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Artisanal and small-scale gold mining (ASGM) is the primary global source of anthropogenic mercury (Hg) emissions. It has impacted the Amazon rainforest in the Peruvian region of Madre de Dios. However, few studies have investigated Hg's distribution in terrestrial ecosystems in this region. We studied Hg's distribution and its predictors in soil and native plant species from artisanal mining sites. Total Hg concentrations were determined in soil samples collected at different depths (0-5 cm and 5-30 cm) and plant samples (roots, shoots, leaves) from 19 native plant species collected in different land cover categories: naked soil (L1), gravel piles (L2), natural regeneration (L3), reforestation (L4), and primary forest (L5) in the mining sites. Hg levels in air were also studied using passive air samplers. The highest Hg concentrations in soil (average 0.276 and 0.210 mg kg-1 dw.) were found in the intact primary forest (L5) at 0-5 cm depth and in the plant rooting zones at 5-30 cm depth, respectively. Moreover, the highest Hg levels in plants (average 0.64 mg kg-1 dw) were found in foliage of intact primary forest (L5). The results suggest that the forest in these sites receives Hg from the atmosphere through leaf deposition and that Hg accumulates in the soil surrounding the roots. The Hg levels found in the plant leaves of the primary forest are the highest ever recorded in this region, exceeding values found in forests impacted by Hg pollution worldwide and raising concerns about the extent of the ASGM impact in this ecosystem. Correlations between Hg concentrations in soil, bioaccumulation in plant roots, and soil physical-chemical characteristics were determined. Linear regression models showed that the soil organic matter content (SOM), pH, and electrical conductivity (EC) predict the Hg distribution and accumulation in soil and bioaccumulation in root plants.
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Monitoreo del Ambiente , Oro , Mercurio , Minería , Contaminantes del Suelo , Suelo , Mercurio/análisis , Perú , Contaminantes del Suelo/análisis , Suelo/química , Plantas/metabolismo , Ecosistema , Bosques , Bosque LluviosoRESUMEN
Artisanal and Small-Scale Gold Mining (ASGM) represents a significant source of anthropogenic mercury emissions to the environment, with potentially severe implications for avian biodiversity. In the Madre de Dios department of the southern Peruvian Amazon, ASGM activities have created landscapes marred by deforestation and post-mining water bodies (mining ponds) with notable methylation potential. While data on Hg contamination in terrestrial wildlife remains limited, this study measures Hg exposure in several terrestrial bird species as bioindicators. Total Hg (THg) levels in feathers from birds near water bodies, including mining ponds associated with ASGM areas and oxbow lakes, were analyzed. Our results showed significantly higher Hg concentrations in birds from ASGM sites with mean ± SD of 3.14 ± 7.97 µg/g (range: 0.27 to 72.75 µg/g, n = 312) compared to control sites with a mean of 0.47 ± 0.42 µg/g (range: 0.04 to 1.89 µg/g, n = 52). Factors such as trophic guilds, ASGM presence, and water body area significantly influenced feather Hg concentrations. Notably, piscivorous birds exhibited the highest Hg concentration (31.03 ± 25.25 µg/g, n = 12) exceeding known concentrations that affect reproductive success, where one measurement of Chloroceryle americana (Green kingfisher; 72.7 µg/g) is among the highest ever reported in South America. This research quantifies Hg exposure in avian communities in Amazonian regions affected by ASGM, highlighting potential risks to regional bird populations.
