Post-translational Modification of the NKG2D Ligand RAET1G Leads to Cell Surface Expression of a Glycosylphosphatidylinositol-linked Isoform

NKG2D is an important activating receptor on lymphocytes. In human, it interacts with two groups of ligands: the major histocompatibility complex class I chain-related A/B (MICA/B) family and the UL-16 binding protein (ULBP) family, also known as retinoic acid early transcript (RAET1). MIC proteins are membrane-anchored, but all of the ULBP/RAET1 proteins, except for RAET1E and RAET1G, are glycosylphosphatidylinositol (GPI)-anchored. To address the reason for these differences we studied the association of RAET1G with the membrane. Using epitope-tagged RAET1G protein in conjunction with antibodies to different parts of the molecule and in pulse-chase experiments, we showed that the C terminus of the protein was cleaved soon after protein synthesis. Endoglycosidase H and peptide N-glycosidase treatment and cell surface immunoprecipitation indicated that most of the protein stayed in the endoplasmic reticulum, but some of the cleaved form was modified in the Golgi and transported to the cell surface. We examined the possibility of GPI anchoring of the protein in three ways: (i) Phosphatidylinositol (PI)-specific phospholipase C released the PI-linked form of the protein. (ii) The surface expression pattern of RAET1G decreased in cells defective in GPI anchoring through mutant GPI-amidase. (iii) Site-directed mutagenesis, to disrupt residues predicted to facilitate GPI-anchoring, resulted in diminished surface expression of RAET1G. Thus, a form of RAET1G is GPI-anchored, in line with most other ULBP/RAET1 family proteins. The cytoplasmic tail and transmembrane domains appear to result from gene duplication and frameshift mutation. Together with our previous results, our data suggest that RAET1G is regulated post-translationally to produce a GPI-anchored isoform

First results of the Am-241(n,f) cross section measurement at the Experimental Area 2 of the n_TOF facility at CERN

This research is co-financed by Greece and the European Union (European Social Fund-ESF) through the Operational Programme Human Resources Development, Education and Lifelong Learning in the context of the project "Strengthening Human Resources Research Potential via Doctorate Research" (MIS-5000432), implemented by the State Scholarships Foundation (IKY). Also, the authors would like to acknowledge the support of the European Commission under the CHANDA project (7th Framework Programme).Feasibility, design and sensitivity studies on innovative nuclear reactors that could address the issue of nuclear waste transmutation using fuels enriched in minor actinides, require high accuracy cross section data for a variety of neutron-induced reactions from thermal energies to several tens of MeV. The isotope Am-241 (T-1/2= 433 years) is present in high-level nuclear waste (HLW), representing about 1.8 % of the actinide mass in spent PWR UOx fuel. Its importance increases with cooling time due to additional production from the beta-decay of Pu-241 with a half-life of 14.3 years. The production rate of 241Am in conventional reactors, including its further accumulation through the decay of Pu-241 and its destruction through transmutation/incineration are very important parameters for the design of any recycling solution. In the present work, the Am-241(n,f) reaction cross-section was measured using Micromegas detectors at the Experimental Area 2 of the n_TOF facility at CERN. For the measurement, the U-235(n,f) and U-238(n,f) reference reactions were used for the determination of the neutron flux. In the present work an overview of the experimental setup and the adopted data analysis techniques is given along with preliminary results.European Union (European Social Fund-ESF) through the Operational Programme Human Resources Development, Education and Lifelong Learning MIS-5000432European Commission under the CHANDA project (7th Framework Programme

Intercellular transfer and supramolecular organization of human leukocyte antigen C at inhibitory natural killer cell immune synapses

17.10.14 KB. OK to add published version to Spiral, RUP polic

Measurement of the neutron-induced fission cross section of Th 230 at the CERN n_TOF facility

