarazo , Estudios Prospectivos , Medición de Riesgo , Autocuidado/instrumentación , Transferencia de TecnologíaRESUMEN
A nonhuman primate model for malaria vaccine development allowing reliable, stringent sporozoite challenge and evaluation of both cellular and antibody responses is needed. We therefore constructed a multicomponent, multistage DNA vaccine for the simian malaria species Plasmodium knowlesi including two preerythrocytic-stage antigens, the circumsporozoite protein (PkCSP) and sporozoite surface protein 2 (PkSSP2), and two blood stage antigens, apical merozoite antigen 1 (PkAMA1) and merozoite surface protein 1 (PkMSP1p42), as well as recombinant canarypox viruses encoding the four antigens (ALVAC-4). The DNA vaccine plasmids expressed the corresponding antigens in vitro and induced antiparasite antibodies in mice. Groups of four rhesus monkeys received three doses of a mixture of the four DNA vaccine plasmids and a plasmid encoding rhesus granulocyte-monocyte colony-stimulating factor, followed by boosting with a single dose of ALVAC-4. Three groups received the priming DNA doses by different routes, either by intramuscular needle injection, by intramuscular injection with a needleless injection device, the Biojector, or by a combination of intramuscular and intradermal routes by Biojector. Animals immunized by any route developed antibody responses against sporozoites and infected erythrocytes and against a recombinant PkCSP protein, as well as gamma interferon-secreting T-cell responses against peptides from PkCSP. Following challenge with 100 P. knowlesi sporozoites, 1 of 12 experimental monkeys was completely protected and the mean parasitemia in the remaining monkeys was significantly lower than that in 4 control monkeys. This model will be important in preclinical vaccine development.
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Antígenos de Protozoos/inmunología , Avipoxvirus/genética , Vacunas contra la Malaria , Malaria/prevención & control , Plasmodium knowlesi/inmunología , Vacunas de ADN , Animales , Anticuerpos Antiprotozoarios/sangre , Antígenos de Protozoos/genética , Antígenos de Protozoos/metabolismo , Avipoxvirus/inmunología , Inmunización Secundaria/métodos , Interferón gamma/biosíntesis , Macaca mulatta , Vacunas contra la Malaria/administración & dosificación , Vacunas contra la Malaria/inmunología , Ratones , Parasitemia/prevención & control , Plásmidos/genética , Células Tumorales Cultivadas , Vacunas de ADN/administración & dosificación , Vacunas de ADN/inmunologíaRESUMEN
Antigens encoded by MAGE genes are of particular interest for cancer immunotherapy because of their strict tumoral specificity and because they are shared by many tumors. Antigenic peptide EVDPIGHLY encoded by MAGE-3 and known to be presented by HLA-A*0101 is currently being used in therapeutic vaccination trials. We report here that a cytolytic T-lymphocyte (CTL) clone, which is restricted by HLA-B*3501, recognizes the same peptide and, importantly, lyses HLA-B*3501 tumor cells expressing MAGE-3. These results infer that the current clinical use of peptide EVDPIGHLY can now be extended to HLA-B*3501 patients.
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Antígenos de Neoplasias , Antígeno HLA-A1/inmunología , Antígeno HLA-B35/inmunología , Proteínas de Neoplasias/inmunología , Linfocitos T Citotóxicos/inmunología , Secuencia de Aminoácidos , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Células Dendríticas/inmunología , Células Dendríticas/metabolismo , Epítopos/inmunología , Expresión Génica/inmunología , Humanos , Interferón gamma/metabolismo , Melanoma , Datos de Secuencia Molecular , Proteínas de Neoplasias/genética , Linfocitos T Citotóxicos/metabolismo , Transfección , Células Tumorales CultivadasRESUMEN
A reliable procedure to measure antigen specific T cell responses in rhesus macaques is required to determine the efficacy of vaccines and immunotherapies. The currently available T cell assays are poorly quantifiable or technically difficult to perform. Classical 51Cr-release cytotoxic T cell (CTL) assays are cumbersome and difficult to quantitate reproducibly. Detection of specific T-cell using MHC-peptide tetrameric complexes is highly sensitive, but requires knowledge of MHC type and prior identification of T cell epitopes. We therefore developed a rhesus interferon-gamma (IFN-gamma) ELISPOT assay capable of detecting IFN-gamma secretion in response to stimulation with pooled 20-mer peptides. Peripheral blood mononuclear cells (PBMCs) from rhesus monkeys immunized with a DNA vaccine and recombinant canary pox encoding the Plasmodium knowlesi circumsporozoite protein (PkCSP) were incubated with pools of peptides from PkCSP. Positive responses to peptide pools and individual peptides ranging from 100 to 450 spot forming cells (SFC)/10(6) PBMC were detected in four of four immunized monkeys and in zero of two control monkeys. In two monkeys studied in detail, the IFN-gamma response was focussed on a single 20-mer peptide, QGDGANAGQPQAQGDGANAG, and was dependent on CD4(+), but not CD8(+), T cells. Background responses in control monkeys and preimmunization PBMCs ranged from 10 to 50 SFC/10(6) PBMC. The average within assay and between assay coefficients of variation (CV) for this peptide ELISPOT were 21.9 and 24.7%, respectively. This peptide IFN-gamma assay will be a useful tool for evaluation of T cell responses in rhesus macaques.