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Aves , Monitoreo del Ambiente , Oro , Mercurio , Minería , Animales , Mercurio/análisis , Perú , Plumas/química , Contaminantes Químicos del Agua/análisis , Contaminantes Ambientales/análisis , Exposición a Riesgos AmbientalesRESUMEN
Mercury (Hg) endangers human and wildlife health globally, primarily due to its release from artisanal small-scale gold mining (ASGM). During gold extraction, Hg is emitted into the environment and converted to highly toxic methylmercury by microorganisms. In Madre de Dios (MDD), Peru, ASGM dominates the economy and has transformed rainforests into expansive deforested areas punctuated by abandoned mining ponds. The aim of this study was to evaluate the use of bats as bioindicators for mercury pollution intensity in tropical terrestrial ecosystems impacted by ASGM. We collected 290 bat fur samples from three post-ASGM sites and one control site in Madre de Dios. Our results showed a wide Hg distribution in bats (0.001 to 117.71 mg/kg) strongly influenced by feeding habits. Insectivorous and piscivorous bats from ASGM sites presented elevated levels of Hg surpassing the mercury small mammal threshold for small mammals (10 mg/kg). We observed the highest reported fur mercury concentrations for insectivorous Neotropical bats reported to date (Rhynchonycteris naso, 117 mg/kg). Our findings further confirm that Hg emissions from ASGM are entering local food webs and exposing wildlife species at several trophic levels to higher levels of Hg than in areas not impacted by mining. We also found that three bat genera consistently showed increased Hg levels in ASGM sites relative to controls indicating potential usefulness as bioindicators of mercury loading in terrestrial ecosystems impacted by artisanal and small-scale gold mining.
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Bioacumulación , Quirópteros , Ecosistema , Monitoreo del Ambiente , Oro , Mercurio , Minería , Animales , Quirópteros/metabolismo , Perú , Mercurio/análisis , Mercurio/metabolismo , Monitoreo del Ambiente/métodos , Contaminantes Ambientales/metabolismo , Contaminantes Ambientales/análisisRESUMEN
The Matsigenka people living traditional lifestyles in remote areas of the Amazon rely on a fish-based diet that exposes them to methylmercury (MeHg) at levels that have been associated with decreased IQ scores. In this study, the association between Hg levels and working memory was explored using the framework of the Multicomponent Model. Working memory tasks were modified to fit the culture and language of the Matsigenka when needed and included measures for verbal storage (Word Span) visuospatial storage (Corsi Block Task) and a measure of executive functions, the Self-Ordered Pointing Task (SOPT). An innovation of the Trail Making Tests A & B (TMT A & B) was pilot tested as another potential measure of executive functions. The mean hair Hg levels of 30 participants, ages 12 to 55 years, from three different communities (Maizal, Cacaotal and Yomibato) was 7.0 ppm (sd = 2.40), well above the World Health Organization (WHO) limit for hair of 2.0 ppm and ranged from 1.8 to 14.2 ppm, with 98% of a broader sample of 152 individuals exceeding the WHO limit. Hair Hg levels showed significant associations with cognitive performance, but the degree varied in magnitude according to the type of task. Hg levels were negatively associated with executive functioning performance (SOPT errors), while Hg levels and years of education predicted visuospatial performance (Corsi Block accuracy). Education was the only predictor of Word Span accuracy. The results show that Hg exposure is negatively associated with working memory performance when there is an increased reliance on executive functioning. Based on our findings and the review of the experimental research, we suggest that the SOPT and the Corsi Block have the potential to be alternatives to general intelligence tests when studying remote groups with extensive cultural differences.
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Memoria a Corto Plazo , Mercurio , Animales , Función Ejecutiva , Humanos , Pueblos Indígenas , Mercurio/análisis , Pruebas Neuropsicológicas , PerúRESUMEN
Artisanal and small-scale gold mining (ASGM) is the primary global source of anthropogenic mercury (Hg) emissions and a large source of landscape change. ASGM occurs throughout the world, including in the Peruvian Amazon. This data set contains measurements of surface water, precipitation, throughfall, leaves, sediment, soil, and air samples from across the Madre de Dios region of Peru, in locations near and remote from ASGM. These data were collected to determine the fate and transport of Hg across the landscape. Samples were collected in 2018 and 2019. Data predominantly included total Hg and methyl Hg concentrations in surface water, precipitation, throughfall, leaves, sediment, soil, and air. Additional water and soil parameters were also measured to better characterize their chemistry. There are no copyright restrictions; please cite this data paper when the data are used in publication.
