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International audience; IL‐7 is an important cytokine for T cell lymphopoiesis. Blockade of the IL‐7 signal‐ing pathway has been shown to induce long‐term graft survival or graft tolerance in murine transplant models through inhibiting T cell homeostasis and favoring im ‐munoregulation. In this study, we assessed for the first time the effects of a blocking anti‐human cluster of differentiation 127 (CD127) mAb administered in combination with low‐dose tacrolimus or thymoglobulin in a life‐sustaining kidney allograft model in baboons. Contrary to our expectation, the addition of an anti‐CD127 mAb to the treatment protocols did not prolong graft survival compared to low‐dose tacrolimus alone or thymoglobulin alone. Anti‐CD127 mAb administration led to full CD127 re ‐ceptor occupancy during the follow‐up period. However, all treated animals lost their kidney graft between 1 week and 2 weeks after transplantation. Unlike in rodents, in nonhuman primates, anti‐CD127 mAb treatment does not decrease the absolute numbers of lymphocyte and lymphocyte subsets and does not effectively inhibit postdepletional T cell proliferation and homeostasis, suggesting that IL‐7 is not a lim ‐iting factor for T cell homeostasis in primates.

We present a new 3D magnetohydrodynamic (MHD) heliospheric model for space-weather forecasting driven by boundary conditions defined from white-light observations of the solar corona. The model is based on the MHD code PLUTO, constrained by an empirical derivation of the solar wind background properties at 0.1 au. This empirical method uses white-light observations to estimate the position of the heliospheric current sheet (HCS). The boundary conditions necessary to run HelioCast are then defined from pre-defined relations between the necessary MHD properties (speed, density, and temperature) and the distance to the current sheet. We assess the accuracy of the model over six Carrington rotations during the first semester of 2018. Using point-by-point metrics and event-based analyses, we evaluate the performances of our model varying the angular width of the slow solar wind layer surrounding the HCS. We also compare our empirical technique with two well-tested models of the corona: Multi-VP and WindPredict-AW. We find that our method is well-suited to reproduce high-speed streams, and does – for well-chosen parameters – better than full MHD models. The model shows, nonetheless, limitations that could worsen for rising and maximum solar activity.

International audience

In-situ measurements by several spacecraft have revealed that the solar wind is frequently perturbed by transient structures (magnetic folds, jets, waves, flux-ropes) that propagate rapidly away from the Sun over large distances. Parker Solar Probe has detected frequent rotations of the magnetic field vector at small heliocentric distances, accompanied by surprisingly large solar wind rotation rates. The physical origin of such magnetic field bends, the conditions for their survival across the interplanetary space, and their relation to solar wind rotation are yet to be clearly understood. We traced measured solar wind flows from the spacecraft position down to the surface of the Sun to identify their potential source regions and used a global MHD model of the corona and solar wind to relate them to the rotational state of the low solar corona. We identified regions of the solar corona for which solar wind speed and rotational shear are important and long-lived, that can be favourable to the development of magnetic deflections and to their propagation across extended heights in the solar wind. We show that coronal rotation is highly structured and that enhanced flow shear develops near the boundaries between coronal holes and streamers, around and above pseudo-streamers, even when such boundaries are aligned with the direction of solar rotation. A large fraction of the switchbacks identified by PSP map back to these regions, both in terms of instantaneous magnetic field connectivity and of the trajectories of wind streams that reach the spacecraft. These regions of strong shears are likely to leave an imprint on the solar wind over large distances and to increase the transverse speed variability in the slow solar wind. The simulations and connectivity analysis suggest they can be a source of the switchbacks and spikes observed by Parker Solar Probe., Comment: Accepted for publication in Astronomy & Astrophysics

The BAG3- and SIRPα- mediated pathways trigger distinct cellular targets and signaling mechanisms in pancreatic cancer microenvironment. To explore their functional connection, we investigated the effects of their combined blockade on cancer growth in orthotopic allografts of pancreatic cancer mt4–2D cells in immunocompetent mice. The anti-BAG3 + anti-SIRPα mAbs treatment inhibited (p = 0.007) tumor growth by about the 70%; also the number of metastatic lesions was decreased, mostly by the effect of the anti-BAG3 mAb. Fibrosis and the expression of the CAF activation marker α-SMA were reduced by about the 30% in animals treated with anti-BAG3 mAb compared to untreated animals, and appeared unaffected by treatment with the anti-SIRPα mAb alone; however, the addition of anti-SIRPα to anti-BAG3 mAb in the combined treatment resulted in a > 60% (p p p

International audience; Myeloid-derived suppressor cells (MDSC) are a heterogeneous population of immature hematopoietic precursors known to suppress immune responses. Interaction of SIRP alpha (SIRPα), expressed by myeloid cells, with the ubiquitous receptor CD47 is an important immune checkpoint of the innate response regulating macrophages and dendritic cells functions. We previously described that MDSC expressing SIRPα accumulated after transplantation and maintained kidney allograft tolerance. However, the role of the SIRPα/CD47 axis on MDSC function remained unknown. Here, we found that blocking SIRPα or CD47 with monoclonal antibodies (mAbs) induced differentiation of MDSC into myeloid cells overexpressing MHC class II, CD86 costimulatory molecule and increased secretion of macrophage-recruiting chemokines (eg, MCP-1). Using a model of long-term kidney allograft tolerance sustained by MDSC, we observed that administration of blocking anti-SIRPα or CD47 mAbs induced graft dysfunction and rejection. Loss of tolerance came along with significant decrease of MDSC and increase in MCP-1 concentration in the periphery. Graft histological and transcriptomic analyses revealed an inflammatory (M1) macrophagic signature at rejection associated with overexpression of MCP-1 mRNA and protein in the graft. These findings indicate that the SIRPα-CD47 axis regulates the immature phenotype and chemokine secretion of MDSC and contributes to the induction and the active maintenance of peripheral acquired immune tolerance.

Two distinct plasma states of slow solar wind are identified from in-situ measurements of the local magnetic field and the solar wind plasma by Parker Solar Probe (PSP) during Encounter 2. So close to the Sun the wind measured at PSP has not yet undergone too many transformations related to the expansion and propagation of the wind in the heliosphere and we focus on the wind intervals with no magnetic switchbacks. The two states differ by plasma beta, mass flux and magnetic pressure. A detailed study of the spacecraft connectivity, combined with the optimised PFSS-reconstructed solar magnetic field (from magnetic field maps and EUV observations of coronal holes), revealed that the boundary between the two states can be interpreted as a transition from a streamer-like to an equatorial coronal hole plasma flow. The streamer type tube has an expansion factor 20 times higher than the equatorial coronal hole tube, and a stronger expansion over a larger spatial extent. The simulations reproduce the properties observed in the data: slower and denser wind at the edge of the streamer compared to equatorial coronal hole plasma. Temperature profiles from simulations provide an explanation: due to the difference of photospheric magnetic field and different flux tube geometries, heating is more intense and lower along a streamer-like tube than along a coronal hole-like tube, even if footpoints of both tubes are very close to each other on the solar surface. This results in plasma outflows with different background properties at the edge of the streamer and at the boundary of the equatorial coronal hole, independently of the solar wind variability., The IRAP team acknowledges support from the French space agency (Centre National des Etudes Spatiales; CNES; https://cnes.fr/fr) that funds activity of the space weather team in Toulouse (Solar-Terrestrial Observations and Modelling Service; STORMS; http://stormsservice.irap.omp.eu/). The work of L. Griton, A.P.R., and N.P. was funded by the ERC SLOW_SOURCE project (SLOW_SOURCE—DLV-819189). We acknowledge the NASA Parker Solar Probe Mission, the FIELDS team led S. D. Bale, and the SWEAP team led by J. Kasper for use of data. The FIELDS and SWEAP experiments on the Parker Solar Probe spacecraft were designed and developed under NASA contract NNN06AA01C.

Different states of the slow solar wind are identified from in-situ measurements by Parker Solar Probe (PSP) inside 50 solar radii from the Sun (Encounters 1, 2, 4, 5 and 6). At such distances the wind measured at PSP has not yet undergone significant transformation related to the expansion and propagation of the wind. We focus in this study on the properties of the quiet solar wind with no magnetic switchbacks. The Slow Solar Wind (SSW) states differ by their density, flux, plasma beta and magnetic pressure. PSP's magnetic connectivity established with Potential Field Source Surface (PFSS) reconstructions, tested against extreme ultraviolet (EUV) and white-light imaging, reveals the different states under study generally correspond to transitions from streamers to equatorial coronal holes. Solar wind simulations run along these differing flux tubes reproduce the slower and denser wind measured in the streamer and the more tenuous wind measured in the coronal hole. Plasma heating is more intense at the base of the streamer field lines rooted near the boundary of the equatorial hole than those rooted closer to the center of the hole. This results in a higher wind flux driven inside the streamer than deeper inside the equatorial hole.

