A detailed study of charge collection efficiency has been performed on the Silicon Drift Detectors (SDD) of the ALICE experiment. Three different methods to study the collected charge as a function ...of the drift time have been implemented. The first approach consists in measuring the charge at different injection distances moving an infrared laser by means of micrometric step motors. The second method is based on the measurement of the charge injected by the laser at fixed drift distance and varying the drift field, thus changing the drift time. In the last method, the measurement of the charge deposited by atmospheric muons is used to study the charge collection efficiency as a function of the drift time. The three methods gave consistent results and indicated that no charge loss during the drift is observed for the sensor types used in 99% of the SDD modules mounted on the ALICE Inner Tracking System. The atmospheric muons have also been used to test the effect of the zero-suppression applied to reduce the data size by erasing the counts in cells not passing the thresholds for noise removal. As expected, the zero suppression introduces a dependence of the reconstructed charge as a function of drift time because it cuts the signal in the tails of the electron clouds enlarged by diffusion effects. These measurements allowed also to validate the correction for this effect extracted from detailed Monte Carlo simulations of the detector response and applied in the offline data reconstruction.
NK cells have therapeutic potential for a wide variety of human malignancies. However, because NK cells expand poorly in vitro, have limited life spans in vivo, and represent a small fraction of ...peripheral white blood cells, obtaining sufficient cell numbers is the major obstacle for NK-cell immunotherapy. Genetically-engineered artificial antigen-presenting cells (aAPCs) expressing membrane-bound IL-15 (mbIL15) have been used to propagate clinical-grade NK cells for human trials of adoptive immunotherapy, but ex vivo proliferation has been limited by telomere shortening. We developed K562-based aAPCs with membrane-bound IL-21 (mbIL21) and assessed their ability to support human NK-cell proliferation. In contrast to mbIL15, mbIL21-expressing aAPCs promoted log-phase NK cell expansion without evidence of senescence for up to 6 weeks of culture. By day 21, parallel expansion of NK cells from 22 donors demonstrated a mean 47,967-fold expansion (median 31,747) when co-cultured with aAPCs expressing mbIL21 compared to 825-fold expansion (median 325) with mbIL15. Despite the significant increase in proliferation, mbIL21-expanded NK cells also showed a significant increase in telomere length compared to freshly obtained NK cells, suggesting a possible mechanism for their sustained proliferation. NK cells expanded with mbIL21 were similar in phenotype and cytotoxicity to those expanded with mbIL15, with retained donor KIR repertoires and high expression of NCRs, CD16, and NKG2D, but had superior cytokine secretion. The mbIL21-expanded NK cells showed increased transcription of the activating receptor CD160, but otherwise had remarkably similar mRNA expression profiles of the 96 genes assessed. mbIL21-expanded NK cells had significant cytotoxicity against all tumor cell lines tested, retained responsiveness to inhibitory KIR ligands, and demonstrated enhanced killing via antibody-dependent cell cytotoxicity. Thus, aAPCs expressing mbIL21 promote improved proliferation of human NK cells with longer telomeres and less senescence, supporting their clinical use in propagating NK cells for adoptive immunotherapy.
Strong explosive eruptions of volcanoes throw out mixtures of gases and ash from high-pressure underground reservoirs. Investigating these subsurface reservoirs may help to forecast and characterize ...an eruption. In this study, we compare seismic tomography results with remote sensing and petrology data to identify deep and subaerial manifestations of pre-eruptive processes at Bezymianny volcano in Kamchatka shortly before its violent explosion on December 20, 2017. Based on camera networks we identify precursory rockfalls, and based on satellite radar data we find pre-eruptive summit inflation. Our seismic network recorded the P and S wave data from over 500 local earthquakes used to invert for a 3D seismic velocity distribution beneath Bezymianny illuminating its eruptive state days before the eruption. The derived tomography model, in conjunction with the presence of the high-temperature-stable SiO
polymorph Tridymite in juvenile rock samples , allowed us to infer the coexistence of magma and gas reservoirs revealed as anomalies of low (1.5) and high (2.0) Vp/Vs ratios, respectively, located at depths of 2-3 km and only 2 km apart. The reservoirs both control the current eruptive activity: while the magma reservoir is responsible for episodic dome growth and lava flow emplacements, the spatially separated gas reservoir may control short but powerful explosive eruptions of Bezymianny.
Lava domes grow by extrusions and intrusions of viscous magma often initiating from a central volcanic vent, and they are frequently defining the source region of hazardous explosive eruptions and ...pyroclastic density currents. Thus, close monitoring of dome building processes is crucial, but often limited to low data resolution, hazardous access, and poor visibility. Here, we investigated the 2016–2017 eruptive sequence of the dome building Bezymianny volcano, Kamchatka, with spot-mode TerraSAR-X acquisitions, and complement the analysis with webcam imagery and seismic data. Our results reveal clear morphometric changes preceding eruptions that are associated with intrusions and extrusions. Pixel offset measurements show >7 months of precursory plug extrusion, being locally defined and exceeding 30 m of deformation, chiefly without detected seismicity. After a short explosion, three months of lava dome evolution were characterised by extrusions and intrusion. Our data suggest that the growth mechanisms were significantly governed by magma supply rate and shallow upper conduit solidification that deflected magmatic intrusions into the uppermost parts of the dome. The integrated approach contributes significantly to a better understanding of precursory activity and complex growth interactions at dome building volcanoes, and shows that intrusive and extrusive growth is acting in chorus at Bezymianny volcano.