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Mercurio , Suelo , Monitoreo del Ambiente , Oro , Mercurio/análisis , Minería , Perú , Hojas de la Planta/química , AguaRESUMEN
Mercury emissions from artisanal and small-scale gold mining throughout the Global South exceed coal combustion as the largest global source of mercury. We examined mercury deposition and storage in an area of the Peruvian Amazon heavily impacted by artisanal gold mining. Intact forests in the Peruvian Amazon near gold mining receive extremely high inputs of mercury and experience elevated total mercury and methylmercury in the atmosphere, canopy foliage, and soils. Here we show for the first time that an intact forest canopy near artisanal gold mining intercepts large amounts of particulate and gaseous mercury, at a rate proportional with total leaf area. We document substantial mercury accumulation in soils, biomass, and resident songbirds in some of the Amazon's most protected and biodiverse areas, raising important questions about how mercury pollution may constrain modern and future conservation efforts in these tropical ecosystems.
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Atmósfera/análisis , Contaminantes Ambientales/análisis , Oro , Mercurio/análisis , Minería , Bosque Lluvioso , Animales , Atmósfera/química , Aves/clasificación , Aves/metabolismo , Monitoreo del Ambiente/métodos , Contaminación Ambiental/análisis , Plumas/química , Gases/análisis , Geografía , Perú , Hojas de la Planta/química , Hojas de la Planta/metabolismo , Suelo/química , Contaminantes del Suelo/análisis , Contaminantes del Suelo/metabolismoRESUMEN
Artificial marker mapping is a useful tool for fast camera localization estimation with a certain degree of accuracy in large indoor and outdoor environments. Nonetheless, the level of accuracy can still be enhanced to allow the creation of applications such as the new Visual Odometry and SLAM datasets, low-cost systems for robot detection and tracking, and pose estimation. In this work, we propose to improve the accuracy of map construction using artificial markers (mapping method) and camera localization within this map (localization method) by introducing a new type of artificial marker that we call the smart marker. A smart marker consists of a square fiducial planar marker and a pose measurement system (PMS) unit. With a set of smart markers distributed throughout the environment, the proposed mapping method estimates the markers' poses from a set of calibrated images and orientation/distance measurements gathered from the PMS unit. After this, the proposed localization method can localize a monocular camera with the correct scale, directly benefiting from the improved accuracy of the mapping method. We conducted several experiments to evaluate the accuracy of the proposed methods. The results show that our approach decreases the Relative Positioning Error (RPE) by 85% in the mapping stage and Absolute Trajectory Error (ATE) by 50% for the camera localization stage in comparison with the state-of-the-art methods present in the literature.
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Recent studies from the Madre de Dios region of Peru report elevated mercury exposure among the general population. Our objective was to assess mercury exposure, risk factors, risk perceptions, and knowledge among women of childbearing age, a population of concern due to risks to infant and fetal health. We collected hair samples and administered a survey among 200 women aged 18-49 years residing in Madre de Dios. Hair total mercury (THg) concentrations were analyzed in accordance with EPA Method 7473. Associations between exposures and potential risk factors were analyzed using generalized linear mixed models. Geometric mean hair THg concentration (±standard deviation) for all participants was 1.62⯱â¯2.54⯵g/g (range: 0.01-30.12⯵g/g), and 67 participants (33.5%) had concentrations above the 2.2⯵g/g reference dose established by the World Health Organization (WHO). Higher exposure was significantly associated with higher fish consumption (pâ¯=â¯0.03) and less mercury knowledge (pâ¯<â¯0.0001), but not with age or educational attainment. Most participants (86.2%) expressed concern about mercury contamination, but few (26.2%) had knowledge of the health risks associated with exposure. Many residents consumed fish species previously found to have low mercury concentrations, but more than 25% of participants reported consuming fish species with high mercury concentrations. We did not observe cross-sectional differences in fish intake among women who were pregnant or breastfeeding at the time of data collection.