International audience; The enrichment of coronal loops and the slow solar wind with elements that have low First Ionization Potential, known as the FIP effect, has often been interpreted as the tracer of a common origin. A current explanation for this FIP fractionation rests on the influence of ponderomotive forces and turbulent mixing acting at the top of the chromosphere. The implied wave transport and turbulence mechanisms are also key to wave-driven coronal heating and solar wind acceleration models. This work makes use of a shell turbulence model run on open and closed magnetic field lines of the solar corona to investigate with a unified approach the influence of magnetic topology, turbulence amplitude and dissipation on the FIP fractionation. We try in particular to assess whether there is a clear distinction between the FIP effect on closed and open field regions.

Brazzein is a small sweet-tasting protein found in the red berries of a West African evergreen shrub, Pentadiplandra brazzeana Baillon. Brazzein is highly soluble and stable over a large pH range and at high temperatures, which are characteristics that suggest its use as a natural sweetener. However, Pentadiplandra brazzeana culture is difficult at a large scale, limiting the natural source of brazzein. Heterologous expression of brazzein has been established in numerous systems, including bacteria, yeast, and transgenic plants. Brazzein requires four disulfide bonds to be active in eliciting an intense sweet taste, and the yeast Pichia pastoris appears to be one of the best options for obtaining functional brazzein in high quantities. Employing yeast secretion in the culture medium allows us to obtain fully active brazzein and facilitate purification later. To increase yeast secretion, we compared seven different signal peptides to successfully achieve brazzein secretion using the yeast P. pastoris. The brazzein proteins corresponding to these signal peptides elicited activation of the sweet taste receptor functionally expressed in a cellular assay. Among these tested signal peptides, three resulted in the secretion of brazzein at high levels.

Two states of the slow solar wind are identified from in-situ measurements by Parker Solar Probe (PSP) inside 50 solar radii from the Sun. At such distances the wind measured at PSP has not yet undergone significant transformation related to the expansion and propagation of the wind. We focus in this study on the properties of the quiet solar wind with no magnetic switchbacks. The two states differ by their plasma beta, flux and magnetic pressure. PSP's magnetic connectivity established with Potential Field Source Surface (PFSS) reconstructions, tested against extreme ultraviolet (EUV) and white-light imaging, reveals the two states correspond to a transition from a streamer to an equatorial coronal hole. The expansion factors of magnetic field lines in the streamer are 20 times greater than those rooted near the center of the coronal hole. The very different expansion rates of the magnetic field result in different magnetic pressures measured by PSP in the two plasma states. Solar wind simulations run along these differing flux tubes reproduce the slower and denser wind measured in the streamer and the more tenuous wind measured in the coronal hole. Plasma heating is more intense at the base of the streamer field lines rooted near the boundary of the equatorial hole than those rooted closer to the center of the hole. This results in a higher wind flux driven inside the streamer than deeper inside the equatorial hole., Comment: Accepted for publication in the Astrophysical Journal on Feb 4, 2021

Background: BI 765063 (OSE-172) is a humanised IgG4 monoclonal antibody which binds selectively to the V1 allele of Signal Regulatory Protein α [SIRPα] blocking the SIRPα/CD47 “don't eat me” pathway. Preclinical studies showed that SIRPα blockage led to macrophage and T-cell recruitment into tumor xenografts, and induced upregulation of chemokines, cytokines and adaptive immune function genes in human tumor explants (Gauttier et al., 2020). The goal of the biomarker analyses was to characterize the BI 765063 impact on peripheral blood immune cells (PBMCs) and the tumor microenvironment (TME). Methods: Fifty patients (26 V1/V1, 24 V1/V2) received BI 765063 IV from 0.02 mg/kg to 36 mg/kg every 3 weeks. Paired tumor biopsies were collected before and 2 weeks after first BI 765063 infusion. PBMCs were collected before, then 4 h, 1, 14, and 21 days after first infusion. BI 765063 receptor occupancy (RO) was determined on peripheral CD14+ monocytes. Immunophenotyping of PBMCs was performed by flow-cytometry. TME was analysed with a Brightplex® IHC panel including CD8+ T-cells, CD68+ macrophages, SIRPα, CD47, and PD-L1. Tumor gene expression profiling was performed using the Pan Cancer Immune gene set. Results: BI 765063 full RO saturation was achieved at trough levels (C2D1, pre-dose) in V1/V1 patients treated with doses of 6 mg/kg and higher, while V1/V2 patients showed a more heterogeneous RO ranging from 40-80%, reaching an apparent saturation at ≥ 12 mg/kg. An increase of activated CD80+/CD14+ and CD40+/CD14+ monocytes in PBMCs was observed at 24 h post-treatment in both, V1/V1 and V1/V2 patients. In paired tumor biopsies, IFNγ, MHCII antigen presentation gene pathways, and CCL7 transcripts appeared to be upregulated at C1D15 in patients with a systemic exposure of ≥ 100 µg/ml. One patient with hepatocellular carcinoma (HCC) and liver and lung metastases treated with BI 765063 monotherapy at 24 mg/kg achieved partial response (Champiat et al., ASCO, 2021). Baseline tumor biopsy of that patient showed that 66% of HCC tumor cells were CD47+ and 87% of CD68+ macrophages were SIRPα+. Furthermore, high levels of CD8+ T-cells were observed at baseline. At C1D15 increased CD68+ macrophage infiltration, sustained CD8 T-cell tumor accumulation and higher PD-L1 CPS (48% at baseline vs 75% at C1D15) were observed. Analysis of paired tumor biopsies in other patients showed that often, increased levels of tumor CD68+ macrophages were accompanied by CD8+ T-cell infiltration. Conclusion: This early biomarker analysis in patients with a wide range of solid tumors and treated with the first-in-class SIRPa inhibitor BI 765063 show encouraging signs of potentially mode-of-action related changes, both in peripheral blood and the TME. These early signals will be further evaluated in similar samples from the ongoing expansion cohorts in more homogeneous patient populations. Citation Format: Stephane Champiat, Philippe A. Cassier, Nuria Kotecki, Carlos Gomez-Roca, Aurélien Marabelle, Armelle Vinceneux, Christiane Jungels, Mabrouk Elgadi, Ralph Graeser, Thomas Vandewalle, Isabelle Girault, Nina Salabert-Le Guen, Nicolas Poirier, Bérangère Vasseur, Dominique Costantini, Claudia Fromond, Jean-Pierre Delord. Biomarker analyses from the Phase I clinical trial of the first-in-class SIRPa immune checkpoint inhibitor BI765063 in patients with advanced solid tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1993.

SummarySince the beginning of the COVID19 pandemics, an unprecedented research effort has been conducted to analyze the antibody responses in patients, and many trials based on passive immunotherapy — notably monoclonal antibodies — are ongoing. Twenty-one antibodies have entered clinical trials, 6 having reached phase 2/3, phase 3 or having received emergency authorization. These represent only the tip of the iceberg, since many more antibodies have been discovered and represent opportunities either for diagnosis purposes or as drug candidates. The main problem facing laboratories willing to develop such antibodies is the huge task of analyzing them and choosing the best candidate for exhaustive experimental validation. In this work we show how artificial intelligence-based methods can help in analyzing large sets of antibodies in order to determine in a few hours the best candidates in few hours. The MAbCluster method, which only requires knowledge of the amino acid sequences of the antibodies, allows to group the antibodies having the same epitope, considering only their amino acid sequences and their 3D structures (actual or predicted), and to infer some of their functional properties. We then use MAbTope to predict the epitopes for all antibodies for which they are not already known. This allows an exhaustive comparison of the available epitopes, but also gives a synthetic view of the possible combinations. Finally, we show how these results can be used to predict which antibodies might be affected by the different mutations arising in the circulating strains of the virus, such as the N501Y mutation that has started to spread in Great-Britain.

Brazzein is a small sweet-tasting protein found in the red berries of a West African evergreen shrub, Pentadiplandra brazzeana Baillon. Brazzein is highly soluble and stable over a large pH range and at high temperatures, which are characteristics that suggest its use as a natural sweetener. However, Pentadiplandra brazzeana culture is difficult at a large scale, limiting the natural source of brazzein. Heterologous expression of brazzein has been established in numerous systems, including bacteria, yeast, and transgenic plants. Brazzein requires four disulfide bonds to be active in eliciting an intense sweet taste, and the yeast Pichia pastoris appears to be one of the best options for obtaining functional brazzein in high quantities. Employing yeast secretion in the culture medium allows us to obtain fully active brazzein and facilitate purification later. To increase yeast secretion, we compared seven different signal peptides to successfully achieve brazzein secretion using the yeast P. pastoris. The brazzein proteins corresponding to these signal peptides elicited activation of the sweet taste receptor functionally expressed in a cellular assay. Among these tested signal peptides, three resulted in the secretion of brazzein at high levels.

