Memory foam är en bok om de obetydliga sakerna och platserna. Om minnen av stunder som egentligen inte betydde något och inte förändrade något, som inte fick några konsekvenser, inget hände, inget kom ut av det, men som av någon anledning ändå fastnat i mig. Saker som att vänta, ha tråkigt, åka bil, kvällar, andras hem, vänskaper där man gjorde inget särskilt. Jag samlar ihop fragmenten och lägger dem bredvid varandra. / Memory foam is a book about insignificant places and things. It is about memories of moments that did not really mean anything and that did not change anything. Moments that did not have any consequences, where nothing happened, nothing came from them, but for some reason stuck with me. Things like waiting, being bored, riding a car, evenings, other people’s homes, friendships where you did nothing special together. I collect the fragments and place them beside each other.

La thèse se propose d’étudier différentes formes de la représentation du passé dans le roman contemporain français. Si la recherche a pu constater, depuis un certain temps déjà, un véritable « retour de l’Histoire » sur la scène littéraire actuelle, force est de constater que chez certains auteurs, ce « retour » s’accompagne d’une méfiance profonde à l’égard de la connaissance de l’Histoire et des formes (culturelles, littéraires) de sa mise en récit. Ainsi Philippe Forest, dans Le Siècle des nuages (2010), raconte-t-il l’histoire de son père tout en se posant la question de savoir comment on peut écrire cette histoire qui constamment se dérobe. Il en sort un roman historique paradoxal et foncièrement autoréférentiel qui se construit et se déconstruit sous les yeux du lecteur. De façon comparable, Jean Rouaud, dans L’Imitation du bonheur (2006), invite son lecteur dans « l’atelier » de l’écrivain afin d’exhiber les sources et les ressources de son travail. Il ne se sert ainsi des techniques de l’illusion réaliste que pour mieux les parodier par la suite. Dans Waltenberg (2005) d’Hédi Kaddour, en revanche, la critique du récit historique se fait à travers la fragmentation de l’intrigue, la multiplication des temps et des lieux ainsi que par l’abandon de la linéarité. Aussi l’auteur nous présente-t-il une histoire éclatée qui se soustrait aux catégories d’unité et de cohérence. Bien que les stratégies de représentation dans ces trois romans soient diverses, elles renvoient pourtant à un concept commun que je propose ici d’appeler le récit historique critique.Dans la partie principale de mon travail, il s’agit d’analyser les trois romans mentionnés ci-dessus en tenant compte, notamment, des outils méthodologiques élaborés par la narratologie (D. Cohn, A. Nünning, W. Wolf) et par la théorie de l’histoire (H. White, P. Ricœur, R. Kosel-leck). Cette partie principale est précédée d’une partie plus « théorique » subdivisée en trois axes de recherche : L’approche systématisante essaie de condenser certaines des caractéristiques les plus importantes du récit historique critique dans une vue d’ensemble, élargissant par ailleurs le propos sur d’autres textes littéraires (contemporains). L’approche historicisante se propose de tisser un lien entre les romans analysés dans la partie principale de la thèse et l’œuvre de Claude Simon, considérée ici comme une sorte de paradigme esthétique pour une nouvelle écriture de l’histoire. L’approche comparative, enfin, tourne le regard vers les historiens professionnels (Ginzburg, Jablonka, Boucheron) qui, à l’instar des romanciers, ont exploré, ces derniers temps, de nouvelles formes de mise en récit du passé. Si ces nouvelles formes se rapprochent du récit littéraire par l’usage qu’elles font de la fiction, elles restent néanmoins sceptiques sur la possibilité même de représenter le passé, devenant de la sorte comme un équivalent en histoire du récit historique critique littéraire. / This PhD thesis offers a study of different forms of representations of the past in the French contemporary novel. Since several years, literary critics agree that there has been something like a “return to history” on the actual literary scene. But we also have to state that some con-temporary authors combine this “return” with a profound suspicion against the knowing of history and the (cultural or literary) forms of telling it. In his Le Siècle des nuages (2010), Philippe Forest tells the story of his father while at the same time questioning himself about how to represent a story which seems to withdraw itself constantly. The result is somewhat paradoxical and may be described as a self-referential historical novel that constructs and deconstructs itself in front of the reader’s eyes. In a similar way, Jean Rouaud’s L’Imitation du bonheur (2006) invites its reader in the artist’s workshop in order to reveal the sources and the resources of the story. Exploiting the techniques of realistic illusion, Rouaud is in fact parodying them. Waltenberg (2005) by Hédi Kaddour acts differently: in this novel, historical narration is undermined by a broken storyline, by a pluralization of times and spaces and by the abandoning of the chronological order. Thus the author confronts us with a history “in pieces” that doesn’t obey the principles of unity and coherence anymore. Even if the strategies of historical representation are quite different in these three novels, they are all based on a common concept which I suggest to call “critical historical narration”.The main part of my work consists of the analysis of the three novels mentioned above, an analysis which is mostly based on methods provided by narratology (D. Cohn, A. Nünning, W. Wolf) or theory of history (H. White, P. Ricœur, R. Koselleck). This main part is preceded by a more general part that is subdivided into three theoretical