Dual targeting of CD19 and CD22 against B-ALL using a novel high-sensitivity aCD22 CAR

CAR T cells recognizing CD19 effectively treat relapsed and refractory B-ALL and DLBCL. However, CD19 loss is a frequent cause of relapse. Simultaneously targeting a second antigen, CD22, may decrease antigen escape, but is challenging: its density is approximately 10-fold less than CD19, and its large structure may hamper immune synapse formation. The characteristics of the optimal CD22 CAR are underexplored. We generated 12 distinct CD22 antibodies and tested CARs derived from them to identify a CAR based on the novel 9A8 antibody, which was sensitive to low CD22 density and lacked tonic signaling. We found no correlation between affinity or membrane proximity of recognition epitope within Ig domains 3–6 of CD22 with CART function. The optimal strategy for CD19/CD22 CART co-targeting is undetermined. Co-administration of CD19 and CD22 CARs is costly; single CARs targeting CD19 and CD22 are challenging to construct. The co-expression of two CARs has previously been achieved using bicistronic vectors. Here, we generated a dual CART product by co-transduction with 9A8-41BBζ and CAT-41BBζ (obe-cel), the previously described CD19 CAR. CAT/9A8 CART eliminated single- and double-positive target cells in vitro and eliminated CD19- tumors in vivo. CAT/9A8 CART is being tested in a phase I clinical study (NCT02443831)

miR-155 overexpressing monocytes resemble HLA highISG15 + synovial tissue macrophages from patients with rheumatoid arthritis and induce polyfunctional CD4+ T cell activation

MicroRNAs (miRs) are known to regulate pro-inflammatory effector functions of myeloid cells, and miR dysregulation is implicated in rheumatoid arthritis (RA), a condition characterized by inflammation and destruction of the joints. We showed previously that miR-155 is increased in myeloid cells in RA and induces pro-inflammatory activation of monocytes and macrophages; however, its role at the interface between innate and adaptive immunity was not defined. Here, RNA-sequencing revealed that overexpression of miR-155 in healthy donor monocytes conferred a specific gene profile which bears similarities to that of RA synovial fluid-derived CD14+ cells and HLAhighISG15+ synovial tissue macrophages, both of which are characterized by antigen-presenting pathways. In line with this, monocytes in which miR-155 was overexpressed, displayed increased expression of HLA-DR and both co-stimulatory and co-inhibitory molecules, and induced activation of polyfunctional T cells. Together, these data underpin the notion that miR-155-driven myeloid cell activation in the synovium contributes not only to inflammation but may also influence the adaptive immune response

Cell-specific bioorthogonal tagging of glycoproteins

Altered glycoprotein expression is an undisputed corollary of cancer development. Understanding these alterations is paramount but hampered by limitations underlying cellular model systems. For instance, the intricate interactions between tumour and host cannot be adequately recapitulated in monoculture of tumour-derived cell lines. More complex co-culture models usually rely on sorting procedures for proteome analyses and rarely capture the details of protein glycosylation. Here, we report a strategy termed Bio-Orthogonal Cell line-specific Tagging of Glycoproteins (BOCTAG). Cells are equipped by transfection with an artificial biosynthetic pathway that transforms bioorthogonally tagged sugars into the corresponding nucleotide-sugars. Only transfected cells incorporate bioorthogonal tags into glycoproteins in the presence of non-transfected cells. We employ BOCTAG as an imaging technique and to annotate cell-specific glycosylation sites in mass spectrometry-glycoproteomics. We demonstrate application in co-culture and mouse models, allowing for profiling of the glycoproteome as an important modulator of cellular function

Lazaroid (U-74389G) ameliorates lung injury due to lipid peroxidation and nitric oxide synthase-dependent reactive oxygen species generation caused by remote systematic ischemia-reperfusion following thoracoabdominal aortic occlusion

Introduction: Lung ischemia-reperfusion injury after thoracoabdominal aortic occlusion represents a major complication, which increases morbidity and mortality. In the present study we hypothesized that lazaroid U-74389G intravenous administration protects from lung ischemia-reperfusion injury through lipid peroxidation inhibition. Materials and methods: A total of 24 pigs were randomized in three groups. Group I (n = 8) underwent sham operation, group II (n = 8) underwent thoracoabdominal aortic occlusion for 45min and received placebo and group III (n = 8) received 3 doses of lazaroid (3 mg/kg) 60 and 30min before thoracoabdominal aortic occlusion and at 30min during thoracoabdominal aortic occlusion (duration 45min). Aortic occlusion was performed with aortic balloon-catheters under fluoroscopic guidance. All animals were sacrificed at the 7 t h postoperative day and lung specimens were harvested for molecular analysis. Results: mRNA levels of leukotrienes LB4 (LTB4R2), LC4 (LTC4S) and nitric oxide synthase (NOS) isoforms including iNOS, nNOS and eNOS were determined with real-time RT-qPCR. Nitric oxide can either induce (iNOS) or inhibit (nNOS and eNOS) lipid peroxidation based on its specific isoform origin. Group III showed significantly reduced mRNA levels of LTB4R2 (−63.7%), LTC4S (−35.9%) and iNOS (−60.2%) when compared with group II (P < 0.05, for all). The mRNA levels of nNOS was significantly increased (+37.4%), while eNOS was slightly increased (+2.1%) in group III when compared with group II (P < 0.05 and P = 0.467 respectively). Conclusion: Lazaroid U-74389G may represent an effective pharmacologic intervention in reducing lung ischemia-reperfusion injury following thoracoabdominal aortic occlusion. © 2018 IJS Publishing Group Lt