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Exposición a Riesgos Ambientales/estadística & datos numéricos , Oro , Mercurio , Minería , Adolescente , Adulto , Animales , Estudios Transversales , Femenino , Peces , Contaminación de Alimentos , Humanos , Persona de Mediana Edad , Perú , Factores de Riesgo , Adulto JovenRESUMEN
Cross-presentation is an important mechanism for the differentiation of effector cytotoxic T lymphocytes (CTL) from naïve CD8+ T-cells, a key response for the clearance of intracellular pathogens and tumors. The liposomal co-encapsulation of the pore-forming protein sticholysin II (StII) with ovalbumin (OVA) (Lp/OVA/StII) induces a powerful OVA-specific CTL activation and an anti-tumor response in vivo. However, the pathway through which the StII contained in this preparation is able to induce antigen cross-presentation and the type of professional antigen presenting cells (APCs) involved have not been elucidated. Here, the ability of mouse bone marrow-derived dendritic cells (BM-DCs) and macrophages (BM-MΦs) stimulated with Lp/OVA/StII to activate SIINFEKL-specific B3Z CD8+ T cells was evaluated in the presence of selected inhibitors. BM-MΦs, but not BM-DCs were able to induce SIINFEKL-specific B3Z CD8+ T cell activation upon stimulation with Lp/OVA/StII. The cross-presentation of OVA was markedly decreased by the lysosome protease inhibitors, leupeptin and cathepsin general inhibitor, while it was unaffected by the proteasome inhibitor epoxomicin. This process was also significantly reduced by phagocytosis and Golgi apparatus function inhibitors, cytochalasin D and brefeldin A, respectively. These results are consistent with the concept that BM-MΦs internalize these liposomes through a phagocytic mechanism resulting in the cross-presentation of the encapsulated OVA by the vacuolar pathway. The contribution of macrophages to the CTL response induced by Lp/OVA/StII in vivo was determined by depleting macrophages with clodronate-containing liposomes. CTL induction was almost completely abrogated in mice depleted of macrophages, demonstrating the relevance of these APCs in the antigen cross-presentation induced by this formulation.
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Venenos de Cnidarios/metabolismo , Células Dendríticas/fisiología , Macrófagos/fisiología , Linfocitos T Citotóxicos/inmunología , Vacuolas/metabolismo , Animales , Antígenos/inmunología , Antígenos CD8/metabolismo , Células Cultivadas , Venenos de Cnidarios/química , Reactividad Cruzada , Femenino , Leupeptinas/farmacología , Liposomas/química , Activación de Linfocitos , Ratones , Ratones Endogámicos C57BL , Ovalbúmina/inmunologíaRESUMEN
Numerous molecules have been considered as targets for cancer immunotherapy because of their levels of expression on tumor cells, their putative importance for tumor biology, and relative immunogenicity. In this review we focus on the ganglioside GM3(Neu5Gc), a glycosphingolipid present on the outer side of the plasma membrane of vertebrate cells. The reasons for selecting GM3(Neu5Gc) as a tumor-specific antigen and its use as a target for cancer immunotherapy are discussed, together with the development of antitumor therapies focused on this target by the Center of Molecular Immunology (CIM, Cuba).
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Anticuerpos Monoclonales/inmunología , Antígenos de Neoplasias/inmunología , Gangliósido G(M3)/inmunología , Neoplasias/inmunología , Animales , Anticuerpos Monoclonales/uso terapéutico , Anticuerpos Monoclonales de Origen Murino , Antígenos de Neoplasias/química , Antígenos de Neoplasias/metabolismo , Secuencia de Carbohidratos , Modelos Animales de Enfermedad , Gangliósido G(M3)/antagonistas & inhibidores , Gangliósido G(M3)/química , Humanos , Inmunoterapia/métodos , Neoplasias/tratamiento farmacológico , Neoplasias/metabolismoRESUMEN
Artisanal and small-scale gold mining (ASGM) in Madre de Dios, Peru, continues to expand rapidly, raising concerns about increases in loading of mercury (Hg) to the environment. We measured physicochemical parameters in water and sampled and analyzed sediments and fish from multiple sites along one ASGM-impacted river and two unimpacted rivers in the region to examine whether Hg concentrations were elevated and possibly related to ASGM activity. We also analyzed the 308 fish samples, representing 36 species, for stable isotopes (δ15N and δ13C) to estimate their trophic position. Trophic position was positively correlated with the log-transformed Hg concentrations in fish among all sites. There was a lack of relationship between Hg concentrations in fish and either Hg concentrations in sediments or ASGM activity among sites, suggesting that fish Hg concentrations alone is not an ideal bioindicator of site-specific Hg contamination in the region. Fish Hg concentrations were not elevated in the ASGM-impacted river relative to the other two rivers; however, sediment Hg concentrations were highest in the ASGM-impacted river. Degraded habitat conditions and commensurate shifts in fish species and ecological processes may influence Hg bioaccumulation in the ASGM-impacted river. More research is needed on food web dynamics in the region to elucidate any effects caused by ASGM, especially through feeding relationships and food sources.