The neutron-induced fission cross section of 230 Th has been measured at the neutron time-of-flight facility n_TOF located at CERN. The experiment was performed at the experimental area EAR-1 with a neutron flight path of 185 m, using Micromegas detectors for the detection of the fission fragments. The 230 Th(n, f ) cross section was determined relative to the 235 U(n, f ) one, covering the energy range from the fission threshold up to 400 MeV. The results from the present work are compared with existing cross-section datasets and the observed discrepancies are discussed and analyzed. Finally, using the code EMPIRE 3.2.3 a theoretical study, based on the statistical model, was performed leading to a satisfactory reproduction of the experimental results with the proper tuning of the respective parameters, while for incident neutron energy beyond 200 MeV the fission of 230 Th was described by Monte Carlo simulations.This project received funding from the Euratom “Support safe operation of nuclear systems” program 2014–2018 under Grant Agreement No. 847552 (SANDA) and by the funding agencies of the participating institutes. This research is imple- mented through the IKY scholarships program and cofinanced by the European Union (European Social Fund ’ESF) and Greek national funds through the action entitled “Reinforce- ment of Postdoctoral Researchers - 2nd call (MIS 5033021)”, in the framework of the Operational Programme “Human Resources Development Program, Education and Lifelong Learning” of the National Strategic Reference Framework.Article signat per 137 autors/es: V. Michalopoulou, A. Stamatopoulos, M. Diakaki, A. Tsinganis, R. Vlastou, M. Kokkoris, N. Patronis, Z. Eleme, D. Macina, L. Tassan-Got, N. Colonna, E. Chiaveri, A. Ventura, P. Schillebeeckx, J. Heyse, G. Sibbens, G. Alaerts, A. Borella, A. Moens, D. Vanleeuw, O. Aberle, V. Alcayne, S. Amaducci, J. Andrzejewski, L. Audouin, V. Babiano-Suarez, M. Bacak, M. Barbagallo, S. Bennett, E. Berthoumieux, J. Billowes, D. Bosnar, A. Brown, M. Busso, M. Caamaño, L. Caballero, F. Calviño, M. Calviani, D. Cano-Ott, A. Casanovas, F. Cerutti, G. Cortés, M. A. Cortés-Giraldo, L. Cosentino, S. Cristallo, L. A. Damone, P. J. Davies, M. Dietz, C. Domingo-Pardo, R. Dressler, Q. Ducasse, E. Dupont, I. Durán, B. Fernández-Domínguez, A. Ferrari, P. Finocchiaro, V. Furman, K. Göbel, R. Garg, A. Gawlik-Ramiega, S. Gilardoni, I. F. Gonçalves, E. González-Romero, C. Guerrero, F. Gunsing, H. Harada, S. Heinitz, D. G. Jenkins, A. Junghans, F. Käppeler, Y. Kadi, A. Kimura, I. Knapová, Y. Kopatch, M. Krticka, D. Kurtulgil, I. Ladarescu, C. Lederer-Woods, H. Leeb, J. Lerendegui-Marco, S. J. Lonsdale, A. Manna, T. Martínez, A. Masi, C. Massimi, P. Mastinu, M. Mastromarco, E. A. Maugeri, A. Mazzone, E. Mendoza, A. Mengoni, P. M. Milazzo, F. Mingrone, J. Moreno-Soto, A. Musumarra, A. Negret, R. Nolte, F. Ogállar, A. Oprea, A. Pavlik, J. Perkowski, C. Petrone, L. Piersanti, E. Pirovano, I. Porras, J. Praena, J. M. Quesada, D. Ramos-Doval, T. Rauscher, R. Reifarth, D. Rochman, Y. Romanets, C. Rubbia, M. Sabaté-Gilarte, A. Saxena, D. Schumann, A. Sekhar, A. G. Smith, N. V. Sosnin, P. Sprung, G. Tagliente, J. L. Tain, A. Tarifeño-Saldivia, Th. Thomas, P. Torres-Sánchez, J. Ulrich, S. Urlass, S. Valenta, G. Vannini, V. Variale, P. Vaz, D. Vescovi, V. Vlachoudis, A. Wallner, P. J. Woods, T. Wright, and P. Žugec.Postprint (published version

First results of the 241Am(n,f) cross section measurement at the Experimental Area 2 of the n_TOF facility at CERN

Feasibility, design and sensitivity studies on innovative nuclear reactors that could address the issue of nuclear waste transmutation using fuels enriched in minor actinides, require high accuracy cross section data for a variety of neutron-induced reactions from thermal energies to several tens of MeV. The isotope 241Am (T1/2= 433 years) is present in high-level nuclear waste (HLW), representing about 1.8 % of the actinide mass in spent PWR UOx fuel. Its importance increases with cooling time due to additional production from the β-decay of 241Pu with a half-life of 14.3 years. The production rate of 241 Am in conventional reactors, including its further accumulation through the decay of 241Pu and its destruction through transmutation/incineration are very important parameters for the design of any recycling solution. In the present work, the 241 Am(n,f) reaction cross-section was measured using Micromegas detectors at the Experimental Area 2 of the n_TOF facility at CERN. For the measurement, the 235U(n,f) and 238U(n,f) reference reactions were used for the determination of the neutron flux. In the present work an overview of the experimental setup and the adopted data analysis techniques is given along with preliminary results

