Low Cost Tuberculosis Vaccine Antigens in Capsules: Expression in Chloroplasts, Bio-Encapsulation, Stability and Functional Evaluation In Vitro

Tuberculosis (TB) caused by Mycobacterium tuberculosis is one of the leading fatal infectious diseases. The development of TB vaccines has been recognized as a major public health priority by the World Health Organization. In this study, three candidate antigens, ESAT-6 (6kDa early secretory antigenic target) and Mtb72F (a fusion polyprotein from two TB antigens, Mtb32 and Mtb39) fused with cholera toxin B-subunit (CTB) and LipY (a cell wall protein) were expressed in tobacco and/or lettuce chloroplasts to facilitate bioencapsulation/oral delivery. Site-specific transgene integration into the chloroplast genome was confirmed by Southern blot analysis. In transplastomic leaves, CTB fusion proteins existed in soluble monomeric or multimeric forms of expected sizes and their expression levels varied depending upon the developmental stage and time of leaf harvest, with the highest-level of accumulation in mature leaves harvested at 6PM. The CTB-ESAT6 and CTB-Mtb72F expression levels reached up to 7.5% and 1.2% of total soluble protein respectively in mature tobacco leaves. Transplastomic CTB-ESAT6 lettuce plants accumulated up to 0.75% of total leaf protein. Western blot analysis of lyophilized lettuce leaves stored at room temperature for up to six months showed that the CTB-ESAT6 fusion protein was stable and preserved proper folding, disulfide bonds and assembly into pentamers for prolonged periods. Also, antigen concentration per gram of leaf tissue was increased 22 fold after lyophilization. Hemolysis assay with purified CTB-ESAT6 protein showed partial hemolysis of red blood cells and confirmed functionality of the ESAT-6 antigen. GM1-binding assay demonstrated that the CTB-ESAT6 fusion protein formed pentamers to bind with the GM1-ganglioside receptor. The expression of functional Mycobacterium tuberculosis antigens in transplastomic plants should facilitate development of a cost-effective and orally deliverable TB booster vaccine with potential for long-term storage at room temperature. To our knowledge, this is the first report of expression of TB vaccine antigens in chloroplasts

Status of the emittance transfer experiment emtex

In order to improve the injection efficiency of the round UNILAC heavy ion beam into the asymmetric acceptance of the SIS18 it would be of great advantage to decreasethe horizontal emittance by a so called emittance transferto the vertical plane. In this contribution the present statusof the emittance transfer experiment EMTEX at GSI will be reported. A short introduction about the theoretical background of the technique will be given, while the mainpart is dedicated to the practical solutions setting up a testbeam line at GSI. Finally, the results of a first commissioning beam time will be presented. The scheduled beam time to apply the emittance transfer technique foreseen in spring 2014 had to be shifted to calendar week 26 in 2014, just after this conference as some components have not been delivered in time by the contractor. The results and comparison to the theoretical predictions you may find in later publications

Characterization of potential biomarkers of reactogenicity of licensed antiviral vaccines: randomized controlled clinical trials conducted by the BIOVACSAFE consortium

Funding text The authors are grateful for the vital contributions of the participating study volunteers, clinicians, nurses, and laboratory technicians at the Surrey study site. The work by Roberto Leone, laboratory technician at Humanitas Clinical and Research Center, is gratefully acknowledged. Finally, they thank Ellen Oe (GSK) for scientific writing assistance. The research leading to these results has received support from the Innovative Medicines Initiative Joint Undertaking under grant agreement n°115308, resources of which are composed of financial contribution from the European Union’s Seventh Framework Programme (FP7/2007–2013) and EFPIA companies’ in-kind contribution. The contribution of the European Commission to the Advanced Immunization Technologies (ADITEC) project (grant agreement n° 280873) is also gratefully acknowledged. Publisher Copyright: © 2019, The Author(s).Biomarkers predictive of inflammatory events post-vaccination could accelerate vaccine development. Within the BIOVACSAFE framework, we conducted three identically designed, placebo-controlled inpatient/outpatient clinical studies (NCT01765413/NCT01771354/NCT01771367). Six antiviral vaccination strategies were evaluated to generate training data-sets of pre-/post-vaccination vital signs, blood changes and whole-blood gene transcripts, and to identify putative biomarkers of early inflammation/reactogenicity that could guide the design of subsequent focused confirmatory studies. Healthy adults (N = 123; 20–21/group) received one immunization at Day (D)0. Alum-adjuvanted hepatitis B vaccine elicited vital signs and inflammatory (CRP/innate cells) responses that were similar between primed/naive vaccinees, and low-level gene responses. MF59-adjuvanted trivalent influenza vaccine (ATIV) induced distinct physiological (temperature/heart rate/reactogenicity) response-patterns not seen with non-adjuvanted TIV or with the other vaccines. ATIV also elicited robust early (D1) activation of IFN-related genes (associated with serum IP-10 levels) and innate-cell-related genes, and changes in monocyte/neutrophil/lymphocyte counts, while TIV elicited similar but lower responses. Due to viral replication kinetics, innate gene activation by live yellow-fever or varicella-zoster virus (YFV/VZV) vaccines was more suspended, with early IFN-associated responses in naïve YFV-vaccine recipients but not in primed VZV-vaccine recipients. Inflammatory responses (physiological/serum markers, innate-signaling transcripts) are therefore a function of the vaccine type/composition and presence/absence of immune memory. The data reported here have guided the design of confirmatory Phase IV trials using ATIV to provide tools to identify inflammatory or reactogenicity biomarkers.Peer reviewe

Test of the REX-RFQ and status of the front part of the REX-ISOLDE linac

For REX-ISOLDE (Radioactive beam EXperiments at ISOLDE/CERN), a test beamline is built up at the Garching Accelerator Lab. to perform He$^{1+}$-experiments with the RFQ, the matching (rebunching) section between RFQ and IH-DT-linac, the IH-structure and several electrostatic lenses of the REX-ISOLDE-mass separator. In a first step, the beamline is conceived for tests with the RFQ. This paper presents the parameters and the status of the REX-RFQ, the experimental setup and the particle dynamics simulations with the COSY infinity code for beam injection and beam analysis. Furthermore it shows the design and status of the mass separator, the IH- structure and the buncher section. (5 refs)

HITRAP: A facility at GSI for highly charged ions

An overview and status report of the new trapping facility for highly charged ions at the Gesellschaft fuer Schwerionenforschung is presented. The construction of this facility started in 2005 and is expected to be completed in 2008. Once operational, highly charged ions will be loaded from the experimental storage ring ESR into the HITRAP facility, where they are decelerated and cooled. The kinetic energy of the initially fast ions is reduced by more than fourteen orders of magnitude and their thermal energy is cooled to cryogenic temperatures. The cold ions are then delivered to a broad range of atomic physics experiments.Comment: 8 pages, 11 figure