Studies on processing, particle formation, and immunogenicity of the HIV-1 gag gene product: a possible component of a HIV vaccine

Antigens in a particulate conformation were shown to be highly immunogenic in mammals. For this reason, the particle forming capacity of derivatives of the HIV-1 group specific core antigen p55 gag was assayed and compared dependent on various expression systems: recombinant bacteria, vaccinia- and baculoviruses were established encoding the entire core protein p55 either in its authentic sequence or lacking the myristylation consensus signal. Moreover, p55 gag was expressed in combination with the protease (p55-PR) or with the entire polymerase (p55-pol), respectively. Budding of 100-160 nm p55 core particles, resembling immature HIV-virions, was observed in the eucaryotic expression systems only. In comparison to the vaccinia virus driven expression of p55 in mammalian cells, considerably higher yields of particulate core antigen were obtained by infection of Spodoptera frugiperda (Sf9) insect cells with the recombinant Autographa californica nuclear polyhedrosis (AcMNPV) baculovirus. Mutation of the NH2-terminal myristylation signal sequence prevented budding of the immature core particles. Expression of the HIV p55-PR gene construct by recombinant baculovirus resulted in complete processing of the p55 gag precursor molecule in this system. The introduction of an artificial frameshift near the natural frameshift site resulted in constitutive expression of the viral protease and complete processing of p55, both in Escherichia coli and in vaccinia virus infected cells. Interestingly, significant processing of p55 resembling that of HIV infected H9 cells could also be achieved in the vaccinia system by fusing the entire pol gene to the gag gene. Moreover, processing was not found to be dependent on amino-terminal myristylation of the gag procursor molecule, which is in contrast to observations with type C and type D retrovirus. However, complete processing of p55 into p24, p17, p9 and p6 abolished particle formation. Purified immature HIV-virus like particles were highly immunogenic in rabbits, leading to a strong humoral immune response after immunization. Empty immature p55 gag particles represent a noninfectious and attractive candidate for a basic vaccine component

T-cell and NK-cell infiltration into solid tumors: a key limiting factor for efficacious cancer immunotherapy.

Cancer immunotherapy has great promise, but is limited by diverse mechanisms used by tumors to prevent sustained antitumor immune responses. Tumors disrupt antigen presentation, T/NK-cell activation, and T/NK-cell homing through soluble and cell-surface mediators, the vasculature, and immunosuppressive cells such as myeloid-derived suppressor cells and regulatory T cells. However, many molecular mechanisms preventing the efficacy of antitumor immunity have been identified and can be disrupted by combination immunotherapy. Here, we examine immunosuppressive mechanisms exploited by tumors and provide insights into the therapies under development to overcome them, focusing on lymphocyte traffic

Interplay between microstructural evolution and tribo-chemistry during dry sliding of metals

Understanding the microstructural and tribo-chemical processes during tribological loading is of utmost importance to further improve the tribological behavior of metals. In this study, the friction, wear and tribo-chemical behavior of Ni with different initial microstructures (nanocrystalline, bi-modal, coarse-grained) is investigated under dry sliding conditions. In particular, the interplay be-tween frictional response, microstructural evolution and tribo-oxidation is considered. Friction tests are carried out using ball-on-disk experiments with alumina balls as counter-bodies, varying the load between 1 and 5 N. The microstructural evolution as well as the chemical reactions beneath the samples’ surface is investigated by means of cross-sections. The samples with finer microstructures show a faster run-in and lower maximum values of the coefficient of friction (COF) which can be attributed to higher oxidation kinetics and a higher hardness. It is observed that with increasing sliding cycles, a stable oxide layer is formed. Furthermore, initially coarse-grained samples show grain refinement, whereas initially finer microstructures undergo grain coarsening converging towards the same superficial grain size after 2,000 sliding cycles. Consequently, the experimental evidence supports that, irrespective of the initial microstructure, after a certain deformation almost identical steady-state COF values for all samples are achieved

T Cells Bearing a Chimeric Antigen Receptor against Prostate-Specific Membrane Antigen Mediate Vascular Disruption and Result in Tumor Regression.