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Antígenos de Protozoos/inmunología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Interferón gamma/inmunología , Vacunas contra la Malaria/inmunología , Secuencia de Aminoácidos , Animales , Criopreservación , Femenino , Técnicas para Inmunoenzimas/métodos , Interferón gamma/biosíntesis , Leucocitos Mononucleares/inmunología , Macaca mulatta , Masculino , Datos de Secuencia Molecular , Péptidos/inmunología , Plasmodium knowlesi/inmunología , Proteínas Protozoarias/inmunología , Reproducibilidad de los ResultadosRESUMEN
HLA-A*0201-restricted CTL against human gp100 were isolated from HLA-A*0201/K(b) (A2/K(b))-transgenic mice immunized with recombinant canarypox virus (ALVAC-gp100). These CTL strongly responded to the gp100(154-162) epitope, in the context of both the chimeric A2/K(b) and the wild-type HLA-A*0201- molecule, and efficiently lysed human HLA-A*0201(+), gp100(+) melanoma cells in vitro. The capacity of the CTL to eradicate these tumors in vivo was analyzed in A2/K(b)-transgenic transgenic mice that had received a tumorigenic dose of human uveal melanoma cells in the anterior chamber of the eye. This immune-privileged site offered the unique opportunity to graft xenogeneic tumors into immunocompetent A2/K(b)-transgenic mice, a host in which they otherwise would not grow. Importantly, systemic (i.v.) administration of the A2/K(b)-transgenic gp100(154-162)-specific CTL resulted in rapid elimination of the intraocular uveal melanomas, indicating that anti-tumor CTL are capable of homing to the eye and exerting their tumoricidal effector function. Flow cytometry analysis of ocular cell suspensions with HLA-A*0201-gp100(154-162) tetrameric complexes confirmed the homing of adoptively transferred CTL. Therefore, the immune-privileged state of the eye permitted the outgrowth of xenogeneic uveal melanoma cells, but did not protect these tumors against adoptive immunotherapy with highly potent anti-tumor CTL. These data constitute the first direct indication that immunotherapy of human uveal melanoma may be feasible.
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Marcación de Gen , Inmunoterapia Adoptiva/métodos , Melanoma/inmunología , Melanoma/terapia , Glicoproteínas de Membrana/genética , Proteínas de Neoplasias/genética , Linfocitos T Citotóxicos/trasplante , Neoplasias de la Úvea/inmunología , Neoplasias de la Úvea/terapia , Animales , Vacunas contra el Cáncer/genética , Vacunas contra el Cáncer/inmunología , Células Clonales , Pruebas Inmunológicas de Citotoxicidad , Citotoxicidad Inmunológica/genética , Epítopos de Linfocito T/inmunología , Antígenos H-2/genética , Antígeno HLA-A2/genética , Humanos , Activación de Linfocitos/genética , Melanoma/genética , Melanoma/patología , Glicoproteínas de Membrana/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Proteínas de Neoplasias/inmunología , Linfocitos T Citotóxicos/inmunología , Células Tumorales Cultivadas/trasplante , Neoplasias de la Úvea/genética , Vacunas Virales/genética , Vacunas Virales/inmunología , Antígeno gp100 del MelanomaRESUMEN
The generation of cytotoxic effector T cells requires delivery of two signals, one derived from a specific antigenic epitope and one from a costimulatory molecule. A phase I clinical trial was conducted with a non-replicating canarypoxvirus (ALVAC) constructed to express both human carcinoembryonic antigen (CEA) and the B7.1 costimulatory molecule. This was the first study in cancer patients to determine if the delivery of costimulation with a tumor vaccine was feasible and improved immune responses. Three cohorts of six patients, each with advanced CEA-expressing adenocarcinomas, were treated with increasing doses of an ALVAC-CEA-B7.1 vaccine (4.5 x 10(6), 4.5 x 10(7), and 4.5 x 10(8) plaque-forming units, PFU). Patients were vaccinated by intramuscular injection every 4 weeks for 3 months and monitored for side-effects, tumor growth and anti-CEA immune responses. ALVAC-CEA-B7.1 at doses up to 4.5 x 10(8) PFU was given without evidence of significant toxicity or autoimmune reactions. Three patients experienced clinically stable disease that correlated with increasing CEA-specific precursor T cells, as shown by in vitro interferon-gamma enzyme-linked immunoassay spot tests (ELISPOT). These three patients underwent repeated vaccination resulting in augmented CEA-specific T cell responses. This study represents the first use of costimulation to enhance antitumor vaccines in cancer patients. This approach resulted in CEA-specific immunity associated with stable diseases in three patients. This study also demonstrated that CEA-specific T cell responses could be sustained by repeated vaccinations. Although the number of patients was small, the addition of B7.1 to virus-based vaccines may improve immunological and stable diseases to vaccination against tumor-associated antigens with tolerable toxicity.