approaches: the systematizing approach tries to reunite some of the most important features of critical historical narration in a synopsis and thereby takes also account of other (contemporary) literary texts. The historical approach connects the novels analysed in the main part of this thesis to the work of Claude Simon, which will be considered here as an esthetical paradigm for a new kind of historical narration. Finally, the comparative approach focuses on professional historians (Ginzburg, Ja-blonka, Boucheron) who have been exploring new strategies of historical writing in the same way that novelists have done. Indeed, if these new strategies converge with those in literary writing because of the use they make of fiction, they are nevertheless included in a more general reflection on the possibilities of representing the past. Historiography thus may provide something like an equivalent to critical historical narration in literature.

L’écriture fragmentaire est le lieu d’un flou théorique. Elle est tout d’abord le fruit d’une appréhension intuitive, pragmatique : est fragmentaire tout texte perçu comme tel. D’Héraclite aux surréalistes en passant par Montaigne ou les frères Schlegel, l’écriture fragmentaire est protéiforme. Cependant, dans le champ de la théorie littéraire française, elle n’est généralement envisagée que dans son versant aphoristique et définie à partir d’un corpus composé de textes au statut littéraire à la limite d’un autre champ disciplinaire (la critique, la philosophie), majoritairement en langues occidentales, relevant d’une énonciation sérieuse et factuelle dont la fragmentation relève d’une démarche consciente de pensée et d’écriture. Cette thèse a ainsi pour objectif de montrer, à travers le cas particulier de la poétique singulière du fragment chez Akutagawa Ryūnosuke, l’existence d’un fragment non purement factuel, provenant d’une aire culturelle non occidentale, qui s’inscrit plus largement dans cette écriture de la crise qu’est le fragment moderne selon Françoise Susini-Anastopoulos. Les textes d’Akutagawa qui font l’objet de cette étude sont de nature variée : narratifs, autobiographiques, factuels, fictionnels, aphoristiques ou encore poétiques, ils relèvent néanmoins d’une même esthétique fragmentaire. En confrontant ces textes d’un écrivain japonais du début du XXe siècle aux réflexions théoriques tant japonaises que françaises sur les écritures brèves discontinues, nous tentons de redéfinir les contours d’une écriture fragmentaire littéraire tout en proposant une nouvelle grille de lecture pour des textes divers pour lesquels la catégorisation générique est souvent problématique. Le fragment chez Akutagawa s’articulerait ainsi autour de deux pôles : un brouillage du cadre générique dans lequel s’inscrit le texte, ainsi qu’une énonciation ironique, souvent secondée par un usage prégnant de l’intertextualité. / Fragmentary writing is difficult to define. It is, first of all, the result of a pragmatic, intuitive understanding: a text is called fragmentary when it is perceived as such. From Heraclitus, Montaigne, the Schlegel brothers to the surrealists, fragmentary writing takes many forms. However, in French literary theory, it is often limited to aphoristic texts. Its corpus is made of literary texts mostly written in European languages, which could also belong to another field (like critic or philosophy), which are non-fictional, and where fragmentation is a conscious, voluntary process of thinking and writing.This thesis aims to show, through the concrete example of Akutagawa Ryūnosuke’s poetics of fragmentary writing, the existence of a fragment outside the fiction / non-fiction dichotomy, from a non-western cultural area, which belongs to the modern fragment as crisis literature defined by Françoise Susini-Anastopoulos.In this thesis, we look at various texts written by Akutagawa. Be they narrative, fictional, non-fictional, autobiographical, poetic or aphoristic: all these texts have a common fragmentary aesthetic, despite their diversity. By reading these texts written by a Japanese writer who lived at the beginning of the 20th century under the light of Japanese and French critical works on short and discontinuous writings, we are trying to redefine the outline of a literary fragmentary writing while suggesting a new way of reading them beyond the very different generic categories they are usually thought to belong to. Accordingly, two features could describe Akutagawa’s fragmentary writing: a problematical generic categorization deliberately constructed in the texts, and an ironic voice, which is often accompanied by intertextuality. This thesis aims to show, through the concrete example of Akutagawa Ryūnosuke’s poetics of fragmentary writing, the existence of a fragment outside of the fiction / non-fiction dichotomy, from a non-western cultural area, which belongs to the modern fragment as crisis literature defined by Françoise Susini-Anastopoulos.In this thesis, we look at various texts written by Akutagawa. Narrative, fictional, non-fictional, autobiographical, poetic, aphoristic: all the texts have, despite their diversity, a common fragmentary aesthetic. By reading these texts, written by a Japanese writer who lived at the beginning of the 20th century, under the light of Japanese and French critical works on short and discontinuous writings, we are trying to redefine the outline of a literary fragmentary writing while suggesting a new way of reading problematical texts in terms of literary genre. Accordingly, two features could describe Akutagawa’s fragmentary writing: a problematical generic categorization deliberately constructed in the text and an ironic voice, which is often accompanied by a prominent intertextuality.