Background Myeloid cells represent one of the most abundant immune cell types in solid tumors that impede myeloid phagocytosis by triggering ‘don’t eat me’ and ‘don’t find me’ signals. Recent literature demonstrates that C-type lectin receptors (CLRs) normally constrain immune cell–mediated tissue damage by suppressing myeloid cell activation and then promote tumor immune evasion. We previously identified the orphan (CLRs) CLEC-1 as over-expressed in situation of established immune tolerance and reported that CLEC-1 expression by dendritic cells (DCs) and macrophages is enhanced by TGFβ and tempers downstream T cells responses. Furthermore, we reported that CLEC-1 is highly expressed by myeloid cells purified from human tumor micro-environment significantly more expressed by suppressive macrophages. Methods As DCs and macrophages are professional phagocytes of dying/dead cell, we evaluated whether CLEC-1 could be a receptor of damaged cells in the phagocytosis. Results We found that CLEC-1 fusion protein, binds specifically to late apoptotic and secondary necrotic healthy or tumor cells induced by chemotherapy, radiation (UV, X-ray) or culture stress conditions. Importantly, we observed in vivo that CLEC-1 deficient mice, but not wild-type, eradicate MC38 colorectal tumors in combination with cytotoxic and immunogenic chemotherapy (eg. Cyclophosphamide. We then generated, screened and identified different anti-human Clec-1 antagonist monoclonal antibodies (mAbs) with the capacity to block the CLEC-1/CLEC-1L interaction. We discovered that various antagonist CLEC-1 mAbs, but not non-antagonist CLEC-1 control mAbs, increase the phagocytosis of CLEC-1L-positive human tumor cells by human CLEC-1 expressing TGFβ-polarized DCs or macrophages. Indeed, TGFβ-polarized DCs phagocytosed more efficiently Rituximab (anti-CD20 mAb)-opsonized Burkitt lymphoma cells (Raji) as well as bare NSCLC cells (A549) when CLEC-1 is antagonized by antibodies. Furthermore, macrophages more productively engulfed Rituximab-opsonized Raji cells as well in the context of CLEC-1 blockade (2–3 fold increase). Moreover, Cetuximab opsonized colon carcinoma cells (DLD-1; EGFR+) and Trastuzumab opsonized mammary carcinoma cells (SK-BR-3; Her2+) were likewise more phagocytosed by CLEC-1 blocked macrophages. Conclusions Altogether, these data indicate illustrate that CLEC-1 broadly inhibits tumor-cell phagocytosis and synergized with tumor-targeted cytotoxic monoclonal antibodies in both solid and hematological tumors.

All authors: Rouillard, A. P.; Pinto, R. F.; Vourlidas, A.; De Groof, A.; Thompson, W. T.; Bemporad, A.; Dolei, S.; Indurain, M.; Buchlin, E.; Sasso, C.; Spadaro, D.; Dalmasse, K.; Hirzberger, J.; Zouganelis, I.; Strugarek, A.; Brun, A. S.; Alexandre, M.; Berghmans, D.; Raouafi, N. E.; Wiegelmann, T.; Pagano, P.; Arge, C. N.; Nieves-Chinchilla, T.; Lavarra, M.; Poirier, N.; Amari, T.; Aran, A.; Andretta, V.; Antonucci, E.; Anastasiadis, A.; Auchère, F.; Bellot Rubio, L.; Nicula, B.; Bonnin, X.; Bouchemit, M.; Budnik, E.; Caminade, S.; Cecconi, B.; Carlyle, J.; Cernuda, I.; Davila, J. M.; Etesi, L.; Espinosa Lara, F.; Fedorov, A.; Fineschi, S.; Fludra, A.; Génot, V.; Georgoulis, M. K.; Gilbert, H. R.; Giunta, A.; Gomez-Herrero, R.; Guest, S.; Haberreiter, M.; Hassler, D.; Henney, C. J.; Howard, R. A.; Horbury, T. S.; Janvier, M.; Jones, S. I.; Kozarev, K.; Kraaikamp, E.; Kouloumvakos, A.; Krucker, S.; Lagg, A.; Linker, J.; Lavraud, B.; Louarn, P.; Maksimovic, M.; Maloney, S.; Mann, G.; Masson, A.; Müller, D.; Önel, H.; Osuna, P.; Orozco Suarez, D.; Owen, C. J.; Papaioannou, A.; Pérez-Suárez, D.; Rodriguez-Pacheco, J.; Parenti, S.; Pariat, E.; Peter, H.; Plunkett, S.; Pomoell, J.; Raines, J. M.; Riethmüller, T. L.; Rich, N.; Rodriguez, L.; Romoli, M.; Sanchez, L.; Solanki, S. K.; St Cyr, O. C.; Straus, T.; Susino, R.; Teriaca, L.; del Toro Iniesta, J. C.; Ventura, R.; Verbeeck, C.; Vilmer, N.; Warmuth, A.; Walsh, A. P.; Watson, C.; Williams, D.; Wu, Y.; Zhukov, A. N.-- Open Access article, published by EDP Sciences, under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited., Context. The Solar Orbiter spacecraft will be equipped with a wide range of remote-sensing (RS) and in situ (IS) instruments to record novel and unprecedented measurements of the solar atmosphere and the inner heliosphere. To take full advantage of these new datasets, tools and techniques must be developed to ease multi-instrument and multi-spacecraft studies. In particular the currently inaccessible low solar corona below two solar radii can only be observed remotely. Furthermore techniques must be used to retrieve coronal plasma properties in time and in three dimensional (3D) space. Solar Orbiter will run complex observation campaigns that provide interesting opportunities to maximise the likelihood of linking IS data to their source region near the Sun. Several RS instruments can be directed to specific targets situated on the solar disk just days before data acquisition. To compare IS and RS, data we must improve our understanding of how heliospheric probes magnetically connect to the solar disk. Aims. The aim of the present paper is to briefly review how the current modelling of the Sun and its atmosphere can support Solar Orbiter science. We describe the results of a community-led effort by European Space Agency's Modelling and Data Analysis Working Group (MADAWG) to develop different models, tools, and techniques deemed necessary to test different theories for the physical processes that may occur in the solar plasma. The focus here is on the large scales and little is described with regards to kinetic processes. To exploit future IS and RS data fully, many techniques have been adapted to model the evolving 3D solar magneto-plasma from the solar interior to the solar wind. A particular focus in the paper is placed on techniques that can estimate how Solar Orbiter will connect magnetically through the complex coronal magnetic fields to various photospheric and coronal features in support of spacecraft operations and future scientific studies. Methods. Recent missions such as STEREO, provided great opportunities for RS, IS, and multi-spacecraft studies. We summarise the achievements and highlight the challenges faced during these investigations, many of which motivated the Solar Orbiter mission. We present the new tools and techniques developed by the MADAWG to support the science operations and the analysis of the data from the many instruments on Solar Orbiter. Results. This article reviews current modelling and tool developments that ease the comparison of model results with RS and IS data made available by current and upcoming missions. It also describes the modelling strategy to support the science operations and subsequent exploitation of Solar Orbiter data in order to maximise the scientific output of the mission. Conclusions. The on-going community effort presented in this paper has provided new models and tools necessary to support mission operations as well as the science exploitation of the Solar Orbiter data. The tools and techniques will no doubt evolve significantly as we refine our procedure and methodology during the first year of operations of this highly promising mission. © 2020 A. P. Rouillard et al., Solar Orbiter is a joint ESA and NASA mission. A. Vourlidas' Solar Orbiter effort is supported by NRL grant N00173-16-1-G029. P. Pagano would like to thank D. H. Mackay and S. L. Yardley for their valuable contributions, the European Research Council (ERC) under the European Union Horizon 2020 research and innovation program (grant agreement No. 647214), and the DiRAC Data Centric system at Durham University, operated by the Institute for Computational Cosmology on behalf of the STFC DiRAC HPC Facility (http://www.dirac.ac.uk).This equipment was funded by a BIS National E-infrastructure capital grant ST/K00042X/1, STFC capital grant ST/K00087X/1, DiRAC Operations grant ST/K003267/1 and Durham University. DiRAC is part of the National E-Infrastructure. A. Rouillard, V. Genot, M. Janvier, Elie Soubrier, F. Auchere, E. Buchlin and E. Pariat acknowledge support from the French space agency (Centre National d'Etudes Spatiales; CNES; https://cnes.fr/fr) that funds activity in plasma physics data center (Centre de Donnees de la Physique des Plasmas; CDPP; http://cdpp.eu/) and the Multi Experiment Data and Operation Center (MEDOC; https://idoc.ias.u-psud.fr/MEDOC), and the space weather team in Toulouse (Solar-Terrestrial Observations and Modelling Service; STORMS; http://storms-service.irap.omp.eu/).This includes funding for Gaia-DEM, the data mining tools AMDA (http://amda.cdpp.eu/), CLWEB (clweb.cesr. fr/) and the propagation tool (http://propagationtool.cdpp.eu).R.Pinto, M. Lavarra, Y. Wu and A. Kouloumvakos acknowledge financial support from the ANR project SLOW_ SOURCE No. ANR-17-CE31-0006-01, ANR project COROSHOCK No. ANR-18-ERC1-0006-01 and FP7 HELCATS project https://www.helcats-fp7.eu/under the FP7 EU contract number 606692. A. Warmuth acknowledges the support by DLR under grant No. 50 QL 0001. The STEREO SECCHI data are produced by a consortium of RAL (UK), NRL (USA), LMSAL (USA), GSFC (USA), MPS (Germany), CSL (Belgium), IOTA (France) and IAS (France). The ACE data were obtained from the ACE science center. The WIND data were obtained from the Space Physics Data Facility. Javier Rodriguez-Pacheco acknowledges Spanish Project: FEDER/MCIU-AEI/Project ESP2017-88436-R.