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Avipoxvirus/genética , Avipoxvirus/inmunología , Antígeno B7-1/genética , Vacunas contra el Cáncer/uso terapéutico , Antígeno Carcinoembrionario/genética , Adenocarcinoma/inmunología , Adenocarcinoma/terapia , Adulto , Anciano , Antígeno B7-1/inmunología , Vacunas contra el Cáncer/inmunología , Vacunas contra el Cáncer/toxicidad , Antígeno Carcinoembrionario/inmunología , Línea Celular , Células Cultivadas , Estudios de Cohortes , Neoplasias del Colon/inmunología , Neoplasias del Colon/terapia , Relación Dosis-Respuesta a Droga , Ensayo de Inmunoadsorción Enzimática , Epítopos/metabolismo , Femenino , Citometría de Flujo , Humanos , Interferón gamma/inmunología , Interferón gamma/metabolismo , Neoplasias Pulmonares/inmunología , Neoplasias Pulmonares/terapia , Masculino , Persona de Mediana Edad , Neoplasias Pancreáticas/inmunología , Neoplasias Pancreáticas/terapia , Péptidos/uso terapéutico , Fenotipo , Neoplasias del Recto/inmunología , Neoplasias del Recto/terapia , Neoplasias Gástricas/inmunología , Neoplasias Gástricas/terapia , Linfocitos T/inmunología , Factores de TiempoRESUMEN
Antigens encoded by melanoma antigen (MAGE) genes are of particular interest for cancer immunotherapy because of their strict tumoral specificity and because they are shared by many tumors. Antigenic peptide EADPTGHSY encoded by MAGE-A1 and known to be presented by HLA-A1 is currently being used in therapeutic vaccination trials. We report here that a cytotoxic T-lymphocyte (CTL) clone, which is restricted by HLA-B35, recognizes the same peptide and, importantly, lyses HLA-B35 tumor cells expressing MAGE-A1. This peptide can be presented to CTL by both HLA-B*3501 and HLA-B*3503 molecules, which are expressed by approximately 19% of Caucasians. These results infer that the current clinical use of peptide EADPTGHSY can now be extended to HLA-B35 patients.
Asunto(s)
Presentación de Antígeno , Antígeno HLA-A1/inmunología , Antígeno HLA-B35/inmunología , Melanoma/inmunología , Proteínas de Neoplasias/inmunología , Linfocitos T Citotóxicos/inmunología , Animales , Antígenos de Neoplasias , Células COS , Pruebas Inmunológicas de Citotoxicidad , Humanos , Melanoma/genética , Antígenos Específicos del Melanoma , Proteínas de Neoplasias/genética , Péptidos/inmunología , Transfección , Células Tumorales CultivadasRESUMEN
Intramuscular immunization with a naked DNA plasmid expressing the Plasmodium yoelii circumsporozoite protein (pPyCSP) protects mice against challenge with P. yoelii sporozoites. This protection can be improved either by coadministration of a plasmid expressing murine GM-CSF (pGMCSF) or by boosting with recombinant poxvirus expressing the PyCSP. We now report that combining these two strategies, by first mixing the priming dose of pPyCSP with pGMCSF and then boosting with recombinant virus, can substantially increase vaccine effectiveness. Not only were immune responses and protection improved but the pPyCSP dose could be lowered from 100 microg to 1 microg with little loss of immunogenicity after boost with recombinant poxvirus. Comparing mice primed by the 1-microg doses of pPyCSP plus 1 microg pGMCSF with mice primed by 1-microg doses of pPyCSP alone, the former were better protected (60% vs 0) and had higher concentrations of Abs (titers of 163, 840 vs 5, 120 by indirect fluorescent Ab test against sporozoites), more ex vivo CTL activity (25% vs 7% specific lysis), and more IFN-gamma-secreting cells by enzyme-linked immunospot assay (1460 vs 280 IFN-gamma spot-forming cells/106 cells). Priming with plasmid vaccine plus pGMCSF and boosting with recombinant poxviruses strongly improves the immunogenicity and protective efficacy of DNA vaccination and allows for significant reduction of dose.