Tuberculosis affects millions of people worldwide every year. The current treatment for TB is divided into a regimen of both first- and second-line drugs, where first-line treatments are more tolerated and require shorter treatment lengths. With rising levels of resistance, alternative treatment regimes are urgently needed to fight this disease. Ethionamide, a second-line drug is administered as a prodrug which is activated in vivo by the enzyme EthA, which is in turn regulated by EthR. The disruption of the action of EthR could lead to novel therapeutics which could enhance the efficacy of ethionamide, and raise it to a first-line treatment. The work reported in this thesis examines the elaboration of three chemical scaffolds using fragment-based approaches to develop novel inhibitors capable of disrupting the EthR-DNA interaction. The first scaffold, 5-(furan-2-yl)isoxazole was investigated by fragment-merging approaches and produced compounds with the best of these having a KD of 7.4 uM. The second scaffold, an aryl sulfone was elaborated using fragment-merging strategies. This led to several modifications of the fragment, leading to several variants with KDs around 20 uM. With both of these series the affinity could not be improved below 10 uM and due to the synthetic complexity a further scaffold was prioritised. The third scaffold was explored was a 4-(4-(trifluoromethyl)phenyl)piperazine using fragmentgrowing from the NH of the piperazine to probe deeper into the EthR binding pocket. In addition to this, SAR around the 4-(trifluoromethyl)phenyl group was assessed to explore the interactions with EthR. These modifications led to compounds with nanomolar IC50s. A range of compounds were then screened by REMAssay to determine the boosting effect on ethionamide, and this identified compounds with up to 30 times boosting in the ethionamide MIC. The final chapter examines a concept where compounds were designed to exploit the dimeric nature of EthR by linking two chemical warheads with a flexible linker. These compounds are examined using mass spectrometry to investigate the stoichiometry of the interaction to provide insight into the binding of these extended compounds and exploring an alternative strategy to inhibit EthR. The work in this thesis demonstrated the successful use of fragment-based approaches for development of novel EthR inhibitors which showed significant ethionamide boosting effects.

Ce travail de recherche contribue au questionnement sur les nouveaux enjeux de conception soulevés par le design critique. À plusieurs égards, le corpus de cette thèse repose sur une histoire de la contre-histoire. Une manière de faire et de penser dont l’historicité la plus évidente se situe à la fin des années 1960, lorsque certains designers ont souhaité porter le projet vers la mise en œuvre d’un écart afin d’interroger la société par les moyens du design. Une attitude qui instaure la conception à l’instar d’une logique de la désadaptation des habitudes et des manières conventionnelles de penser. La notion de critique, du grec krinein, qui signifie «séparer», «distinguer», prend ici tout son sens en instaurant, non pas une rupture, mais un déplacement, qui aménage des entre-deux et ménage des porosités de la pensée et de la conception. De l’individu augmenté à l’individu modifié, cette thèse révèle de surcroît, l’inquiétude d’une mainmise au delà de l’invention de l’artificiel et aux abords du biologique. Au gré d’un épuisement de la question du critique en design, nous décelons des moyens de déconstruire les systèmes de la discipline et ses réinventions. / This research work aims to contribute to the study of the new challenges of conception raised by critical design. In several respects the corpus of this thesis is based on a history of coun¬ter history, a way of doing and thinking whose conspicuous historicity takes place in the late sixties when a few designers decided to reconsider the notion of project and stray from the straight and narrow in order to question society through the means of design. The notion of criticism - from the Greek krinein meaning «separate», «discriminate» - takes on its full meaning by introducing a shift (not a break) which makes room for inter¬vals and allows sensitive thinking and conception. From human enhancement to human modification, this thesis also reveals a number of concerns which go beyond the invention of artificial means and touch upon biological aspects. Considering that the critical dimen¬sion in the field of design has been exhausted, this research highlights a few ways to deconstruct the operating principles of this discipline as well as its reinventions.