Vertebrate odorant-binding proteins (OBPs) are small soluble proteins abundantly secreted in the olfactory mucus of many animal species, including humans. Vertebrate OBPs reversibly bind odorant molecules with micromolar range affinities. Although their physiological role is not clearly understood, OBPs are proposed to carry airborne odorants toward membrane olfactory receptors through the nasal mucus. Measurements of odorant-OBP interactions and structural studies require a large amount of pure OBPs devoid of ligands. The bacterial expression system is the first choice for expressing vertebrate OBPs used in our laboratory and others. This system generally produces OBPs in large amounts without major problems. In this chapter, we describe the milligram-scale production of recombinant pig OBP1 (pOBP1) in E. coli. The different steps of expression and purification are presented and discussed. Protocols for secondary structures investigation by circular dichroism and binding properties of the recombinant protein are also provided. More generally, these approaches can be used to produce and characterize any vertebrate OBPs for use in functional and structural studies.

International audience; Odorant-binding proteins (OBPs) are small soluble proteins that are thought to transport hydrophobic odorants across the aqueous sensillar lymph to olfactory receptors. A recent study revealed that OBP28a, one of the most abundant Drosophila OBPs, is not required for odorant transport, but acts in buffering rapid odour variation in the odorant environment. To further unravel and decipher its functional role, we expressed recombinant OBP28a and characterized its binding specificity. Using a fluorescent binding assay, we found that OBP28a binds a restricted number of floral-like chemicals, including ß-ionone, with an affinity in the micromolar range. We solved the X-ray crystal structure of OBP28a, which showed extensive conformation changes upon ligand binding. Mutant flies genetically deleted for the OBP28a gene showed altered responses to ß-ionone at a given concentration range, supporting its essential role in the detection of specific compounds present in the natural environment of the fly.

International audience; We present Heliospheric Current Sheet (HCS) and Plasma Sheet (HPS) observations during Parker Solar Probe's (PSP) first orbit around the Sun. We focus on the eight intervals that display a true sector boundary (TSB; based on suprathermal electron pitch angle distributions) with one or several associated current sheets. The analysis shows that (1) the main density enhancements in the vicinity of the TSB and HCS are typically associated with electron strahl dropouts, implying magnetic disconnection from the Sun, (2) the density enhancements are just about twice that in the surrounding regions, suggesting mixing of plasmas from each side of the HCS, (3) the velocity changes at the main boundaries are either correlated or anticorrelated with magnetic field changes, consistent with magnetic reconnection, (4) there often exists a layer of disconnected magnetic field just outside the high-density regions, in agreement with a reconnected topology, (5) while a few cases consist of short-lived density and velocity changes, compatible with short-duration reconnection exhausts, most events are much longer and show the presence of flux ropes interleaved with higher-β regions. These findings are consistent with the transient release of density blobs and flux ropes through sequential magnetic reconnection at the tip of the helmet streamer. The data also demonstrate that, at least during PSP's first orbit, the only structure that may be defined as the HPS is the density structure that results from magnetic reconnection, and its by-products, likely released near the tip of the helmet streamer.

Recent analysis of high-cadence white-light images taken by the Solar-Terrestrial RElations Observatory (STEREO) near solar maximum has revealed that outflowing density structures are released in an ubiquitous manner in the solar wind. The present study investigates whether these density fluctuations could originate from the transient heating of the low corona observed during Coronal Bright Points (CBPs). We assume that part of the intense heating measured during CBPs occurs at the coronal base of open magnetic fields that channel the forming solar wind. We employ the solar wind model MULTI-VP to quantify the plasma compression induced by transient heating and investigate how the induced perturbation propagates to the upper corona. We show that for heating rates with statistics comparable to those observed during CBPs the compressive wave initially increases the local plasma density by a factor of up to 50$\%$ at 5 R$_\odot$. The wave expands rapidly beyond 30 solar radii and the local enhancement in density decreases beyond. Based on the occurrence rates of CBPs measured in previous studies, we impose transient heating events at the base of thousands of open magnetic field lines to study the response of the entire 3-D corona. The simulated density cubes are then converted into synthetic white-light imagery. We show that the resulting brightness variations occupy all position angles in the images on timescales of hours. We conclude that a significant part of the ubiquitous brightness variability of the solar corona could originate in the strong transient heating of flux tubes induced by CBPs., Comment: 15 pages, 10 figures, accepted for publication in The Astrophysical Journal

OBJECTIVE Primary Sjogren's syndrome (SS) is characterized by a lymphocytic infiltration of salivary glands (SGs) and the presence of an interferon (IFN) signature. SG epithelial cells (SGECs) play an active role in primary SS pathophysiology. We undertook this study to examine the interactions between SGECs and T cells in primary SS and the role of the interleukin-7 (IL-7)/IFN axis. METHODS Primary cultured SGECs from control subjects and patients with primary SS were stimulated with poly(I-C), IFNα, or IFNγ. T cells were sorted from blood and stimulated with IL-7. CD25 expression was assessed by flow cytometry. SG explants were cultured for 4 days with anti-IL-7 receptor (IL-7R) antagonist antibody (OSE-127), and transcriptomic analysis was performed using the NanoString platform. RESULTS Serum IL-7 level was increased in patients with primary SS compared to controls and was associated with B cell biomarkers. IL7R expression was decreased in T cells from patients with primary SS compared to controls. SGECs stimulated with poly(I-C), IFNα, or IFNγ secreted IL-7. IL-7 stimulation increased the activation of T cells, as well as IFNγ secretion. Transcriptomic analysis of SG explants showed a correlation between IL7 and IFN expression. Finally, explants cultured with anti-IL-7R antibody showed decreased IFN-stimulated gene expression. CONCLUSION These results suggest the presence of an IL-7/IFNγ amplification loop involving SGECs and T cells in primary SS. IL-7 was secreted by SGECs stimulated with type I or type II IFN and, in turn, activated T cells that secrete type II IFN. An anti-IL-7R antibody decreased the IFN signature in T cells in primary SS and could be of therapeutic interest.

The Wide-field Imager for Solar PRobe (WISPR) obtained the first high-resolution images of coronal rays at heights below 15 Rsun when Parker Solar Probe (PSP) was located inside 0.25 AU during the first encounter. We exploit these remarkable images to reveal the structure of coronal rays at scales that are not easily discernible in images taken from near 1 AU. To analyze and interpret WISPR observations which evolve rapidly both radially and longitudinally, we construct a latitude versus time map using full WISPR dataset from the first encounter. From the exploitation of this map and also from sequential WISPR images we show the presence of multiple sub-structures inside streamers and pseudo-streamers. WISPR unveils the fine-scale structure of the densest part of streamer rays that we identify as the solar origin of the heliospheric plasma sheet typically measured in situ in the solar wind. We exploit 3-D magneto-hydrodynamic (MHD) models and we construct synthetic white-light images to study the origin of the coronal structures observed by WISPR. Overall, including the effect of the spacecraft relative motion towards the individual coronal structures we can interpret several observed features by WISPR. Moreover, we relate some coronal rays to folds in the heliospheric current sheet that are unresolved from 1 AU. Other rays appear to form as a result of the inherently inhomogeneous distribution of open magnetic flux tubes. This work was funded by the European Research Council through the project SLOW_SOURCE - DLV-819189.