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Peces/metabolismo , Oro , Mercurio/análisis , Minería/estadística & datos numéricos , Ríos/química , Animales , Ecosistema , Monitoreo del Ambiente , Cadena Alimentaria , Perú , Contaminantes Químicos del Agua/análisisRESUMEN
This study aimed to investigate the immunogenicity of a cancer vaccine consisting of the NeuGcGM3 ganglioside combined with the outer membrane protein complex of Neisseria meningitides to form very small size particles. The vaccine is administered together with Montanide ISA51, as adjuvant treatment for breast cancer patients. After surgical resection and standard first-line chemo/radiotherapy, breast cancer patients in stage II-III were enrolled in a phase III clinical trial and allocated into 2 strata, according to the number of positive lymph nodes [stratum I (0-3); stratum II (≥4)]. Subsequently, patients were randomly assigned to receive the vaccine or placebo. The treatment consisted of 5 vaccine doses (200 µg) every 2 weeks and thereafter monthly reimmunizations to complete 15 doses. The vaccine was well-tolerated and high titers of immunoglobulin M and immunoglobulin G anti-NeuGcGM3 antibodies were similarly detected in each stratum. Hyperimmune sera were able to specifically recognize and kill the NeuGcGM3-expressing L1210 tumor cell line, and these functional capacities were significantly associated with a better clinical outcome in patients of stratum II. Besides, postimmune sera had the capacity to revert in vitro the immunosuppression induced by NeuGcGM3, as measured by the prevention of CD4 downmodulation on human T lymphocytes. Vaccination had no impact on the frequency of regulatory T cells or circulating NK cells. This study demonstrated, for the first time, the immunogenicity of the NeuGcGM3/VSSP/Montanide ISA 51 vaccine in the adjuvant setting and describes the functionality of induced anti-NeuGcGM3 antibodies as potential surrogate biomarkers of clinical benefit.
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Proteínas de la Membrana Bacteriana Externa/genética , Neoplasias de la Mama/terapia , Vacunas contra el Cáncer/inmunología , Gangliósido G(M3)/análogos & derivados , Neisseria meningitidis/genética , Adyuvantes Inmunológicos/administración & dosificación , Anticuerpos/sangre , Apoptosis , Biomarcadores Farmacológicos/sangre , Neoplasias de la Mama/inmunología , Vacunas contra el Cáncer/genética , Línea Celular Tumoral , Femenino , Gangliósido G(M3)/genética , Humanos , Inmunidad Humoral , Manitol/administración & dosificación , Manitol/análogos & derivados , Persona de Mediana Edad , Neisseria meningitidis/metabolismo , Estadificación de Neoplasias , Ácidos Oléicos/administración & dosificación , Resultado del TratamientoRESUMEN
Vaccine strategies to enhance CD8+ CTL responses remain a current challenge because they should overcome the plasmatic and endosomal membranes for favoring exogenous Ag access to the cytosol of APCs. As a way to avoid this hurdle, sticholysin (St) II, a pore-forming protein from the Caribbean Sea anemone Stichodactyla helianthus, was encapsulated with OVA into liposomes (Lp/OVA/StII) to assess their efficacy to induce a CTL response. OVA-specific CD8+ T cells transferred to mice immunized with Lp/OVA/StII experienced a greater expansion than when the recipients were injected with the vesicles without St, mostly exhibiting a memory phenotype. Consequently, Lp/OVA/StII induced a more potent effector function, as shown by CTLs, in vivo assays. Furthermore, treatment of E.G7-OVA tumor-bearing mice with Lp/OVA/StII significantly reduced tumor growth being more noticeable in the preventive assay. The contribution of CD4+ and CD8+ T cells to CTL and antitumor activity, respectively, was elucidated. Interestingly, the irreversibly inactive variant of the StI mutant StI W111C, encapsulated with OVA into Lp, elicited a similar OVA-specific CTL response to that observed with Lp/OVA/StII or vesicles encapsulating recombinant StI or the reversibly inactive StI W111C dimer. These findings suggest the relative independence between StII pore-forming activity and its immunomodulatory properties. In addition, StII-induced in vitro maturation of dendritic cells might be supporting these properties. These results are the first evidence, to our knowledge, that StII, a pore-forming protein from a marine eukaryotic organism, encapsulated into Lp functions as an adjuvant to induce a robust specific CTL response.
