Le principe de subsidiarité au sens du droit de la Convention Européenne des Droits de l'Homme

As part of the European Convention, the principle of subsidiarity is marked by ambiguity. Originating from judges, it has not been formally defined in legal texts nor in any related jurisprudence and is therefore characterized by a semantic and legal indeterminacy. Featuring both a procedural and a tangible dimension, it remains a fundamental principle of European law. On the one hand, it structures, the system’s organization and operations. On the other hand, it guides the court of Strasbourg’s interpretation and review. Naturally ambiguous but still guiding European law, the principle of subsidiarity is presented as a flexible and malleable principle, and therefore adaptable. The analysis of its implementation in the European jurisprudence highlights its inconsistency and variability at the free disposal of the court of Strasbourg. However, in the light of the current context of an unprecedented saturation of the system and therefore of an urgent reassertion of its subsidiary nature, a fluctuating use of the principle is assessed differently to make it not a guarantee of national liberties, but a guide for them. As a driving force for increased accountability of member states, the principle of subsidiarity appears to be essential to the judicial policy of the European court.Le principe de subsidiarité, en droit de la Convention européenne, est un principe empreint d’ambiguïtés. D’origine prétorienne, il ne fait l’objet d’aucune définition formelle dans les textes ou la jurisprudence afférente et se voit ainsi marqué d’une indétermination sémantique et juridique certaine. Doté d’une double dimension à la fois procédurale et matérielle, il n’en reste pas moins un principe fondamental du droit européen, un principe qui structure, d’une part, l’organisation et le fonctionnement même du système et guide, d’autre part, l’interprétation et le contrôle du juge de Strasbourg. Parce que naturellement ambigu mais néanmoins directeur du droit européen, le principe de subsidiarité se présente donc comme un principe souple et malléable, par conséquent, adaptable. L’analyse de sa mise en oeuvre dans la jurisprudence européenne met ainsi en exergue l’inconstance et la variabilité d’application du principe, à la libre disposition du juge strasbourgeois. Mais analysée à la lumière du contexte actuel d’un engorgement sans précédent du système et donc d’une réaffirmation urgente de son caractère subsidiaire, l’utilisation fluctuante du principe s’apprécie sous un jour nouveau, visant à faire de la subsidiarité un principe non plus garant des libertés nationales mais désormais source d’un encadrement de celles-ci. Parce que moteur d’une responsabilisation accrue des Etats membres, le principe de subsidiarité apparaît donc comme une pièce maîtresse de la politique jurisprudentielle du juge européen

Topology of Schwann cells and sympathetic innervation along preglomerular vessels: A confocal microscopic study in protein S100B/EGFP transgenic mice

International audienceSchwann cells (Sc), associated axons, and nearby vascular endothelium constitute a functional trilogy of major importance during the development and regrowth of peripheral vascular nerves. The goal of the present study is to provide a technique of triple fluorescence confocal imaging of these cell types along renal preglomerular vessels. We took advantage of a protein S100B/EGFP transgenic mouse to visualize Sc. The endothelium was labeled with an intravenous injection of fluorescently tagged lectin, and after tissue processing, adrenergic nerves were revealed with an antibody against the marker protein synaptophysin. As a validation step, we found that EGFP-positive perivascular cells with prominent cell bodies and extensive, multidirectional cell processes were protein S100B positive. They were identified as Sc and indirectly assumed to be unmyelinated Sc. By contrast, we found strong EGFP expression in proximal epithelial cells and in the epithelium lining thin limbs of Henle. This epithelial fluorescence was not associated with immunoreactive protein S100B and thus corresponded to ectopic EGFP expressions in this mouse strain. Sc were organized in bundles or as a meshwork surrounding the preglomerular vasculature from arcuate arteries to afferent arterioles. No Sc were detected in the medulla. Although most Sc were closely apposed to adrenergic varicosities, many varicosities were not associated with detectable Sc processes. The present technique, and the capacity of confocal microscopy to yield three-dimensional imaging, allow the study of the microtopology of Sc and related sympathetic axons in the renal perivascular interstitiu

Immune response of healthy horses to DNA constructs formulated with a cationic lipid transfection reagent

