Bacterial outer membrane vesicles and vaccine applications

Vaccines based on outer membrane vesicles (OMV) were developed more than 20 years ago against Neisseria meningitidis serogroup B. These nano-sized structures exhibit remarkable potential for immunomodulation of immune responses and delivery of “self” meningococcal antigens or unrelated antigens incorporated into the vesicle structure. This paper reviews different applications in OMV Research and Development (R&D) and provides examples of OMV developed and evaluated at the Finlay Institute in Cuba. A Good Manufacturing Practice (GMP) process was developed at the Finlay Institute to produce OMV from N. meningitidis serogroup B (dOMVB) using detergent extraction. Subsequently, OMV from N. meningitidis, serogroup A (dOMVA), serogroup W (dOMVW) and serogroup X (dOMVX) were obtained using this process. More recently, the extraction process has also been applied effectively for obtaining OMV on a research scale from Vibrio cholerae (dOMVC), Bordetella pertussis (dOMVBP), Mycobacterium smegmatis (dOMVSM) and BCG (dOMVBCG). The immunogenicity of the OMV have been evaluated for specific antibody induction, and together with functional bactericidal and challenge assays in mice have shown their protective potential. dOMVB has been evaluated with non-self neisserial antigens, including with a herpes virus type 2 glycoprotein, ovalbumin and allergens. In conclusion, OMV are proving to be more versatile than first conceived and remain an important technology for development of vaccine candidates. - See more at: http://journal.frontiersin.org/Journal/10.3389/fimmu.2014.00121/abstract#sthash.MwqUyZQ1.dpu

An investigation of error correcting techniques for OMV data

Papers on the following topics are presented: considerations of testing the Orbital Maneuvering Vehicle (OMV) system with CLASS; OMV CLASS test results (first go around); equivalent system gain available from R-S encoding versus a desire to lower the power amplifier from 25 watts to 20 watts for OMV; command word acceptance/rejection rates for OMV; a memo concerning energy-to-noise ratio for the Viterbi-BSC Channel and the impact of Manchester coding loss; and an investigation of error correcting techniques for OMV and Advanced X-ray Astrophysics Facility (AXAF)

Nanopods: A New Bacterial Structure and Mechanism for Deployment of Outer Membrane Vesicles

Background: Bacterial outer membrane vesicles (OMV) are packets of periplasmic material that, via the proteins and other molecules they contain, project metabolic function into the environment. While OMV production is widespread in proteobacteria, they have been extensively studied only in pathogens, which inhabit fully hydrated environments. However, many (arguably most) bacterial habitats, such as soil, are only partially hydrated. In the latter, water is characteristically distributed as films on soil particles that are, on average thinner, than are typical OMV (ca. ≤10 nm water film vs. 20 to >200 nm OMV;). Methodology/Principal Findings: We have identified a new bacterial surface structure, termed a "nanopod", that is a conduit for projecting OMV significant distances (e.g., ≥6 µm) from the cell. Electron cryotomography was used to determine nanopod three-dimensional structure, which revealed chains of vesicles within an undulating, tubular element. By using immunoelectron microscopy, proteomics, heterologous expression and mutagenesis, the tubes were determined to be an assembly of a surface layer protein (NpdA), and the interior structures identified as OMV. Specific metabolic function(s) for nanopods produced by Delftia sp. Cs1-4 are not yet known. However, a connection with phenanthrene degradation is a possibility since nanopod formation was induced by growth on phenanthrene. Orthologs of NpdA were identified in three other genera of the Comamonadaceae family, and all were experimentally verified to form nanopods. Conclusions/Significance: Nanopods are new bacterial organelles, and establish a new paradigm in the mechanisms by which bacteria effect long-distance interactions with their environment. Specifically, they create a pathway through which cells can effectively deploy OMV, and the biological activity these transmit, in a diffusion-independent manner. Nanopods would thus allow environmental bacteria to expand their metabolic sphere of influence in a manner previously unknown for these organisms

Rendezvous radar for the orbital maneuvering vehicle

The Rendezvous Radar Set (RRS) was designed at Motorola's Strategic Electronics Division in Chandler, Arizona, to be a key subsystem aboard NASA's Orbital Maneuvering Vehicle (OMV). The unmanned OMV, which was under development at TRW's Federal Systems Division in Redondo Beach, California, was designed to supplement the Shuttle's satellite delivery, retrieval, and maneuvering activities. The RRS was to be used to locate and then provide the OMV with vectoring information to the target satellite (or Shuttle or Space Station) to aid the OMV in making a minimum fuel consumption approach and rendezvous. The OMV development program was halted by NASA in 1990 just as parts were being ordered for the RRS engineering model. The paper presented describes the RRS design and then discusses new technologies, either under development or planned for development at Motorola, that can be applied to radar or alternative sensor solutions for the Automated Rendezvous and Capture problem

