1,734 research outputs found
Adoptive immunotherapy with Cl-IB-MECA-treated CD8+T cells reduces melanoma growth in mice
Cl-IB-MECA is a selective A3 adenosine receptor agonist, which plays a crucial role in limiting tumor progression. In mice, Cl-IB-MECA administration enhances the anti-tumor T cell-mediated response. However, little is known about the activity of Cl-IB-MECA on CD8+ T cells. The aim of this study was to investigate the effect of ex vivo Cl-IB-MECA treatment of CD8+ T cells, adoptively transferred in melanoma-bearing mice. Adoptive transfer of Cl-IB-MECA-treated CD8+ T cells or a single administration of Cl-IB-MECA (20 ng/mouse) inhibited tumor growth compared with the control group and significantly improved mouse survival. This was associated with the release of Th1-type cytokines and a greater influx of mature Langerin+ dendritic cells (LCs) into the tumor microenvironment. CD8+ T cells treated with Cl-IB-MECA released TNF-α which plays a critical role in the therapeutic efficacy of these cells when injected to mice. Indeed, neutralization of TNF-α by a specific monoclonal Ab significantly blocked the anti-tumor activity of Cl-IB-MECA-treated T cells. This was due to the reduction in levels of cytotoxic cytokines and the presence of fewer LCs. In conclusion, these studies reveal that ex vivo treatment with Cl-IB-MECA improves CD8+ T cell adoptive immunotherapy for melanoma in a TNF-α-dependent manner
Robust gap repair in the contractile ring ensures timely completion of cytokinesis.
Cytokinesis in animal cells requires the constriction of an actomyosin contractile ring, whose architecture and mechanism remain poorly understood. We use laser microsurgery to explore the biophysical properties of constricting rings in Caenorhabditis elegans embryos. Laser cutting causes rings to snap open. However, instead of disintegrating, ring topology recovers and constriction proceeds. In response to severing, a finite gap forms and is repaired by recruitment of new material in an actin polymerization-dependent manner. An open ring is able to constrict, and rings repair from successive cuts. After gap repair, an increase in constriction velocity allows cytokinesis to complete at the same time as controls. Our analysis demonstrates that tension in the ring increases while net cortical tension at the site of ingression decreases throughout constriction and suggests that cytokinesis is accomplished by contractile modules that assemble and contract autonomously, enabling local repair of the actomyosin network. Consequently, cytokinesis is a highly robust process impervious to discontinuities in contractile ring structure.This project has received funding from the European Research Council
(grants 640553, 260892, and 338410), Fundo Europeu de
Desenvolvimento Regional (FED ER) funds through the Operational
Competitiveness Program (COM PETE), national funds through
Fundação para a Ciência e a Tecnologia (FCT) under the project
FCO MP-01-0124-FED ER-028255 (PTDC/BEX-BCM/0654/2012),
Fundação Luso-Americana para o Desenvolvimento Life Science
2020, and the Louis-Jeantet Young Investigator Award to H. Maiato.
A.X. Carvalho, R. Gassmann, and I.A. Telley have FCT Investigator
positions funded by FCT and cofunded by the European Social Fund
through Programa Operacional Temático Potencial Type 4.2 promotion
of scientific employment. A.M. Silva holds an FCT fellowship
(SFRH/BPD/95707/2013). D.S. Osório was cofunded by the Programa
Operacional Regional do Norte under the Quadro de
Downloaded from jcb.rupress.org on February 27, 2018
Laser microsurgery in the contractile ring • Silva et al. 799
Referência Estratégico Nacional through FED ER and by FCT grant
NOR TE-07-0124-FED ER-000003 (Cell Homeostasis Tissue Organization
and Organism Biology)
Inhibition of CD73 improves B cell-mediated anti-tumor immunity in a mouse model of melanoma.