Background Deoxyribonucleic acid (DNA) vaccines are used for experimental immunotherapy of equine melanoma. The injection of complexed linear DNA encoding interleukin (IL)-12/IL-18 induced partial tumour remission in a clinical study including 27 grey horses. To date, the detailed mechanism of the anti-tumour effect of this treatment is unknown. Results In the present study, the clinical and cellular responses of 24 healthy horses were monitored over 72 h after simultaneous intradermal and intramuscular application of equine IL-12/IL-18 DNA (complexed with a transfection reagent) or comparative substances (transfection reagent only, nonsense DNA, nonsense DNA depleted of CG). Although the strongest effect was observed in horses treated with expressing DNA, horses in all groups treated with DNA showed systemic responses. In these horses treated with DNA, rectal temperatures were elevated after treatment and serum amyloid A increased. Total leukocyte and neutrophil counts increased, while lymphocyte numbers decreased. The secretion of tumour necrosis factor alpha (TNFα) and interferon gamma (IFNγ) from peripheral mononuclear blood cells ex vivo increased after treatments with DNA, while IL-10 secretion decreased. Horses treated with DNA had significantly higher myeloid cell numbers and chemokine (C-X-C motif) ligand (CXCL)-10 expression in skin samples at the intradermal injection sites compared to horses treated with transfection reagent only, suggesting an inflammatory response to DNA treatment. In horses treated with expressing DNA, however, local CXCL-10 expression was highest and immunohistochemistry revealed more intradermal IL-12-positive cells when compared to the other treatment groups. In contrast to non-grey horses, grey horses showed fewer effects of DNA treatments on blood lymphocyte counts, TNFα secretion and myeloid cell infiltration in the dermis. Conclusion Treatment with complexed linear DNA constructs induced an inflammatory response independent of the coding sequence and of CG motif content. Expressing IL-12/IL-18 DNA locally induces expression of the downstream mediator CXCL-10. The grey horses included appeared to display an attenuated immune response to DNA treatment, although grey horses bearing melanoma responded to this treatment with moderate tumour remission in a preceding study. Whether the different immunological reactivity compared to other horses may contributes to the melanoma susceptibility of grey horses remains to be elucidated

Bacterium-like particles as multi-epitope delivery platform for Plasmodium berghei circumsporozoite protein induce complete protection against malaria in mice

Contains fulltext : 110364.pdf (publisher's version ) (Open Access)BACKGROUND: Virus-like particles have been regularly used as an antigen delivery system for a number of Plasmodium peptides or proteins. The present study reports the immunogenicity and protective efficacy of bacterium-like particles (BLPs) generated from Lactococcus lactis and loaded with Plasmodium berghei circumsporozoite protein (PbCSP) peptides. METHODS: A panel of BLP-PbCSP formulations differing in composition and quantity of B-cell, CD4+ and CD8+ T-cell epitopes of PbCSP were tested in BALB/c mice. RESULTS: BLP-PbCSP1 induced specific humoral responses but no IFN-gamma ELISPOT response, protecting 30-40% of the immunized mice. BLP-PbCSP2, with reduced length of the non-immunogenic part of the T-cell-epitopes construct, increased induction of IFN-gamma responses as well as protection up to 60-70%. Compared to controls, lower parasitaemia was observed in unprotected mice immunized with BLP-PbCSP1 or 2, suggestive for partial immunity. Finally, further increase of the number of B-cell epitopes and codon optimization (BLP-PbCSP4) induced the highest anti-CSP antibody levels and number of IFN-gamma spots, resulting in sterile immunity in 100% of the immunized mice. CONCLUSION: Presentation of Plasmodium-derived antigens using BLPs as a delivery system induced complete protection in a murine malaria model. Eventually, BLPs have the potential to be used as a novel versatile delivery platform in malaria vaccine development

Lipoplex-loaded microbubbles for gene delivery: A Trojan horse controlled by ultrasound

Cationic poly(ethylene glycol)ylated (PEGylated) liposomes are one of the most important gene transfer reagents in non-viral gene therapy. However, the low transfection efficiencies of highly PEGylated lipoplexes currently hamper their clinical use. Recently, ultrasound has been used in combination with microbubbles to enhance the uptake of genes in different cell types. However, the gene transfer efficiency still remains low in these experiments. To overcome the limitations of both techniques, we present the attachment of PEGylated lipoplexes to microbubbles via biotin-avidin-biotin linkages. Exposure of these lipoplex-loaded microbubbles to ultrasound results in the release of unaltered lipoplexes. Furthermore, these lipoplex-loaded microbubbles exhibit much higher transfection efficiencies than "free" PEGylated lipoplexes or naked plasmid DNA (pDNA) when combined with microbubbles and ultrasound. Interestingly, the lipoplex-loaded microbubbles only transfect cells when exposed to ultrasound, which is promising for space- and time-controlled gene transfer. Finally, this novel Trojan-horse-like concept can also be exploited to achieve the ultrasound- triggered release of nanoparticles containing other therapeutic agents such as anticancer drugs

Intranasal Delivery of Influenza Subunit Vaccine Formulated with GEM Particles as an Adjuvant

Nasal administration of influenza vaccine has the potential to facilitate influenza control and prevention. However, when administered intranasally (i.n.), commercially available inactivated vaccines only generate systemic and mucosal immune responses if strong adjuvants are used, which are often associated with safety problems. We describe the successful use of a safe adjuvant Gram-positive enhancer matrix (GEM) particles derived from the food-grade bacterium Lactococcus lactis for i.n. vaccination with subunit influenza vaccine in mice. It is shown that simple admixing of the vaccine with the GEM particles results in a strongly enhanced immune response. Already after one booster, the i.n. delivered GEM subunit vaccine resulted in hemagglutination inhibition titers in serum at a level equal to the conventional intramuscular (i.m.) route. Moreover, i.n. immunization with GEM subunit vaccine elicited superior mucosal and Th1 skewed immune responses compared to those induced by i.m. and i.n. administered subunit vaccine alone. In conclusion, GEM particles act as a potent adjuvant for i.n. influenza immunization