Generation of functional CD8+ T Cells by human dendritic cells expressing glypican-3 epitopes

<p>Abstract</p> <p>Background</p> <p>Glypican 3 (GPC-3) is an oncofoetal protein that is expressed in most hepatocellular carcinomas (HCC). Since it is a potential target for T cell immunotherapy, we investigated the generation of functional, GPC-3 specific T cells from peripheral blood mononuclear cells (PBMC).</p> <p>Methods</p> <p>Dendritic cells (DC) were derived from adherent PBMC cultured at 37°C for 7 days in X-Vivo, 1% autologous plasma, and 800 u/ml GM-CSF plus 500 u/ml IL-4. Immature DC were transfected with 20 μg of <it>in vitro </it>synthesised GPC-3 mRNA by electroporation using the Easy-ject plus system (Equibio, UK) (300 V, 150 μF and 4 ms pulse time), or pulsed with peptide, and subsequently matured with lipopolysaccharide (LPS). Six predicted GPC-3 peptide epitopes were synthesized using standard f-moc technology and tested for their binding affinity to HLA-A2.1 molecules using the cell line T2.</p> <p>Results</p> <p>DC transfected with GPC-3 mRNA but not control DC demonstrated strong intracellular staining for GPC-3 and <it>in vitro </it>generated interferon-gamma expressing T cells from autologous PBMC harvested from normal subjects. One peptide, GPC-3_522-530FLAELAYDL, fulfilled our criteria as a naturally processed, HLA-A2-restricted cytotoxic T lymphocyte (CTL) epitope: i) it showed high affinity binding to HLA-A2, in T2 cell binding assay; ii) it was generated by the MHC class I processing pathway in DC transfected with GPC-3 mRNA, and iii) HLA-A2 positive DC loaded with the peptide stimulated proliferation in autologous T cells and generated CTL that lysed HLA-A2 and GPC-3 positive target cells.</p> <p>Conclusions</p> <p>These findings demonstrate that electroporation of GPC-3 mRNA is an efficient method to load human monocyte-derived DC with antigen because <it>in vitro </it>they generated GPC-3-reactive T cells that were functional, as shown by interferon-gamma production. Furthermore, this study identified a novel naturally processed, HLA-A2-restricted CTL epitope, GPC-3_522-530FLAELAYDL, which can be used to monitor HLA-A2-restricted CTL responses in patients with HCC. Further studies are required to investigate whether anti-GPC-3 immunotherapy has a role in the treatment of GPC-3 dependent tumours, such as HCC.</p

Improvement of Secondary Sedimentation Tanks in the Presence of a Baffle

Source: ICHE Conference Archive - https://mdi-de.baw.de/icheArchiv

Navigation Doppler Lidar for Autonomous Ground, Aerial, and Space Vehicles

A Doppler lidar instrument has been developed and demonstrated for providing critical vector velocity and altitude/range data for autonomous precision navigation. Utilizing advanced component technologies, this lidar can be adapted to different types of vehicles

RNA Nanoparticles for Brain Tumor Treatment

The presently-disclosed subject matter relates to an artificial RNA nanostructure molecule and method to treat brain tumor in a subject. More particularly, the presently disclosed subject matter relates to a RNA nanostructure containing a multiple branched RNA nanoparticle, a brain tumor targeting module, and an effective amount of a therapeutic agent. Further, the presently disclosed subject matter relates to a method of using the RNA nanostructure composition to treat brain tumor in a subject having or at risk of having brain tumor

LIDAR-Aided Inertial Navigation with Extended Kalman Filtering for Pinpoint Landing

In support of NASA s Autonomous Landing and Hazard Avoidance Technology (ALHAT) project, an extended Kalman filter routine has been developed for estimating the position, velocity, and attitude of a spacecraft during the landing phase of a planetary mission. The proposed filter combines measurements of acceleration and angular velocity from an inertial measurement unit (IMU) with range and Doppler velocity observations from an onboard light detection and ranging (LIDAR) system. These high-precision LIDAR measurements of distance to the ground and approach velocity will enable both robotic and manned vehicles to land safely and precisely at scientifically interesting sites. The filter has been extensively tested using a lunar landing simulation and shown to improve navigation over flat surfaces or rough terrain. Experimental results from a helicopter flight test performed at NASA Dryden in August 2008 demonstrate that LIDAR can be employed to significantly improve navigation based exclusively on IMU integration