CD73 is a cell surface enzyme that suppresses T cell-mediated immune responses by producing extracellular adenosine. Growing evidence suggests that targeting CD73 in cancer may be useful for an effective therapeutic outcome. In this study, we demonstrate that administration of a specific CD73 inhibitor, adenosine 5'-(α,β-methylene)diphosphate (APCP), to melanoma-bearing mice induced a significant tumor regression by promoting the release of Th1- and Th17-associated cytokines in the tumor microenvironment. CD8+ T cells were increased in melanoma tissue of APCP-treated mice. Accordingly, in nude mice APCP failed to reduce tumor growth. Importantly, we observed that after APCP administration, the presence of B cells in the melanoma tissue was greater than that observed in control mice. This was associated with production of IgG2b within the melanoma. Depletion of CD20+ B cells partially blocked the anti-tumor effect of APCP and significantly reduced the production of IgG2b induced by APCP, implying a critical role for B cells in the anti-tumor activity of APCP. Our results also suggest that APCP could influence B cell activity to produce IgG through IL-17A, which significantly increased in the tumor tissue of APCP-treated mice. In support of this, we found that in melanoma-bearing mice receiving anti-IL-17A mAb, the anti-tumor effect of APCP was ablated. This correlated with a reduced capacity of APCP-treated mice to mount an effective immune response against melanoma, as neutralization of this cytokine significantly affected both the CD8+ T cell- and B cell-mediated responses. In conclusion, we demonstrate that both T cells and B cells play a pivotal role in the APCP-induced anti-tumor immune response
Biologically relevant effects of mRNA amplification on gene expression profiles
BACKGROUND: Gene expression microarray technology permits the analysis of global gene expression profiles. The amount of sample needed limits the use of small excision biopsies and/or needle biopsies from human or animal tissues. Linear amplification techniques have been developed to increase the amount of sample derived cDNA. These amplified samples can be hybridised on microarrays. However, little information is available whether microarrays based on amplified and unamplified material yield comparable results. In the present study we compared microarray data obtained from amplified mRNA derived from biopsies of rat cardiac left ventricle and non-amplified mRNA derived from the same organ. Biopsies were linearly amplified to acquire enough material for a microarray experiment. Both amplified and unamplified samples were hybridized to the Rat Expression Set 230 Array of Affymetrix. RESULTS: Analysis of the microarray data showed that unamplified material of two different left ventricles had 99.6% identical gene expression. Gene expression patterns of two biopsies obtained from the same parental organ were 96.3% identical. Similarly, gene expression pattern of two biopsies from dissimilar organs were 92.8% identical to each other. Twenty-one percent of reporters called present in parental left ventricular tissue disappeared after amplification in the biopsies. Those reporters were predominantly seen in the low intensity range. Sequence analysis showed that reporters that disappeared after amplification had a GC-content of 53.7+/-4.0%, while reporters called present in biopsy- and whole LV-samples had an average GC content of 47.8+/-5.5% (P <0.001). Those reporters were also predicted to form significantly more (0.76+/-0.07 versus 0.38+/-0.1) and longer (9.4+/-0.3 versus 8.4+/-0.4) hairpins as compared to representative control reporters present before and after amplification. CONCLUSION: This study establishes that the gene expression profile obtained after amplification of mRNA of left ventricular biopsies is representative for the whole left ventricle of the rat heart. However, specific gene transcripts present in parental tissues were undetectable in the minute left ventricular biopsies. Transcripts that were lost due to the amplification process were not randomly distributed, but had higher GC-content and hairpins in the sequence and were mainly found in the lower intensity range which includes many transcription factors from specific signalling pathways
E/S0 Galaxies on the Blue Color-Stellar Mass Sequence at z=0: Fading Mergers or Future Spirals?
We identify a population of morphologically defined E/S0 galaxies lying on
the blue sequence at the present epoch. Using three samples, we analyze
blue-sequence E/S0s with stellar masses >10^8 Msun, arguing that individual
objects may be evolving either up toward the red sequence or down into the blue
sequence. Blue-sequence E/S0 galaxies become more common with decreasing
stellar mass, comprising <2% of E/S0s near the "shutdown mass" M_s ~ 1-2 x
10^11 Msun, increasing to >5% near the "bimodality mass" M_b ~ 3 x 10^10 Msun,
and sharply rising to >20-30% below the "threshold mass" M_t ~ 4-6 x 10^9 Msun.