During its first solar encounter, the Parker Solar Probe (PSP) acquired unprecedented up-close imaging of a small Coronal Mass Ejection (CME) propagating in the forming slow solar wind. The CME originated as a cavity imaged in extreme ultraviolet that moved very slowly ($, 19 pages, 10 figures, to appear in ApJS, 2 animations of Figure 2 available at https://nuage.irap.omp.eu/index.php/s/8ynNotNVWU6PrtN and https://nuage.irap.omp.eu/index.php/s/dflQ94DKuMbiaTv

Depuis trente ans, les milieux d’altitude europeens constituent un cadre novateur de recherche sur les dynamiques des systemes pastoraux des espaces de montagne dans la longue duree. Les differentes equipes de recherche impliquees dans ces travaux se trouvent confrontees au probleme de l’acquisition de l’information primaire lors de la phase de prospection. Acceder a une information archeologique qui soit la plus complete possible dans un temps relativement court est l’enjeu de la phase de prospection en milieu d’altitude. Le programme de recherche TAHMM (Teledetection Archeologique en Haute et Moyenne Montagne) a pour objectif de reviser les methodes de detection des structures archeologiques de ces milieux en deployant differentes methodes non invasives, du satellite a l’analyse chimique des sols, pour mettre en place une approche integree multi-source, multi-echelle et multi-temporelle. Etablir de nouvelles procedures de prospection demande egalement de pousser plus avant l’analyse de l’information et souvent de sortir des schemas trop stricts des formats proprietaires. Le present article exposera les premiers resultats du travail de refonte de la chaine de traitement et d’analyse des images thermiques acquises par drone sur des sites pastoraux d’altitude en s’appuyant sur des tests realises en 2018.

The recent democratisation of UAVs and the progress made in sensor miniaturisation now make it possible to embark miniaturised LiDaRs on board mini-UAVs. This possibility opens the way to a more systematic use of Lidar technology for the survey of small forested areas and for the mapping of isolated archaeological sites. The paper presents a feedback from the Garonne valley where the acquisition of Lidar data by drone on three archaeological forested sites allows to discuss the variations in the extent and use of the current forested area. The quality of UAV-Lidar data is also compared to data acquired by plane.

Vertebrate odorant-binding proteins (OBPs) are small soluble proteins abundantly secreted in the olfactory mucus of many animal species, including humans. Vertebrate OBPs reversibly bind odorant molecules with micromolar range affinities. Although their physiological role is not clearly understood, OBPs are proposed to carry airborne odorants toward membrane olfactory receptors through the nasal mucus. Measurements of odorant-OBP interactions and structural studies require a large amount of pure OBPs devoid of ligands. The bacterial expression system is the first choice for expressing vertebrate OBPs used in our laboratory and others. This system generally produces OBPs in large amounts without major problems. In this chapter, we describe the milligram-scale production of recombinant pig OBP1 (pOBP1) in E. coli. The different steps of expression and purification are presented and discussed. Protocols for secondary structures investigation by circular dichroism and binding properties of the recombinant protein are also provided. More generally, these approaches can be used to produce and characterize any vertebrate OBPs for use in functional and structural studies.

International audience; Gurmarin is a highly specific sweet-taste suppressing protein in rodents that is isolated from the Indian plant Gymnemasylvestre. Gurmarin consists of 35 amino acid residues containing three intramolecular disulfide bridges that form a cystine knot. Here, we report the crystal structure of gurmarin at a 1.45 Å resolution and compare it with previously reported NMR solution structures. The atomic structure at this resolution allowed us to identify a very flexible region consisting of hydrophobic residues. Some of these amino acid residues had been identified as a putative binding site for the rat sweet taste receptor in a previous study. By combining alanine-scanning mutagenesis of the gurmarin molecule and a functional cell-based receptor assay, we confirmed that some single point mutations in these positions drastically affect sweet taste receptor inhibition by gurmarin.

Despite the favorable prognosis of B cell precursor acute lymphoblastic leukemia (BCP-ALL), relapse remains a clinical challenge. Antibody-based immunotherapies like Blinatumomab have improved BCP-ALL outcome, yet these can quickly turn ineffective due to CD19 antigen escape. Therefore, novel targeted immunotherapy options are urgently needed. We previously showed that high IL7R expression associates with adverse outcome in BCP-ALL patients and that IL7R targeting using laboratory-grade blocking antibodies significantly reduced leukemic burden in E2A-PBX1 and BCR-ABL patient derived xenografts (PDX) (Alsadeq et al., Blood, 2017; Abdelrasoul et al., Nat. Commun., 2020). Here we used the humanized monoclonal antibody OSE-127, a full IL7R antagonist which is not internalized by target cells and prevents IL7R heterodimerization and subsequent downstream signaling (Belarif et al., Nat. Commun. 2018, Belarif et al. JCI, 2019). Importantly, OSE-127 has recently been positively evaluated in a phase 1 study conducted in 63 healthy volunteers receiving either a single dose or two doses two weeks apart (EUDRACT 2018-001832-22). OSE-127 demonstrated an excellent safety and tolerability profile showing no signs of lymphopenia, cytokine release syndrome or T-cell compartment alterations. Based on these preclinical and clinical results, we investigated the anti-leukemic efficacy of OSE-127 immunotherapy in PDX models of BCP-ALL. First, to identify patients that may benefit from IL7R-immunotherapy, we measured IL7R surface expression in diagnostic BCP-ALL patient samples of different cytogenetic subgroups via flow cytometry. We detected IL7R-positivity (defined as ≥10% IL7R + BCP-ALL cells) in 52% (49/94) of patient samples including BCR-ABL + (25%; 5/20), TEL-AML1 + (41%; 7/17) and B-other (40%, 8/20) samples. Particularly high numbers of IL7R + patients were found in the E2A-PBX1 + and MLL-rearranged (MLLr) BCP-ALL subgroups (79%; 19/24 and 85%; 11/13 IL7R + patient samples, respectively). Next, we tested the efficacy of OSE-127 in NSG mice injected with PDX cells from two different E2A-PBX1 + patients (63.0% and 89.6% IL7R-positive blasts, respectively) modelling an MRD-situation by starting treatment one day after injection of PDX cells. While 100% of control animals developed overt leukemia, 0% of the OSE-127-treated animals developed the disease, with 10/10 and 7/10 mice per PDX-group being MRD-negative by the time of sacrifice. Of note, OSE-127-therapy was also effective in corresponding PDX models when therapy was initiated upon detection of 1% BCP-ALL cells in the peripheral blood, modelling an overt leukemia situation (P To investigate the efficacy of OSE-127 in a variety of BCP-ALL samples, we conducted a preclinical phase II-like study using PDX from different cytogenetic patient subgroups (8E2A-PBX1 +, including 1 matched diagnostic/relapse sample, 1 BCR-ABL, 1 MLLr and 1 ETV6-NTRK3). Two mice per PDX were injected, randomly assigned to control or OSE-127 treatment groups and therapy was initiated when >1% blasts were detected in the peripheral blood. Response to OSE-127 was detected in 9/11 (82%) PDX samples in vivo, including BCR-ABL +, MLLr and E2A-PBX1 +-relapse samples. Accordingly, OSE-127 immunotherapy led to a significant increase in median overall survival (85 days versus 55 days; P=0.0122). Of note, OSE-127 therapy response associated with IL7R expression levels on BCP-ALL cells and no significant downregulation of the IL7R antigen was observed upon OSE-127 treatment. Taken together, our data demonstrate that IL7R-targeting using OSE-127, which has already demonstrated a good safety profile in healthy volunteers, is an efficient approach for BCP-ALL immunotherapy and may be particularly beneficial for patients with high IL7R-expression and/or relapsed/refractory disease after CD19-directed therapy. Disclosures Lenk: OSE Immunotherapeutics: Research Funding. Baccelli: OSE Immunotherapeutics: Current Employment, Other: Author of patents related to anti-IL7R antibodies. Corallo: OSE Immunotherapeutics: Current Employment. Schrappe: SHIRE: Other: research support; JazzPharma: Honoraria, Other: research support; Servier: Honoraria, Other: research support; Novartis: Honoraria, Other: research support; SigmaTau: Other: research support; Amgen: Other: research support; Servier: Honoraria; Novartis: Honoraria; JazzPharma: Honoraria. Cario: Novartis: Other: Lecture Fee. Brüggemann: Incyte: Other: Advisory Board; Amgen: Other: Advisory Board, Travel support, Research Funding, Speakers Bureau; Janssen: Speakers Bureau. Poirier: OSE Immunotherapeutics: Current Employment, Other: Author of patents related to anti-IL7R antibodies. Schewe: OSE Immunotherapeutics: Research Funding; Bayer: Other: Advisory Board; SOBI: Other: Advisory Board; Jazz Pharmaceuticals: Other: Advisory Board.