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Adyuvantes Inmunológicos/administración & dosificación , Vacunas contra el Cáncer/inmunología , Venenos de Cnidarios/administración & dosificación , Neoplasias Experimentales/patología , Linfocitos T Citotóxicos/efectos de los fármacos , Animales , Venenos de Cnidarios/inmunología , Femenino , Citometría de Flujo , Liposomas/inmunología , Ratones , Ratones Endogámicos C57BL , Linfocitos T Citotóxicos/inmunologíaRESUMEN
NGcGM3 ganglioside is a tumor-specific antigen expressed in human breast tumors. The NGcGM3/VSSP vaccine, consisting in very small-sized proteoliposomes (VSSP) obtained by the incorporation of NGcGM3 into the outer membrane protein complex of Neisseria meningitidis, has been previously tested in a Phase II trial in patients with metastatic breast cancer (MBC) but emulsified with Montanide ISA 51. An Expanded Access study was carried out in MBC patients aiming to find if a nonemulsive formulation of NGcGM3/VSSP, without Montanide ISA 51, could be more safe and effective. A total of 104 patients were vaccinated with the nonemulsive formulation (900 µg), subcutaneously (SC), or with the emulsive formulation (200 µg), intramuscularly (IM). An intent-to-treat analysis of efficacy was performed with all patients, and 93 patients were split off according to the site of metastases (visceral/nonvisceral). Of note, SC-treated patients exhibited a superior median overall survival (OS) than IM-treated patients (23.6 vs. 8.2 months; log rank P = 0.001). Even though in the subset of patients with nonvisceral metastases SC vaccination duplicated the median OS compared to the alternative option (31.6 vs. 16.5 months), this difference did not reach statistical significance (log rank P = 0.118). Curiously, in patients with visceral metastases, the advantage of the nonemulsive formulation was more apparent (median OS 21.0 vs. 6.2 months; log rank P = 0.005). The vaccine was safe for both formulations.
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Adjuvants are a critical but largely overlooked and poorly understood component included in vaccine formulations to stimulate and modulate the desired immune responses to an antigen. However, unlike in the protective infectious disease vaccines, adjuvants for cancer vaccines also need to overcome the effect of tumor-induced suppressive immune populations circulating in tumor-bearing individuals. Myeloid-derived suppressor cells (MDSC) are considered to be one of the key immunosuppressive populations that inhibit tumor-specific T cell responses in cancer patients. This review focuses on the different signals for the activation of the immune system induced by adjuvants, and the close relationship to the mechanisms of recruitment and activation of MDSC. This work explores the possibility that a cancer vaccine adjuvant may either strengthen or weaken the effect of tumor-induced MDSC, and the crucial need to address this in present and future cancer vaccines.