Asunto(s)
Antígenos de Protozoos/inmunología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Vacunas contra la Malaria/inmunología , Plásmidos/inmunología , Vacunas de ADN/inmunología , Virus Vaccinia/genética , Vacunas Virales/inmunología , Animales , Anticuerpos Antiprotozoarios/biosíntesis , Antígenos de Protozoos/administración & dosificación , Antígenos de Protozoos/genética , Células Cultivadas , Relación Dosis-Respuesta Inmunológica , Epítopos de Linfocito T/inmunología , Femenino , Factor Estimulante de Colonias de Granulocitos y Macrófagos/administración & dosificación , Factor Estimulante de Colonias de Granulocitos y Macrófagos/inmunología , Antígenos H-2/inmunología , Inmunización Secundaria , Malaria/inmunología , Malaria/prevención & control , Vacunas contra la Malaria/administración & dosificación , Vacunas contra la Malaria/genética , Ratones , Ratones Endogámicos A , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Plásmidos/administración & dosificación , Plasmodium yoelii/genética , Plasmodium yoelii/inmunología , Proteínas Protozoarias/administración & dosificación , Proteínas Protozoarias/genética , Proteínas Protozoarias/inmunología , Linfocitos T/inmunología , Linfocitos T/metabolismo , Linfocitos T/parasitología , Vacunas de ADN/administración & dosificación , Virus Vaccinia/inmunología , Vacunas Virales/administración & dosificación , Vacunas Virales/genéticaRESUMEN
DNA vaccine plasmids were constructed that encoded four pre-erythrocytic antigens from the human malaria parasite Plasmodium falciparum: circumsporozoite protein (PfCSP); sporozoite surface protein 2 (PfSSP2); carboxyl terminus of liver stage antigen 1 (PfLSA-1 c-term); and, exported protein 1 (PfExp-1). Antigen expression was evaluated in vitro by immunoblot analysis of tissue culture cells following transient transfection with each plasmid. Clearly detectable levels of expression depended upon, or were markedly enhanced by, fusion of the antigen encoding sequences in-frame with the initiation complex and peptide leader sequence of human tissue plasminogen activator protein. Mice injected with these plasmids produced antigen specific antibody and cytotoxic T lymphocyte responses. However, the magnitudes of the responses were not always predicted by the in vitro expression assay. The results of this study provided the basis for further testing of these plasmids in primates and the formulation of multi-component pre-erythrocytic DNA vaccines for efficacy testing in human volunteers.
Asunto(s)
ADN Protozoario/inmunología , Vacunas contra la Malaria/genética , Vacunas contra la Malaria/inmunología , Plasmodium falciparum/genética , Vacunas de ADN/inmunología , Secuencia de Aminoácidos , Animales , Anticuerpos Antiprotozoarios/inmunología , Antígenos de Protozoos/biosíntesis , Antígenos de Protozoos/genética , Antígenos de Protozoos/inmunología , ADN Protozoario/genética , Humanos , Vacunas contra la Malaria/uso terapéutico , Malaria Falciparum/inmunología , Malaria Falciparum/prevención & control , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Plásmidos/genética , Linfocitos T Citotóxicos/inmunología , Vacunas de ADN/genética , Vacunas de ADN/uso terapéuticoRESUMEN
CD8(+) T cells have been implicated as critical effector cells in protective immunity against malaria parasites developing within hepatocytes. A vaccine that protects against malaria by inducing CD8(+) T cells will probably have to include multiple epitopes on the same protein or different proteins, because of parasite polymorphism and genetic restriction of T-cell responses. To determine if CD8(+) T-cell responses against multiple P. falciparum proteins can be induced in primates by immunization with plasmid DNA, rhesus monkeys were immunized intramuscularly with a mixture of DNA plasmids encoding four P. falciparum proteins or with individual plasmids. All six monkeys immunized with PfCSP DNA, seven of nine immunized with PfSSP2 DNA, and five of six immunized with PfExp-1 or PfLSA-1 DNA had detectable antigen-specific cytotoxic T lymphocytes (CTL) after in vitro restimulation of peripheral blood mononuclear cells. CTL activity was genetically restricted and dependent on CD8(+) T cells. By providing the first evidence for primates that immunization with a mixture of DNA plasmids induces CD8(+) T-cell responses against all the components of the mixture, these studies provide the foundation for multigene immunization of humans.