BackgroundOSE2101 (Tedopi®) is an anticancer vaccine with HLA-A2+ restricted modified epitopes targeting five tumor-associated antigens (TAAs) frequently expressed in lung cancer (CEA, HER2, MAGE2, MAGE3, P53). Step-1 results of the phase III, randomized, open-label ATALANTE-1 study comparing Tedopi® vs standard treatment (SoC) showed a favorable benefit/risk of Tedopi® over SoC (HR 0.71 for overall survival OS) in HLA-A2+ NSCLC patients in 2nd or 3rd line treatment after progression on immune checkpoint blockers (ICB).1 We analyze available tumor biopsies at initial diagnosis from some patients treated with Tedopi® to determine the expression of the 5 TAAs and to identify other tumor factors associated with long-term survival.MethodsTumor biopsies were available for 8 HLA-A2+ (blood test) stage IV NSCLC patients included in the trial. Primary (ResultsHLA-class I was expressed in all tumor samples. IHC analysis revealed that P53, CEA and HER2 were expressed in 6/7, 5/7 and 0/7 patients, respectively. P53, CEA, HER2, MAGE2, and MAGE3 were detected at RNA level in 5/5 tested patients (table 1). IMMUNOSCORE® IC CD8/PDL1 analysis showed High/High, High/Low and Low/Low scores for 1/7, 1/7 and 5/7 patients, respectively. The High/High IMMUNOSCORE® with a pronounced CD8+ T-cell tumor infiltration was observed in the patient with PR. High percentage of tumor cells expressing P53 (69%–97%) and overexpression of genes associated with activated macrophages (TREM2, MARCO, SLC11A1, CHIT1, SERPINB2) were observed in the PR and SD patients. High IFN-gamma and Expanded Immune Gene Signature scores were observed in long-term survivor patients with secondary resistance to ICB, even after progressive disease.Abstract 366 Table 1Summary of clinical and translational dataCEACarcinoembryonic antigen; HER2: Human Epidermal Growth Factor Receptor-2; ICB: Immune checkpoint blocker; IHC: Immunohistochemistry; ND: Not determined; OS: Overall Survival; Patient ID: Patient identification; PDL1: Programmed death-ligand 1; PFS: Progression-free survival; ssGSEA: Single-sample Gene Set Enrichment Analysis. Blue bars = Length of overall survival; Green bars = Gene Signature upregulation; Red bars = Gene Signature downregulationConclusionsThis study shows that all HLA-A2+ patients (blood test), expressed HLA class I in the tumors at initial diagnosis. Transcriptomic data in the patients that benefited from Tedopi® showed activated macrophage pathway, high IFN-gamma and Expanded Immune Gene Signatures scores. These data will be validated on larger number of patients treated with Tedopi® after the step 2 analysis.AcknowledgementsWe thank Julie Le Boulicaut, François Montestruc and Constant Josse (eXYSTAT, Malakoff, France) for the statistical analysis, and HalioDx for the IHC and NanoString analysis.Trial RegistrationEudraCT number2015-003183-36; NCT number: NCT02654587ReferenceGiaccone, et al. Activity of OSE-2101 in HLA-A2+ non-small cell lung cancer (NSCLC) patients after failure to immune checkpoint inhibitors (ICI): step 1 results of phase III ATALANTE-1 randomised trial. ESMO meeting 2020, abstract #1260MO.Ethics ApprovalThe study protocol and its related documents (including the patient information and informed consent form) received approval from the Institutional Review Board (IRB), and the Competent Authority prior to study initiation.ConsentEach patient gave his/her written informed consent prior to study enrolment.

BackgroundC-type lectin receptors (CLRs) are powerful pattern recognition receptors shaping immune cell-mediated tissue damage by positively or negatively regulating myeloid cell functions and hence tumor elimination or evasion. We previously reported that the orphan CLR CLEC-1 expressed by dendritic cells (DCs) tempers T cell’s responses in vivo by limiting antigen cross-presentation by cDC1. Furthermore, we observed that CLEC-1 is highly expressed by myeloid cells purified from human tumor microenvironment, in particular tumor-associated macrophages.MethodsMacrophages were generated from monocytes of healthy volunteers for phagocytosis assays. MC38 and Hepa 1.6 murine tumor cells were implanted in Clec1a KO or KI mice for immunotherapeutic treatment evaluation.ResultsUsing newly developed anti-human CLEC-1 monoclonal antibodies (mAbs), we found that antagonist anti-CLEC-1 mAbs with the capacity to block CLEC-1/CLEC-1Ligand interaction, as opposed to non-antagonist CLEC-1 mAbs, increase the phagocytosis of CLEC-1Ligand-positive human tumor cells by human macrophages, in particular when opsonized by tumor-associated antigen mAbs (Rituximab, Cetuximab, Trastuzumab) or with anti-CD47 mAb (Magrolimab). In-vivo, CLEC-1 knock-out (KO) mice (n=19) display significant prolonged survival in monotherapy as compared to wild-type littermates (n=12) in an orthotopic hepatocellular carcinoma (HCC) model and anti-tumor memory responses was demonstrated by tumor rechallenge in cured mice. CLEC1 KO mice also illustrate significant eradication of MC38 colorectal tumors in combination with chemotherapy promoting CLEC-1Ligand expression by tumor cells (n=16 with Gemcitabine or n=11 with Cyclophosphamide). HCC tumor microenvironment analysis after 2 weeks of tumor implantation shows significantly higher number of CD8+ and memory CD8+ T cells with reduced PD1 expression in CLEC1 KO animals (n=16 versus n=12 for KO vs WT mice respectively). Finally, we recently generated human CLEC-1 knock-in mice expressing the extracellular human CLEC1 domain fused to the intracellular mouse CLEC1 tail and confirmed preclinical efficacy in vivo with anti-human CLEC1 antagonist mAb in monotherapy in the orthotopic HCC model.ConclusionsThese data illustrate that CLEC-1 inhibition represents a novel therapeutic target for immuno-oncology modifying T cell immune responses and tumor cell phagocytosis by macrophages.

Myeloid cells represent one of the most abundant immune cell types in solid tumors that impede myeloid phagocytosis by triggering “don't eat me” and “don't find me” signals. Recent literature demonstrates that C-type lectin receptors (CLRs) are powerful pattern recognition receptors shaping immune cell-mediated tissue damage by positively or negatively regulating myeloid cell functions and hence tumor elimination or evasion. We previously reported that the orphan CLR CLEC-1 expressed by dendritic cells (DCs) and macrophages (MPs) is enhanced by TGFβ and tempers downstream T cells responses. Furthermore, we observed that CLEC-1 is highly expressed by myeloid cells purified from the human tumor microenvironment, in particular, tumor-associated MPs. We evaluated whether CLEC-1 could also be a receptor for DAMPs and influences phagocytosis. We found that CLEC-1 fusion protein binds specifically to secondary necrotic healthy or tumor cells induced by chemotherapy, radiation (UV, X-ray), or culture stress conditions suggesting that ligands of CLEC-1 are generated upon stress and programmed cell death. Importantly, further to promising results in KO CLEC-1 mice, we observed in vivo that CLEC-1 deficient mice, in contrast to wild-type littermates, eradicate MC38 colorectal tumors in combination with cytotoxic and immunogenic chemotherapy. Importantly, disruption of CLEC-1 signaling by Fc-CLEC-1 fusion protein also promotes tumor eradication. We then generated and identified different anti-human CLEC-1 antagonist monoclonal antibodies (mAbs) with the capacity to block CLEC-1/CLEC-1L interaction. We developed innovative antagonist CLEC-1 mAbs which, in contrast to non-antagonist CLEC-1 control mAb, increase the phagocytosis of CLEC-1L-positive human tumor cells by human TGFβ-polarized DCs or MPs. Indeed, TGFβ-polarized DCs phagocytose more efficiently a NSCLC cell line (A549) as well as Rituximab (anti-CD20 mAb)-opsonized Burkitt lymphoma cells (Raji) when CLEC-1 is antagonized by Abs. Similarly, macrophages significantly more efficiently engulfed human tumors in the presence of CLEC-1 antagonist Abs, in particular when tumor cells were opsonized such as Rituximab-opsonized Raji cells, Cetuximab opsonized colon carcinoma cells (DLD-1; EGFR+) or Trastuzumab opsonized mammary carcinoma cells (SK-BR-3; Her2+). Importantly, we observed both in vitro and in vivo that DCs from Clec1a deficient mice cross-present more efficiently dead cell-associated antigens to CD8+ T cells (OT-1). We generated hCLEC-1 knock-in mice and in vivo preclinical evaluation of CLEC-1 blocking mAbs is ongoing. Altogether, these data illustrate that CLEC-1 broadly notably inhibits tumor-cell phagocytosis and synergized with tumor-targeted cytotoxic monoclonal antibodies in both solid and hematological tumors, and hampers DC antigen cross-presentation. Citation Format: Vanessa Gauttier, Sabrina Pengam, Marion Drouin, Javier Saenz, Bérangère Evrard, Kevin Biteau, Caroline Mary, Géraldine Teppaz, Ariane Desselle, Virginie Thépénier, Emmanuelle Wilhelm, Elise Chiffoleau, Nicolas Poirier. CLEC-1 is a novel myeloid immune checkpoint for cancer immunotherapy limiting tumor cells phagocytosis and tumor antigen cross-presentation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1636.