OMV mission simulator

The Orbital Maneuvering Vehicle (OMV) will be remotely piloted during rendezvous, docking, or proximity operations with target spacecraft from a ground control console (GCC). The real-time mission simulator and graphics being used to design a console pilot-machine interface are discussed. A real-time orbital dynamics simulator drives the visual displays. The dynamics simulator includes a J2 oblate earth gravity model and a generalized 1962 rotating atmospheric and drag model. The simulator also provides a variable-length communication delay to represent use of the Tracking and Data Relay Satellite System (TDRSS) and NASA Communications (NASCOM). Input parameter files determine the graphics display. This feature allows rapid prototyping since displays can be easily modified from pilot recommendations. A series of pilot reviews are being held to determine an effective pilot-machine interface. Pilots fly missions with nominal to 3-sigma dispersions in translational or rotational axes. Console dimensions, switch type and layout, hand controllers, and graphic interfaces are evaluated by the pilots and the GCC simulator is modified for subsequent runs. Initial results indicate a pilot preference for analog versus digital displays and for two 3-degree-of-freedom hand controllers

Collision avoidance for CTV: Requirements and capabilities

Cargo transfer vehicle (CTV) operations near Space Station Freedom will require positive collision avoidance maneuver (CAM) capability to preclude any change of collision, even in the event of CTV failures. The requirements for CAM are discussed, and the CAM design approach and design of the Orbiting Maneuvering Vehicle (OMV) are reviewed; this design met requirements for OMV operation near the Space Station, provided a redundant collision avoidance maneuver capability. Significant portions of the OMV CAM design should be applicable to CTV. The key features of the OMV design are summarized and related to the CTV mission design to that of OMV's. CAM is a defined sequence of events executed by the CTV to place the vehicle in a safe position relative to a target such as the Space Station. CAM can be performed through software commands to the propulsion system, or through commands pre-stored in hardware. Various techniques for triggering CAM are considered, and the risks associated with CAM enable and execution in phases are considered. OMV CAM design features both hardware and software CAM capability, with analyses conducted to assess the ability to meet the collision-free requirement during all phases of the mission

The Orbital Maneuvering Vehicle Training Facility visual system concept

The purpose of the Orbital Maneuvering Vehicle (OMV) Training Facility (OTF) is to provide effective training for OMV pilots. A critical part of the training environment is the Visual System, which will simulate the video scenes produced by the OMV Closed-Circuit Television (CCTV) system. The simulation will include camera models, dynamic target models, moving appendages, and scene degradation due to the compression/decompression of video signal. Video system malfunctions will also be provided to ensure that the pilot is ready to meet all challenges the real-world might provide. One possible visual system configuration for the training facility that will meet existing requirements is described

Design and implementation of a ground control console prototype for OMV

One of the primary uses of the Flat Floor facility of The Marshall Flight Center is to perform autonomous and teleoperated docking of vehicles similar to NASA's Orbital Maneuvering Vehicle (OMV). Using both the air-bearing vehicle and the 8 DOF Dynamic Overhead Target Simulator (DOTS) in tandem, even the most difficult OMV docking problems can be simulated. The OMV was planned to be flown from the ground by a pilot via teleoperation. Specifications were developed for a prototype Ground Control Console (GCC) from where the vehicle will be flown. In order for testing of the OMV to occur at Flat Floor facility, this GCC must be replicated. The project was divided into three primary tasks: (1) the design and development of the video display; (2) the design and development of the switch panel using the micro switch programmable display pushbutton switches; and (3) the design and development of the hand controllers. A final task is to perform system integration on the results of these three primary tasks

Preparation and Evaluation of a New Lipopolysaccharide-based Conjugate as a Vaccine Candidate for Brucellosis

Objectives: Development of an efficacious vaccine against brucellosis has been a challenge for scientists for many years. At present, there is no licensed vaccine against human brucellosis. To overcome this problem, currently, antigenic determinants of Brucella cell wall such as Lipopolysaccharide (LPS) are considered as potential candidates to develop subunit vaccines. Methods: In this study, Brucella abortus LPS was used for conjugation to Neisseria meningitidis serogroup B outer membrane vesicle (OMV) as carrier protein using carbodiimide and adipic acid-mediated coupling and linking, respectively. Groups of eight BALB/c mice were injected subcutaneously with 10μg LPS alone, combined LPS+OMV and conjugated LPS-OMV on 0 days, 14 days, 28 days and 42 days. Anti-LPS IgG was measured in serum. Results: The yield of LPS to OMV in LPS-OMV conjugate was 46.55, on the basis of carbohydrate content. The ratio for LPS to OMV was 4.07. The LPS-OMV conjugate was the most immunogenic compound that stimulated following the first injection with increased IgG titer of ~5-fold and ~1.3-fold higher than that produced against LPS and LPS in noncovalent complex to OMV (LPS+OMV), respectively. The highest anti-LPS IgG titer was detected 2 weeks after the third injection (Day 42) of LPS-OMV conjugate. The conjugated compound elicited higher titers of IgG than LPS+OMV, that showed a 100-120-fold rise of anti-LPS IgG in mice. Conclusion: These results indicate that our conjugated LPS-OMV can be used as a brucellosis vaccine, but further investigation is required. © 2014