Aberrant blood vessels enable tumor growth, provide a barrier to immune infiltration, and serve as a source of protumorigenic signals. Targeting tumor blood vessels for destruction, or tumor vascular disruption therapy, can therefore provide significant therapeutic benefit. Here, we describe the ability of chimeric antigen receptor (CAR)-bearing T cells to recognize human prostate-specific membrane antigen (hPSMA) on endothelial targets in vitro as well as in vivo. CAR T cells were generated using the anti-PSMA scFv, J591, and the intracellular signaling domains: CD3ζ, CD28, and/or CD137/4-1BB. We found that all anti-hPSMA CAR T cells recognized and eliminated PSMA(+) endothelial targets in vitro, regardless of the signaling domain. T cells bearing the third-generation anti-hPSMA CAR, P28BBζ, were able to recognize and kill primary human endothelial cells isolated from gynecologic cancers. In addition, the P28BBζ CAR T cells mediated regression of hPSMA-expressing vascular neoplasms in mice. Finally, in murine models of ovarian cancers populated by murine vessels expressing hPSMA, the P28BBζ CAR T cells were able to ablate PSMA(+) vessels, cause secondary depletion of tumor cells, and reduce tumor burden. Taken together, these results provide a strong rationale for the use of CAR T cells as agents of tumor vascular disruption, specifically those targeting PSMA. Cancer Immunol Res; 3(1); 68-84. ©2014 AACR

Results of the MTLRS-1 upgrade

In this report, the results of the upgrade of the German Modular Transportable Laser Ranging System MTLRS-1 are summarized. A short description of the new components and their influence on the system accuracy is given. It is shown, that the single shot accuracy of the MTLRS-1 has been improved from 5 cm to 1 cm

Development of an acoustic transceiver for the KM3NeT positioning system

[EN] In this paper we describe an acoustic transceiver developed for the KM3NeT positioning system. The acoustic transceiver is composed of a commercial free flooded transducer, which works mainly in the 20-40 kHz frequency range and withstands high pressures (up to 500 bars). A sound emission board was developed that is adapted to the characteristics of the transducer and meets all requirements: low power consumption, high intensity of emission, low intrinsic noise, arbitrary signals for emission and the capacity of acquiring the receiving signals with very good timing precision. The results of the different tests made with the transceiver in the laboratory and shallow sea water are described, as well as, the activities for its integration in the Instrumentation Line of the ANTARES neutrino telescope and in a NEMO tower for the in situ tests. © 2012 Elsevier B.V. All rights reserved.This work has been supported by the Ministerio de Ciencia e Innovacion (Spanish Government), Project references FPA2009-13983-C02-02, ACI2009-1067, AIC10-D-00583, and Consolider-Ingenio Multidark (CSD2009-00064). It has also been funded by Generalitat Valenciana, Prometeo/2009/26, and the European 7th Framework Programme, Grant no. 212525.Larosa, G.; Ardid Ramírez, M.; Llorens Alvarez, CD.; Bou Cabo, M.; Martínez Mora, JA.; Adrián Martínez, S.; KM3NeT Consortium (2013). Development of an acoustic transceiver for the KM3NeT positioning system. Nuclear Instruments and Methods in Physics Research Section A: Accelerators, Spectrometers, Detectors and Associated Equipment. 725:215-218. https://doi.org/10.1016/j.nima.2012.11.167S21521872

Spin Correlations in e+e−e^{+}e^{-} Pair Creation by Two-Photons and Entanglement in QED

Spin correlations of $e^{+}e^{-}$ pair productions of two colliding photons are investigated and explicit expressions for their corresponding probabilities are derived and found to be \textit{energy} (speed) dependent, for initially \textit{linearly} and \textit{circularly polarized} photons, different from those obtained by simply combining the spins of the relevant particles, for initially \textit{polarized} photons. These expressions also depend on the angles of spin of $e^{+}$ (and/or of $e^{-}$), for initially {\it linearly polarized} photons, but not for {\it circularly polarized} photons, as a function of the energy. It is remarkable that these explicit results obtained from quantum field theory show a clear violation of Bell's inequality of Local Hidden Variables theories at all {\it energies} beyond that of the threshold one for particle production, in support of quantum field theory in the relativistic regime. We hope that our explicit expression will lead to experiments, of the type described in the bulk of this paper, which can monitor energy (and speed) in polarization correlation experiments.Comment: 12 pages, 4 figure