Chronic inflammation is associated with abnormal non-phlogistic clearance (efferocytosis) of apoptotic cells by macrophages and a defect of the resolution of inflammation pathways. The resolution of inflammation is an active immunological process mediated by specialized pro-resolving mediator (SPM) which target specific resolutive G-protein coupled receptors expressed by different immune cells and participate to the tissue homeostasis return after an injury. Defects in the clearance of (chemotherapy-induced) apoptotic tumor cell debris strengthens inflammation and has been associated with exacerbated tumor growth in several preclinical models. Proresolutive therapeutic approaches, such as using exogenous resolvin E1 (RvE1), the natural lipidic proresolutive ligand of the GPCR ChemR23, have been shown to dampen tumor-associated inflammation and to reduce tumor growth. Using single-cell RNA sequencing analysis, we found that ChemR23 is mainly expressed by tumor-associated macrophages in melanoma and lung cancers. Moreover, ChemR23 expression was barely expressed in non-inflamed tissues indicating a preferential expression on the inflamed or tumor site. We screened and identified an anti-ChemR23 mAb which induces RvE1-like Akt and ERK signaling in mouse and human macrophages and favors macrophage polarization towards a pro-resolutive phenotype. In vitro, the agonist antibody significantly increased the phagocytosis of apoptotic tumor primary mesothelioma cells by human tumor-polarized macrophages. In vivo, the agonist ChemR23 mAb accelerates the resolution of inflammation in an acute inflammatory model, as illustrated by a significant decrease of inflammatory macrophages and neutrophils infiltrates as compared to isotype control groups both in mice and non-human primate models. Similarly, ChemR23 triggering with the anti-ChemR23 mAb improved weight recovery, reduced diarrhea and decreased the development of colon neoplastic foci in a chronic colitis model coupled with the injection of a carcinogen agent. Furthermore, the administration of the anti-ChemR23 mAb as a monotherapy displayed some partial and complete antitumor responses in immunocompetent mouse models of subcutaneous colon carcinoma (MC38 and CT26) as well as orthotopic mesothelioma (AK7). Finally, using an orthotopic triple-negative breast cancer model (4T1), while we observed a limited impact on primary tumor growth, spontaneous metastasis development in the lung was significantly inhibited by the ChemR23 agonist mAb monotherapy. Our study reveals for the first time the therapeutic potential of triggering the proresolutive pathways using an anti-ChemR23 agonistic mAb to limit chronic inflammation in the tumor microenvironment and inhibit metastasis development. Citation Format: Vanessa Gauttier, Margot Lavy, Charlène Trilleaud, Kévin Biteau, Isabelle Girault, Lyssia Belarif, Géraldine Teppaz, Caroline Mary, Virginie Thépénier, Christophe Blanquart, Sophie Barillé-Nion, Nicolas Poirier. Triggering the resolution of inflammation with agonistic anti-ChemR23 antibody dampens inflammation-driven carcinogenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1766.

Although anti PD(L)-1 therapy lead to impressive responses, the majority of patient are unresponsive or fail to develop a durable response after treatment. Immunocytokines have the potential to strengthen PD-(L)1 therapy by promoting T cell survival, however, their clinical developments are limited by a shortened half-life and systemic toxicity. To redirect immunotherapy to tumor specific T cells, we propose to selectively deliver IL-7 to PD1+ T cells using a bispecific anti-PD1 fused to IL-7 (BiCKI®IL-7). Intratumoral progenitor exhausted CD8+ T cells (Tpex) coexpressing PD1/IL-7R+ were described as the key subset responsive to anti PD-(L)1; but their efficacies are transient as Tpex rapidly undergo apoptosis limiting the full efficacy of anti-PD-(L)1. BICKI®IL7 was specifically designed to preferentially reinvigorate effector functions of PD1+IL-7R+ tumor-specific T cells, to sustain long-term anti-tumor response and counteract immune resistance in refractory patients. BICKI®IL7 was created by the fusion of a flexible linker and human IL-7 to the Fc portion of a high affinity antagonist anti-PD1 antibody. BICKI®IL7 constructed with wild-type IL-7 exhibited a high binding and potency (pSTAT5 IL-7 signaling) irrespective of PD1 expression. As our goal is to selectively activate the IL-7 signaling into PD1+ T cells but not PD1 negative cells, we fused different IL-7 muteins having 2 to 30,000-fold lower affinity for CD127/CD132 complex. One BICKI®IL7 mutein construction was selected for its loss of potency on PD1- cells and its optimal cis-binding/redirection and cis-activity on PD1+ cells. Furthermore, BICKI®IL7 mutein fully antagonized the PD-L1/2 interactions and inhibitory signaling to a similar extent to the naked anti PD1. Interestingly, BICKI®IL7 mutein synergistically activates TCR signaling while IL-7+anti-PD1 combo has no additive effect. Using a chronic antigen stimulation model, we demonstrated that BICKI®IL-7 efficiently activates and sustains the proliferation of early and fully exhausted T cells, and also confirmed the synergy. In parallel, we also explored the effect of the BICKI®IL7 mutein on Tregs in coculture assay. BICKI®IL7 mutein preferentially stimulated CD8 Teff over Treg as opposed to IL-2 & IL-15 and, also abrogated Treg suppression by restoring Teff IFN-γ secretion and proliferation. In vivo, BICKI®IL-7 mutein enhanced anti-tumor responses in mesothelioma model and promoted the proliferation and generation of memory CD8 T cells to higher extent to anti-PD1 treatment. Taken together, our data validate the therapeutic potential of providing IL-7 signal to strengthen PD1 therapy and prevent immunoresistance by sustaining T cell response and overcoming Treg suppression. The bispecific BiCKI®IL-7 mutein can preferentially deliver and activate IL-7 into PD1+ tumor-specific T cells limiting the risk of I-O/I-O immunotoxicity. Citation Format: Aurore Morello, Justine Durand, Margaux Seite, Virginie Thepenier, Géraldine Teppaz, Emmanuelle Wilhelm, Arianne Desselle, Caroline Mary, Nicolas Poirier. Optimized antagonist anti-PD-1/IL-7 bispecific antibody to sustain exhausted T cell function and to disarm Treg suppressive activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 692.

International audience; Developing protocols aimed at inhibiting effector T cells would be key for the treatment of T cell-dependent autoimmune diseases including type 1 autoimmune diabetes (T1D) and multiple sclerosis (MS). While heme oxygenase-1 (HO-1) inducers are clinically approved drugs for non-immune-related diseases, they do have immunosuppressive properties when administered systemically in rodents. Here we show that HO-1 inducers inhibit antigen-specific effector T cells when injected intradermally together with the T cell cognate antigens in mice. This phenomenon was observed in both a CD8 þ T cell-mediated model of T1D and in a CD4 þ T cell-dependent MS model. Intradermal injection of HO-1 in-ducers induced the recruitment of HO-1 þ monocyte-derived dendritic cell (MoDCs) exclusively to the lymph nodes (LN) draining the site of intradermal injection. After encountering HO-1 þ MoDCs, effector T-cells exhibited a lower velocity and a reduced ability to migrate towards chemokine gradients resulting in impaired accumulation to the inflamed organ. Intradermal co-injection of a clinically approved HO-1 inducer and a specific antigen to non-human primates also induced HO-1 þ MoDCs to accumulate in dermal draining LN and to suppress delayed-type hypersensitivity. Therefore, in both mice and non-human primates, HO-1 inducers delivered locally inhibited effector T-cells in an antigen-specific manner, paving the way for repositioning these drugs for the treatment of immune-mediated diseases.

International audience; Field-walking survey, the historically oldest method of modern archaeology, has undergone little technical development in its field phase. However, the question of the validity and representativeness of the data obtained gave rise to many discussions. The arrival of GPS was a significant improvement in recording capabilities, but remains limited to in-situ recording and analysis. Systematic georeferencing of off-site material has been attempted, but remains experimental, due to the technical complexity it entails, but above all because of its low cost-efficiency in terms of time spent on the documented area. We present an application for mobile phones using the increasingly powerful GPS chips embedded by them. The application allows a real time recording of the trace of a prospector’s path and the artifacts that were observed and collected. The GIS operation of the data collected enables the validity of the measured corpus to be evaluated and an intra-parcel spatial analysis of the surface material to be developed, capable of objectifying the identification of sites, specifying their extent and morphology, while maintaining a good performance of field operations.; La prospection pédestre, historiquement la plus ancienne des méthodes de l’archéologie moderne, a connu peu d’évolutions techniques sur le terrain. Pourtant la question de la validité et de la représentativité des données obtenues a donné lieu à de nombreuses discussions. L’arrivée du GPS a constitué une amélioration sensible des possibilités d’enregistrement, mais reste limitée à celui-ci et à l’analyse intra-site. Le géoréférencement systématique des données recueillies hors-sites a été tenté mais il demeure expérimental, en raison de la complexité technique qu’il induit et surtout de sa faible rentabilité si l’on rapporte le temps passé à la surface documentée. Nous présentons une application pour téléphones mobiles exploitant les puces GPS de plus en plus performantes qu’ils embarquent. L’application permet d’enregistrer en temps réel la trace du parcours d’un prospecteur et les artefacts qu’il a pu observer et collecter. L’exploitation sous SIG des données recueillies permet d’évaluer la validité des corpus mesurés et de développer une analyse spatiale intra-parcellaire du mobilier de surface à même d’objectiver l’identification de sites, de préciser leur emprise et leur morphologie, tout en conservant un bon rendement des opérations de terrain.