Abstract
The increase in number and intensity of earthquakes during a pre-eruptive crisis is the main basis of seismic volcano monitoring. However, a strong understanding of how these seismic signals ...relate to magmatic processes in the magma plumbing systems prior to volcanic eruptions is crucial for these efforts. Here we compare the characteristics of a seismo-volcanic crisis prior to the 2010–2013 explosive-extrusive eruption of Kizimen volcano, Kamchatka with the timescales of processes in the magma plumbing system. These timescales are inferred from the numerical modelling of iron-magnesium intracrystalline interdiffusion in 88 zoned orthopyroxene crystals from dacites and silica-rich andesites collected after the eruption. We find that the eruptible magmas were assembled rapidly during a magma mixing process beginning around 1.5 years before the eruption, which is well correlated with the onset of the seismic crisis. We conclude that the observed seismic re-activation marked the onset of magma mixing and led to destabilization of the reservoir, followed by the eruption.
Off the shelf T cell therapies for hematologic malignancies McCreedy, Bruce J.; Senyukov, Vladimir V.; Nguyen, Kim T.
Best practice & research. Clinical haematology,
June 2018, 2018-06-00, 20180601, Letnik:
31, Številka:
2
Journal Article
Recenzirano
Adoptive transfer of autologous CAR-T cells can induce durable remissions in patients with relapsed/refractory hematologic malignancies. However, multiple challenges exist for manufacturing CAR-T ...cells from patients with advanced disease including inability to manufacture a product, disease progression or death while waiting for the CAR-T product to be available, and heterogeneity among autologous CAR-T products that contributes to unpredictable and variable clinical activity. Healthy donor T cells can provide a source for production of universal CAR-T cells when combined with gene editing to prevent expression of endogenous TCRs and avoid generation of GvHD in HLA mismatched recipients. Additional gene edits can be included to impart resistance to immunosuppression or improve trafficking to tumor sites. Recent advances in cell manufacturing and analytics technology can provide for consistent batch to batch manufacturing of gene edited allogeneic CAR-T cells in sufficient quantity to treat thousands of patients when needed as off the shelf products.
The efficacy of most therapeutic monoclonal antibodies (mAbs) targeting tumor antigens results primarily from their ability to elicit potent cytotoxicity through effector-mediated functions. We have ...engineered the fragment crystallizable (Fc) region of the immunoglobulin G (IgG) mAb, HuM195, targeting the leukemic antigen CD33, by introducing the triple mutation Ser293Asp/Ala330Leu/Ile332Glu (DLE), and developed Time-lapse Imaging Microscopy in Nanowell Grids to analyze antibody-dependent cell-mediated cytotoxicity kinetics of thousands of individual natural killer (NK) cells and mAb-coated target cells. We demonstrate that the DLE-HuM195 antibody increases both the quality and the quantity of NK cell-mediated antibody-dependent cytotoxicity by endowing more NK cells to participate in cytotoxicity via accrued CD16-mediated signaling and by increasing serial killing of target cells. NK cells encountering targets coated with DLE-HuM195 induce rapid target cell apoptosis by promoting simultaneous conjugates to multiple target cells and induce apoptosis in twice the number of target cells within the same period as the wild-type mAb. Enhanced target killing was also associated with increased frequency of NK cells undergoing apoptosis, but this effect was donor-dependent. Antibody-based therapies targeting tumor antigens will benefit from a better understanding of cell-mediated tumor elimination, and our work opens further opportunities for the therapeutic targeting of CD33 in the treatment of acute myeloid leukemia.
•Fc-engineered mAb promotes NK cell ADCC via better activation, serial killing, and kinetic boosting at higher target cell densities.•Enhanced target killing also increased frequency of NK cell apoptosis, but this effect is donor-dependent.
Adoptive transfer of T cells expressing a CD19-specific chimeric antigen receptor (CAR) is being evaluated in multiple clinical trials. Our current approach to adoptive immunotherapy is based on a ...second generation CAR (designated CD19RCD28) that signals through a CD28 and CD3-ζ endodomain. T cells are electroporated with DNA plasmids from the Sleeping Beauty (SB) transposon/transposase system to express this CAR. Stable integrants of genetically modified T cells can then be retrieved when co-cultured with designer artificial antigen presenting cells (aAPC) in the presence of interleukin (IL)-2 and 21. Here, we reveal how the platform technologies of SB-mediated transposition and CAR-dependent propagation on aAPC were adapted for human application. Indeed, we have initiated clinical trials in patients with high-risk B-lineage malignancies undergoing autologous and allogeneic hematopoietic stem-cell transplantation (HSCT). We describe the process to manufacture clinical grade CD19-specific T cells derived from healthy donors. Three validation runs were completed in compliance with current good manufacturing practice for Phase I/II trials demonstrating that by 28 days of co-culture on γ-irradiated aAPC ∼10(10) T cells were produced of which >95% expressed CAR. These genetically modified and propagated T cells met all quality control testing and release criteria in support of infusion.
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