Cell surface organization of stress-inducible proteins ULBP and MICA that stimulate human NK cells and T cells via NKG2D

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NEAR at n_TOF/CERN: Preparing the first multi-foil activation measurement

The n_TOF facility at CERN is a neutron Time-Of-Flight facility based on a spallation neutron source. During the Second Long Shutdown (LS2) , a new experimental zone was designed and delivered. This new experimental area -the NEAR station - is located very close to the lead spallation target, at a distance of just ~3m. In this way, the high luminosity of the n_TOF neutron spallation source can be fully exploited. Towards the characterization of the new experimental area as well for the benchmarking of the performed simulations, the multi-foil activation measurement will be implemented. Eleven threshold and seven capture reference reactions will be utilized for the unfolding of the NEAR neutron beam energy spectrum that stretches from the meV to the GeV region

Human Ovarian Tumor Cells Escape γδ T Cell Recognition Partly by Down Regulating Surface Expression of MICA and Limiting Cell Cycle Related Molecules

Background: Mechanisms of human Vc2Vd2 T cell-mediated tumor immunity have yet to be fully elucidated. Methods and Findings: At least some tumor cell recognition is mediated by NKG2D-MICA interactions. Herein, by using MTT assay and PI-BrdU co-staining and Western-blot, we show that these Vc2Vd2 T cells can limit the proliferation of ovarian tumor cells by down regulation of apoptosis and cell cycle related molecules in tumor cells. Cell-to-cell contact is critical. cd T cell-resistant, but not susceptible ovarian tumor cells escape cd T cell-mediated immune recognition by up-regulating pErk1/2, thereby decreasing surface MICA levels. Erk1/2 inhibitor pretreatment or incubation prevents this MICA decrease, while up-regulating key cell cycle related molecules such as CDK2, CDK4 and Cyclin D1, as well as apoptosis related molecules making resistant tumor cells now vulnerable to cd T cell-mediated lysis. Conclusion: These findings demonstrate novel effects of cdT cells on ovarian tumor cells

Measurement of the Yb 176 (n,γ) cross section at the n_TOF facility at CERN

© 2024 authors. Published by the American Physical Society. Published by the American Physical Society under the terms of the "https://creativecommons.org/licenses/by/4.0/"Creative Commons Attribution 4.0 International license.Background: The Yb176(n,γ)Yb177→Lu177 reaction is of interest in nuclear medicine as it is the preferred production route for Lu177. This radioisotope has seen a very fast growth of usage in nuclear medicine in recent years due to its outstanding properties. New data on this reaction could provide useful information for production at new facilities. Purpose: We aim to resolve resonances in the Yb176(n,γ)Yb177 reaction for the first time. Previous capture measurement provided data at thermal point and encompassed integral measurements in the range from 3 keV to 1 MeV, where three time-of-flight measurements are available, but with low resolution to resolve the resonances. Transmission measurements from the 1970s resolved and analyzed some resonances. Method: We measure the neutron capture cross section of Yb176(n,γ)Yb177 by means of the time-of-flight technique at the Experimental Area 1 of the n_TOF facility at CERN using an enriched Yb1762O3 sample and an array of four C6D6 liquid scintillation detectors. Results: We have resolved 164 resonances up to 21 keV, including 96 new ones. We also provide new capture experimental data from 90 eV to 3 keV, and we extend the resolved resonance region up to 21 keV. In addition, resonance decay widths, Γγ and Γn, are provided for all resonances together with resonance energies. Conclusions: The Yb176(n,γ)Yb177 reaction has been measured, providing resonance parameters for the first time from a few eV to 21 keV. The analysis of the resonances has been carried out and compared with previous works and existing libraries, revealing discrepancies due to the new information on Γγ parameters. Our results are consistent with the Γn parameters obtained in transmission measurements.Peer reviewe