Objective: The purpose of the study was to compare student performance and student satisfaction ratings for an introductory extremities radiology course taught using 2 different educational methods. Methods: One group of students was taught using a traditional face-to-face instruction method, and the other group received an integrative blended-learning approach. A multivariate analysis of scores on lecture and laboratory examinations was performed to detect differences in student performance between the 2 methods. An independent t test was performed to compare the final course averages between the 2 methods. χ2 Analysis was used to compare the distribution of letter grades and levels of satisfaction between the 2 groups. Results: Test scores were higher for the integrative approach than for the traditional face-to-face method (p < .05). However, the differences were not meaningful, as the greatest improvement in correct responses was only for 2 questions. Students appeared to be more satisfied with the integrative approach when compared to the traditional method (p < .05). Conclusion: Student satisfaction with the educational delivery methods in an introductory extremities radiology course using an integrative approach was greater than for the traditional face-to-face instruction method. Student performance was similar between the 2 cohorts.

Remote observations of the solar photospheric light scattered by electrons (the K-corona) and dust (the F-corona or zodiacal light) have been made from the ground during eclipses1 and from space at distances as small as 0.3 astronomical units2–5 to the Sun. Previous observations6–8 of dust scattering have not confirmed the existence of the theoretically predicted dust-free zone near the Sun9–11. The transient nature of the corona has been well characterized for large events, but questions still remain (for example, about the initiation of the corona12 and the production of solar energetic particles13) and for small events even its structure is uncertain14. Here we report imaging of the solar corona15 during the first two perihelion passes (0.16–0.25 astronomical units) of the Parker Solar Probe spacecraft13, each lasting ten days. The view from these distances is qualitatively similar to the historical views from ground and space, but there are some notable differences. At short elongations, we observe a decrease in the intensity of the F-coronal intensity, which is suggestive of the long-sought dust free zone9–11. We also resolve the fine-scale plasma structure of very small eruptions, which are frequently ejected from the Sun. These take two forms: the frequently observed magnetic flux ropes12,16 and the predicted, but not yet observed, magnetic islands17,18 arising from the tearing-mode instability in the current sheet. Our observations of the coronal streamer evolution confirm the large-scale topology of the solar corona, but also reveal that, as recently predicted19, streamers are composed of yet smaller substreamers channelling continual density fluctuations at all visible scales. Observations of the solar corona by the Parker Solar Probe reveal evidence for the predicted dust-free zone and confirm that streamers comprise smaller substreamers that channel continuous multiscale density fluctuations.

Animals need to detect in the food essential amino acids that they cannot synthesize. We found that the odorant binding protein OBP19b, which is highly expressed in Drosophila melanogaster taste sensilla, is necessary for the detection of several amino acids including the essential l-phenylalanine. The recombinant OBP19b protein was produced and characterized for its binding properties: it stereoselectively binds to several amino acids. Using a feeding-choice assay, we found that OBP19b is necessary for detecting l-phenylalanine and l-glutamine, but not l-alanine or D-phenylalanine. We mapped the cells expressing OBP19b and compared the electrophysiological responses of a single taste sensillum to several amino acids: OBP19b mutant flies showed a reduced response compared to control flies when tested to preferred amino acids, but not to the other ones. OBP19b is well conserved in phylogenetically distant species suggesting that this protein is necessary for detection of specific amino acids in insects., Karen Rihani et al. demonstrate that fruit flies need an odorant-binding protein OBP19b, which is highly expressed in taste sensilla, to prefer select amino acids such as essential l-phenylalanine. This study provides insights into the mechanisms by which insects ensure their dietary intake of essential amino acids.

Dans le cadre du programme ANR Archeostraits, une cartographie de l'habitat du site archeologique de la Silla del Papa (sud de l'Espagne) a ete entreprise. Elle est rendue complexe par la topographie combinant de forts deniveles et une vegetation persistante. De plus, les bâtiments de la fin de l'âge du fer sont ancres a plusieurs niveaux dans les parois de barres rocheuses, ce qui rend inoperante une analyse architecturale traditionnelle a partir de planimetries en 2D. L'analyse 3D a plusieurs echelles, de l'environnement au sondage, demande une adaptation des outils d'acquisition 3D a cet espace. Trois technologies presentes au sein du laboratoire Traces generant des nuages de points denses ont ete utilisees.

Nephrotoxicity of calcineurin inhibitors and uncontrolled effector function of alloreactive T lymphocytes are main drivers of transplant dysfunctions. T lymphocytes either directly damage tissues or indirectly promote inflammation and antibody responses. Beside inhibitors of calcium-dependent pathways and antimetabolites, modulators of T-cell costimulation are elected pharmacological tools to enable interference with immune-mediated transplant dysfunctions. CD28 and CTLA-4 are major costimulatory and coinhibitory cell surface signaling molecules interacting with CD80/86, known to be critically important for immune response of committed T cells and regulation. Initial bench to beside translation, 2 decades ago, resulted in the development of belatacept CTLA-4 fused with an immunoglobulin Fc domain, a biologic inhibiting interaction of both CD28 and CTLA-4 with CD80/86. Despite proven effectiveness in inhibiting alloimmune responses, clinical use of belatacept in kidney transplantation revealed a substantially high incidence of acute, cell-mediated rejection. The cause of belatacept-resistant graft rejection was allocated to elevated pretransplant frequencies of CD28 memory T cells. Owing to different requirements in CD28 costimulatory and CTLA-4 coinhibitory signals to control naive and memory T cells, selective antagonists of CD28-CD80/86 interactions have been developed on the rationale that preservation of CTLA-4-mediated regulatory mechanisms would result in a better control of alloreactivity and would represent a regulatory T-cell-compatible immunosuppression. After the successful testing of selective CD28 antagonists in First In Human studies, this review delineates how this shift in paradigm performed in preclinical transplantation models and evaluates its clinical potential.

2623 Background: BI 765063 is a humanized IgG4 monoclonal antibody antagonist of SIRPα (Signal Regulatory Protein α), which blocks the “don't eat me” signal of the SIRPα/CD47 axis, a critical innate immune checkpoint. SIRPα is expressed on myeloid cells. BI 765063 binds to the V1 SIRPα allele with high affinity and to the V2 SIRPα allele with low affinity. BI 765063 lacks SIRPγ binding to preserve T-cell activation. We report results of the completed BI 765063 monotherapy dose escalation in patients with advanced solid tumors. Methods: This study involves a step 1 dose escalation to determine the dose-limiting toxicities (DLT) and maximum tolerated dose (MTD), then a step 2 dose-confirmation expansion at recommended phase 2 dose. In Step 1, BI 765063 ascending doses, given IV every 3 weeks, were tested using a Bayesian Logistic Regression Model (BLRM) approach with overdose control. The endpoints were safety, pharmacokinetics, receptor occupancy (RO) in peripheral CD14+ monocytes and efficacy (RECIST 1.1). Results: Fifty patients (26 V1/V1, 24 V1/V2) received at least one dose of BI 765063. The most frequent tumors were ovarian (9), colorectal (8), lung (5), breast (4), melanoma (3), and kidney (3). No DLTs were reported up to the highest dose tested. MTD was not reached. The most frequent related adverse events were infusion related reaction (IRR) (46%), fatigue (12%), headache (10%), arthralgia and diarrhea (8% each). All related adverse events were mild to moderate, except one case of IRR Grade 3. No related anemia nor thrombocytopenia were observed. BI 765063 showed dose proportional exposure and full RO saturation in Cycle 1 after the fourth dose level. Clinical benefit was observed in 21/47 (45%) patients evaluable per RECIST 1.1. One patient with hepatocellular carcinoma (HCC) with liver and lung metastases and 7 prior lines of therapy showed a durable partial response maintained for 27 weeks treatment (ongoing). The baseline tumor biopsy of this patient showed high CD8 T-cell and macrophage infiltration. There was an increase in CD8 T-cell infiltration and activation on treatment. An increase in PD-L1 expression on tumor cells 2 weeks after first dosing was also observed. Analysis of paired tumor biopsies in other patients is ongoing. Conclusions: The first-in-class SIRPα inhibitor BI 765063 was well-tolerated, showed monotherapy activity, and sustained RO saturation. A durable partial response was observed in an advanced HCC patient. The on-treatment biopsy of the responder showed an increase in CD8 T-cell infiltration and activation. PD-L1 expression on tumor cells also increased. BI 765063 dose escalation in combination with ezabenlimab (anti-PD1 antibody) is ongoing. Clinical trial information: NCT03990233.