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Adyuvantes Inmunológicos/uso terapéutico , Vacunas contra el Cáncer/inmunología , Células Mieloides/inmunología , Neoplasias/inmunología , Diferenciación Celular/inmunología , Citocinas/inmunología , Células Dendríticas/citología , Células Dendríticas/inmunología , Humanos , Péptidos y Proteínas de Señalización Intercelular/inmunología , Activación de Linfocitos/inmunología , Macrófagos/citología , Macrófagos/inmunología , Neoplasias/prevención & control , Neoplasias/terapia , Linfocitos T/inmunología , VacunaciónRESUMEN
BACKGROUND: Myeloid-derived suppressor cells (MDSCs) are among the major obstacles that adjuvants for cancer vaccines have to overcome. These cells cross-present tumor-associated antigens (TAA) to naive T lymphocytes with a tolerogenic outcome. Very Small Size Proteoliposomes (VSSP) is used as adjuvant by four therapeutic cancer vaccines currently in Phase I and II clinical trials. We previously found that VSSP reduces the suppressive function of MDSCs, then activating antigen-specific CTL responses in tumor-bearing (TB) mice, with the consequent reduction of tumor growth. However the mechanistic explanation for the immunomodulatory effect of this adjuvant in TB hosts has not been addressed before. METHODS: TB mice were inoculated with VSSP and MDSCs isolated and characterized by their expression of Arg1 and Nos2 genes by RT-PCR. The effect of VSSP on antigen cross-presentation by MDSCs, regulatory T cells (Tregs) expansion and MDSCs differentiation towards dendritic cells (DCs) was analyzed by FACS. Student's t test or ANOVA and Tukey's tests were used for statistical analyses. RESULTS: After inoculating VSSP into TB mice, a significant reduction of Arg1 and Nos2 gene expression was observed in recovered MDSCs. Concurrently the ability of these cells to induce down-regulation of CD3ζ chain on T cells was lost. Likewise in mice inoculated with the adjuvant lower percentages of Tregs were detected. In vitro, VSSP treatment was enough to differentiate MDSCs into phenotypically mature DCs, eliminating the former suppressive effect. Noteworthy, in vivo administration of VSSP to OVA-expressing (EG.7) TB mice abrogated this model antigen cross-presentation by splenic MDSCs. Similar results were obtained even when OVA antigen was administered into these TB mice formulated in VSSP. On the contrary, immunization with the same protein in polyI:C did not change the percentage of MDSCs expressing SIINFEKL/H-2K(b) complexes, whereas a concomitant injection of VSSP aborted the limitations of polyI:C in this setting. CONCLUSIONS: Altogether, these results indicate that VSSP has the peculiar capacity of inhibiting TAA cross-presentation and certain suppressive mechanisms on MDSCs which in turn, combined with the ability to induce differentiation of these cells into antigen-presenting cells (APCs), sustains this adjuvant as an ideal immunomodulator for cancer immunotherapy.
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Leukopenia is a severe condition resulting from both pathological processes and some treatments, like chemotherapy in cancer patients. However, the activation of the patient immune system is required for the success of immunotherapeutic strategies, as cancer vaccines. In this regard, leukopenia constitutes a major hurdle to overcome, mainly due to the impairment of cytotoxic T lymphocyte (CTL) responses. Adjuvants are basic components of vaccine formulations, which might be useful to stimulate immunity under this immunosuppressed condition. To this aim, we tested the capacity of a novel nanoparticulated complex, very small size proteoliposomes (VSSP), to promote CTL even in a leukopenic scenario. Noteworthy, we observed that a VSSP-based OVA vaccine induced a normal antigen-specific CTL response in mice rendered leukopenia by the administration of high doses of the chemotherapeutic agent cyclophosphamide (CY), while under the same conditions the OVA antigen formulated in the TLR-3 agonist polyinosinic-polycytidylic acid (P(I:C)) was ineffective. Moreover, an appropriate combination of VSSP with the P(I:C) vaccine was able to restore the CD8(+) T cell effector function in leukopenic mice. VSSP induced not only a faster repopulation of immune cells in CY-receiving animals, but also enhanced the recovery of memory T lymphocytes and myeloid dendritic cells (DCs) while simultaneously abrogated the immunosuppressive capacity of myeloid-derived suppressor cells (MDSCs). Our results suggest that VSSP could be a particularly suitable immunomodulator to be used in CTL-promoting active immunotherapy strategies operating in severe immune compromised scenarios.