The strong emergence of blue-sequence E/S0s below M_t coincides with a
previously reported global increase in mean atomic gas fractions below M_t for
galaxies of all types on both sequences, suggesting that the availability of
cold gas may be basic to blue-sequence E/S0s' existence. Environmental analysis
reveals that many sub-M_b blue-sequence E/S0s reside in low to intermediate
density environments. In mass-radius and mass-sigma scaling relations,
blue-sequence E/S0s are more similar to red-sequence E/S0s than to late-type
galaxies, but they represent a transitional class. While some of them,
especially in the high-mass range from M_b to M_s, resemble major-merger
remnants that will likely fade onto the red sequence, most blue-sequence E/S0s
below M_b show signs of disk and/or pseudobulge building, which may be enhanced
by companion interactions. We argue that sub-M_b blue-sequence E/S0s occupy a
"sweet spot" in stellar mass and concentration, with both abundant gas and
optimally efficient star formation, which may enable the formation of large
spiral disks. [abridged]Comment: AJ, submitted, revised, 21 pages with 15 figures (one in two parts,
one color); full resolution version available at
http://www.physics.unc.edu/~sheila/kgb.pd
Copy number variants prioritization after array-CGH analysis - a cohort of 1000 patients
Array-based comparative genomic hybridization has been assumed to be the first genetic test offered to detect genomic imbalances in patients with unexplained intellectual disability with or without dysmorphisms, multiple congenital anomalies, learning difficulties and autism spectrum disorders. Our study contributes to the genotype/phenotype correlation with the delineation of laboratory criteria which help to classify the different copy number variants (CNVs) detected. We clustered our findings into five classes ranging from an imbalance detected in a microdeletion/duplication syndrome region (class I) to imbalances that had previously been reported in normal subjects in the Database of Genomic Variants (DGV) and thus considered common variants (class IV).info:eu-repo/semantics/publishedVersio
The effects of symmetry on the dynamics of antigenic variation
In the studies of dynamics of pathogens and their interactions with a host
immune system, an important role is played by the structure of antigenic
variants associated with a pathogen. Using the example of a model of antigenic
variation in malaria, we show how many of the observed dynamical regimes can be
explained in terms of the symmetry of interactions between different antigenic
variants. The results of this analysis are quite generic, and have wider
implications for understanding the dynamics of immune escape of other
parasites, as well as for the dynamics of multi-strain diseases.Comment: 21 pages, 4 figures; J. Math. Biol. (2012), Online Firs
Hydrolysis optimization and characterization study of preparing fatty acids from Jatropha curcas seed oil
<p>Abstract</p> <p>Background</p> <p>Fatty acids (FAs) are important as raw materials for the biotechnology industry. Existing methods of FAs production are based on chemical methods. In this study potassium hydroxide (KOH)-catalyzed reactions were utilized to hydrolysis <it>Jatropha curcas </it>seed oil.</p> <p>Results</p> <p>The parameters effect of ethanolic KOH concentration, reaction temperature, and reaction time to free fatty acid (FFA%) were investigated using D-Optimal Design. Characterization of the product has been studied using Fourier transforms infrared spectroscopy (FTIR), gas chromatography (GC) and high performance liquid chromatography (HPLC). The optimum conditions for maximum FFA% were achieved at 1.75M of ethanolic KOH concentration, 65°C of reaction temperature and 2.0 h of reaction time.</p> <p>Conclusions</p> <p>This study showed that ethanolic KOH concentration was significant variable for <it>J. curcas </it>seed oil hydrolysis. In a 18-point experimental design, FFA% of hydrolyzed <it>J. curcas </it>seed oil can be raised from 1.89% to 102.2%, which proved by FTIR and HPLC.</p
Surface modification of a biodegradable composite by UV laser ablation : in vitro biological performance
Melt blends of chitosan and biodegradable aliphatic polyester have been physically and biologically
studied, presenting great potential for biomedical applications. Structurally, poly(butylene
succinate)–chitosan (PBS/Cht) composite scaffolds are covered by a thin PBS layer, preventing
the desired interaction of cells/tissues with the chitosan particules. In the present work, a
selective and controlled ablation of this skin layer was induced by UV laser processing. X-ray
photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF–SIMS)
data demonstrated an increment of chitosan components and others resulting from the laser
ablation process. The biological activity (i.e. cell viability and proliferation) on the inner regions of
the composite scaffolds is not significantly different from those of the external layer, despite the
observed differences in surface roughness (determined by interferometric optical profilometry) and
wettability (water contact angle). However, the morphology of human osteoblastic cells was found
to be considerably different in the case of laser-processed samples, since the cells tend to aggregate
in multilayer columnar structures, preferring the PBS surface and avoiding the chitosan-rich areas.
Thus, UV laser ablation can be considered a model technique for the physical surface modification
of biomaterials without detrimental effects on cellular activity.This work was partially supported by the European Union Integrated Project GENOSTEM (LSH-STREP-CT-2003-503161), the European Union Network of Excellence EXPERTISSUES (NMP3-CT-2004-500283), the Interreg III Project PROTEUS (SP1P151/03) and Xunta de Galicia (Consolidacion 2006/12). The Portuguese Foundation for Science and Technology is also acknowledged for a PhD grant to A.M. (SFRH/BD/24382/2005). The authors wish to thank C. Serra from CACTI of the University of Vigo for the XPS and ToF-SIMS measurements
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