Bilateral transient osteoporosis of the hip. A case report and review of the literature

Transient osteoporosis of the hip is an uncommon, selflimiting condition that typically affects middle-aged men or, less frequently, women in the third trimester of pregnancy. It may begin spontaneously or after a minor trauma, and the main symptoms are acute pain, limping, and a reduced range of hip motion. Transient osteoporosis is also known as transient bone marrow edema and, when it affects more than one joint, as migratory osteoporosis syndrome. It often resembles osteonecrosis, but the two entities should not be confused because of their different prognoses and management. Magnetic resonance imaging plays an important role in primary diagnosis and reveals a characteristic pattern of bone marrow edema in the form of a diffuse low signal on T1 images and a high signal on T2 images. Conservative treatment is the gold standard and should be adhered to even in resistant cases. We describe a case of sequential bilateral transient hip osteoporosis due to continuous micro-traumas and review the current literature in terms of the diagnostic work-up, differential diagnosis and treatment of this rare entity. © 2018 CIC Edizioni Internazionali s.r.l. All rights reserved

Moderate intensity exercise training combined with inulin-propionate ester supplementation increases whole body resting fat oxidation in overweight women

BACKGROUND: Our previous work has shown that oral supplementation with inulin propionate ester (IPE) reduces intra-abdominal fat and prevents weight gain and that oral propionate intake enhances resting fat oxidation. The effects of IPE combined with exercise training on energy substrate utilisation are unknown. The aim of this study was to investigate the impact of 4-weeks IPE supplementation, in combination with a moderate intensity exercise training programme, on whole body fat oxidation and on plasma GLP-1 and PYY. METHODS: Twenty overweight healthy women participated in randomised parallel study and underwent 4 weeks of supervised exercise training either with IPE (EX/IPE group) or Placebo (EX/Placebo group) supplementation. Before and after the intervention participants conducted an experimental trial, which involved collection of expired gas and blood samples in the fasted state and during 7 h of the postprandial state. RESULTS: Within groups, the EX/IPE group significantly enhanced the amount of fat (Pre, 24.1 ± 1.2 g; Post, 35.9 ± 4.0 g, P  0.05). Comparing between groups, changes in the amount of fat oxidised were significantly (P < 0.05) different and a trend for difference was observed for amount of CHO oxidised (P = 0.06) and RER (P = 0.06). The interventions had no impact on fasting or postprandial plasma concentrations of GLP-1 and PYY. CONCLUSION: Moderate intensity exercise training programmes when combined with daily oral IPE supplementation may help overweight women to achieve increase in fat oxidation. The study was registered at clinicaltrials.gov as NCT04016350

Hybrid clouds for data-intensive, 5G-enabled IoT applications: An overview, key issues and relevant architecture

Hybrid cloud multi-access edge computing (MEC) deployments have been proposed as efficient means to support Internet of Things (IoT) applications, relying on a plethora of nodes and data. In this paper, an overview on the area of hybrid clouds considering relevant research areas is given, providing technologies and mechanisms for the formation of such MEC deployments, as well as emphasizing several key issues that should be tackled by novel approaches, especially under the 5G paradigm. Furthermore, a decentralized hybrid cloud MEC architecture, resulting in a Platform-as-a-Service (PaaS) is proposed and its main building blocks and layers are thoroughly described. Aiming to offer a broad perspective on the business potential of such a platform, the stakeholder ecosystem is also analyzed. Finally, two use cases in the context of smart cities and mobile health are presented, aimed at showing how the proposed PaaS enables the development of respective IoT applications. © 2019 by the authors. Licensee MDPI, Basel, Switzerland

An Edge-to-Cloud Virtualized Multimedia Service Platform for 5G Networks

The focus of research into 5G networks to date has been largely on the required advances in network architectures, technologies, and infrastructures. Less effort has been put on the applications and services that will make use of and exploit the flexibility of 5G networks built upon the concept of software-defined networking (SDN) and network function virtualization (NFV). Media-based applications are amongst the most demanding services, requiring large bandwidths for high audio-visual quality, low-latency for interactivity, and sufficient infrastructure resources to deliver the computational power for running the media applications in the networked cloud. This paper presents a novel service virtualization platform (SVP), called 5G-MEDIA SVP, which leverages the principles of NFV and SDN to facilitate the development, deployment, and operation of media services on 5G networks. The platform offers an advanced cognitive management environment for the provisioning of network services (NSs) and media-related applications, which directly link their lifecycle management with user experience as well as optimization of infrastructure resource utilization. Another innovation of 5G-MEDIA SVP is the integration of serverless computing with media intensive applications in 5G networks, increasing cost effectiveness of operation and simplifying development and deployment time. The proposed SVP is being validated against three media use cases: 1) immersive virtual reality 3-D gaming application; 2) remote production of broadcast content incorporating user generated contents; and 3) dynamically adaptive content distribution networks for the intelligent distribution of ultrahigh definition content. The preliminary results of the 5G-MEDIA SVP platform evaluation are compared against current practice and show that the proposed platform provides enhanced functionality for the operators and infrastructure owners, while ensuring better NS performance to service providers and end users