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Adyuvantes Inmunológicos/administración & dosificación , Leucopenia/inmunología , Neisseria meningitidis/química , Ovalbúmina/inmunología , Proteolípidos/administración & dosificación , Linfocitos T Citotóxicos/inmunología , Adyuvantes Inmunológicos/aislamiento & purificación , Animales , Femenino , Ratones , Ratones Endogámicos C57BL , Ovalbúmina/administración & dosificación , Proteolípidos/aislamiento & purificaciónRESUMEN
The interaction between cancer vaccine adjuvants and myeloid-derived suppressor cells (MDSCs) is currently poorly understood. Very small size proteoliposomes (VSSP) are a nanoparticulated adjuvant under investigation in clinical trials in patients with renal carcinoma, breast cancer, prostate cancer, and cervical intraepithelial neoplasia grade III. We found that VSSP adjuvant induced a significant splenomegaly due to accumulation of CD11b(+)Gr-1(+) cells. However, VSSP-derived MDSCs showed a reduced capacity to suppress both allogeneic and Ag-specific CTL response compared with that of tumor-induced MDSCs. Moreover, splenic MDSCs isolated from tumor-bearing mice treated with VSSP were phenotypically more similar to those isolated from VSSP-treated tumor-free mice and much less suppressive than tumor-induced MDSCs, both in vitro and in vivo. Furthermore, different from dendritic cell vaccination, inoculation of VSSP-based vaccine in EG.7-OVA tumor-bearing mice was sufficient to avoid tumor-induced tolerance and stimulate an immune response against OVA Ag, similar to that observed in tumor-free mice. This effect correlated with an accelerated differentiation of MDSCs into mature APCs that was promoted by VSSP. VSSP used as a cancer vaccine adjuvant might thus improve antitumor efficacy not only by stimulating a potent immune response against tumor Ags but also by reducing tumor-induced immunosuppression.
Asunto(s)
Adyuvantes Inmunológicos/administración & dosificación , Antineoplásicos/administración & dosificación , Vacunas contra el Cáncer/administración & dosificación , Linfoma/prevención & control , Células Mieloides/inmunología , Nanopartículas/administración & dosificación , Proteolípidos/administración & dosificación , Sarcoma Experimental/prevención & control , Animales , Proteínas de la Membrana Bacteriana Externa/administración & dosificación , Proteínas de la Membrana Bacteriana Externa/inmunología , Vacunas contra el Cáncer/inmunología , Línea Celular Tumoral , Femenino , Gangliósido G(M3)/administración & dosificación , Gangliósido G(M3)/inmunología , Inhibidores de Crecimiento/administración & dosificación , Linfoma/inmunología , Linfoma/patología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Transgénicos , Células Mieloides/patología , Neisseria meningitidis/inmunología , Proteolípidos/inmunología , Sarcoma Experimental/inmunología , Sarcoma Experimental/patologíaRESUMEN
Nimotuzumab is an EGFR-targeting antibody that has demonstrated encouraging clinical results in the absence of severe side-effects observed with other approved anti-EGFR antibodies. We investigated whether different clinical behavior of nimotuzumab is related to its bivalent/monovalent binding profile. Binding properties of nimotuzumab and cetuximab, the most development of anti-EGFR antibodies, were studied in vitro using chip surfaces and cells with varying EGFR expression levels. Experimental observations demonstrated that in contrast to cetuximab, the intrinsic properties of nimotuzumab required bivalent binding for stable attachment to the cellular surface, leading to nimotuzumab selectively binding to cells that express moderate to high EGFR expression levels. At these conditions, both antibodies bound bivalently, and accumulated to similar degrees. When EGFR density is low, nimotuzumab monovalent interaction was transient, whereas cetuximab continued to interact strongly with the receptors. We compared the in vitro anti-tumor efficacy of nimotuzumab and cetuximab. Cetuximab decreased the cell viability and induced apoptosis for all the tested cell lines, effects which did not depend on EGFR expression level. In contrast, nimotuzumab also provoked significant anti-cellular effects, but its anti-tumor capacity decreased together with EGFR expression level. Cetuximab Fab fragment was able to impact tumor cell survival, whereas nimotuzumab fragment totally lost this effect. Tumor-xenograft experiments using cells with a high EGFR expression revealed similar tumor growth inhibiting effects for both antibodies. This study suggests an explanation for nimotuzumab clinical profile, whereby anti-tumor activity is obtained in absence of severe toxicities due to its properties of bivalent binding to EGFR.