Flight Testing ALHAT Precision Landing Technologies Integrated Onboard the Morpheus Rocket Vehicle

A suite of prototype sensors, software, and avionics developed within the NASA Autonomous precision Landing and Hazard Avoidance Technology (ALHAT) project were terrestrially demonstrated onboard the NASA Morpheus rocket-propelled Vertical Testbed (VTB) in 2014. The sensors included a LIDAR-based Hazard Detection System (HDS), a Navigation Doppler LIDAR (NDL) velocimeter, and a long-range Laser Altimeter (LAlt) that enable autonomous and safe precision landing of robotic or human vehicles on solid solar system bodies under varying terrain lighting conditions. The flight test campaign with the Morpheus vehicle involved a detailed integration and functional verification process, followed by tether testing and six successful free flights, including one night flight. The ALHAT sensor measurements were integrated into a common navigation solution through a specialized ALHAT Navigation filter that was employed in closed-loop flight testing within the Morpheus Guidance, Navigation and Control (GN&C) subsystem. Flight testing on Morpheus utilized ALHAT for safe landing site identification and ranking, followed by precise surface-relative navigation to the selected landing site. The successful autonomous, closed-loop flight demonstrations of the prototype ALHAT system have laid the foundation for the infusion of safe, precision landing capabilities into future planetary exploration missions

Recombinant λ-phage nanobioparticles for tumor therapy in mice models

Lambda phages have considerable potential as gene delivery vehicles due to their genetic tractability, low cost, safety and physical characteristics in comparison to other nanocarriers and gene porters. Little is known concerning lambda phage-mediated gene transfer and expression in mammalian hosts. We therefore performed experiments to evaluate lambda-ZAP bacteriophage-mediated gene transfer and expression in vitro. For this purpose, we constructed recombinant λ-phage nanobioparticles containing a mammalian expression cassette encoding enhanced green fluorescent protein (EGFP) and E7 gene of human papillomavirus type 16 (λ-HPV-16 E7) using Lambda ZAP- CMV XR vector. Four cell lines (COS-7, CHO, TC-1 and HEK-239) were transduced with the nanobioparticles. We also characterized the therapeutic anti-tumor effects of the recombinant λ-HPV-16 E7 phage in C57BL/6 tumor mice model as a cancer vaccine. Obtained results showed that delivery and expression of these genes in fibroblastic cells (COS-7 and CHO) are more efficient than epithelial cells (TC-1 and HEK-239) using these nanobioparticles. Despite the same phage M.O.I entry, the internalizing titers of COS-7 and CHO cells were more than TC-1 and HEK-293 cells, respectively. Mice vaccinated with λ-HPV-16 E7 are able to generate potent therapeutic antitumor effects against challenge with E7- expressing tumor cell line, TC-1 compared to group treated with the wild phage. The results demonstrated that the recombinant λ-phages, due to their capabilities in transducing mammalian cells, can also be considered in design and construction of novel and safe phage-based nanomedicines

Open-Loop Flight Testing of COBALT GN&C Technologies for Precise Soft Landing

A terrestrial, open-loop (OL) flight test campaign of the NASA COBALT (CoOperative Blending of Autonomous Landing Technologies) platform was conducted onboard the Masten Xodiac suborbital rocket testbed, with support through the NASA Advanced Exploration Systems (AES), Game Changing Development (GCD), and Flight Opportunities (FO) Programs. The COBALT platform integrates NASA Guidance, Navigation and Control (GN&C) sensing technologies for autonomous, precise soft landing, including the Navigation Doppler Lidar (NDL) velocity and range sensor and the Lander Vision System (LVS) Terrain Relative Navigation (TRN) system. A specialized navigation filter running onboard COBALT fuzes the NDL and LVS data in real time to produce a precise navigation solution that is independent of the Global Positioning System (GPS) and suitable for future, autonomous planetary landing systems. The OL campaign tested COBALT as a passive payload, with COBALT data collection and filter execution, but with the Xodiac vehicle Guidance and Control (G&C) loops closed on a Masten GPS-based navigation solution. The OL test was performed as a risk reduction activity in preparation for an upcoming 2017 closed-loop (CL) flight campaign in which Xodiac G&C will act on the COBALT navigation solution and the GPS-based navigation will serve only as a backup monitor