Transmissive spongiform encephalopathies (TSEs) are neurodegenerative diseases characterized by depositions of abnormally folded prion protein (PrPTSE ) in brain. PrPTSE is at present the only specific biochemical marker of human and animal TSEs. Diagnostic tests are based on the detection of PrPres after proteinase K digestion of brain homogenate using Western blot or on the immunohistochemistry of fixed brain tissue, which are both difficult and time consuming. In this work we focused on development of a new type of tests based on PrP detection without need of proteinase K digestion. As deposits of PrPTSE remain in the body for a long time, there is a substantial chance of them being nonenzymatically modified by glycation. The detection of glycated PrPTSE may have a potential to serve as a diagnostic marker. We prepared monoclonal antibodies specific for carboxymethyl lysine/arginine modified prion protein. Bacterially expressed and purified recombinant human prion protein (rhPrP) was modified by glyoxylic acid that introduces carboxymethyl groups on lysine and arginine residues present within the molecule of the protein. Modified rhPrP (rhPrP-CML) was used for immunization of laboratory mice and hybridoma cells were prepared. Screening of cell supernatants resulted in the selection of 4...

Influenza virus is a pleiomorphic human pathogen which causes self-limiting respiratory illness lasting one-two weeks in most individuals. However, in immunologically compromised individuals, influenza infection may lead to severe morbidity and fatality. Annual epidemics cause 250,000-500,000 deaths worldwide and remain a major public health threat. The virus has evolved mechanisms of antigenic ‘drift’ and ‘shift’ to evade the host immune response. Hence, current influenza vaccines need to be updated every few years. Moreover, the currently available inactivated/live attenuated vaccines entail virus culture in embryonated chicken eggs hindering rapid scale-up. The aforementioned limitations of the current vaccines has had debilitating effect when strain mismatch between vaccine formulation and influenza viruses circulating within the population has occurred in the past, despite intensive monitoring. Public health is further compromised when an unpredictable mixing event among influenza virus genomes leads to antigenic shift, facilitating a potential pandemic outbreak. These concerns have expedited efforts towards developing a ‘universal’ flu vaccine. Influenza hemagglutinin (HA) is the primary target of the humoral response during infection/vaccination. The precursor polypeptide, HA0, is assembled into a trimer along the secretory pathway and transported to the cell surface. Cleavage of HA0 generates the mature, disulfide linked HA1 and HA2 subunits. Mature HA has a globular head domain which mediates receptor binding and is primarily composed of the HA1 subunit while the stem domain predominantly comprises of the HA2 subunit. The HA stem is trapped in a metastable state and undergoes an extensive low-pH induced conformational rearrangement in the host-cell endosomes to adopt the virus-host membrane fusion competent state. The ‘antigenic sites’ on the immunodominant globular head of HA are subjected to heightened immune pressure resulting in escape variants, thereby limiting the breadth of head-directed neutralizing antibodies (nAbs). As opposed to the highly-variable head domain, the HA stem is conserved and targeted by several broadly neutralizing antibodies (bnAbs) with neutralizing activity against diverse influenza A virus subtypes. Although several bnAbs bind to the conserved HA stem, focusing the immune response to this conserved, subdominant stem domain in presence of the variable head domain of HA has been challenging. Alternatively, mimicking the epitope of these stem-directed bnAbs in the native, pre-fusion conformation in a ‘headless’ stem immunogen capable of eliciting a broadly protective immune response has been difficult because of the metastable conformation of HA. Addressing the aforementioned challenges, we describe the design and characterization of novel influenza immunogens by HA protein minimization. Chapter 1 gives an overview of the influenza virus life cycle, and outlines the structural organization and function of viral proteins. The conventional vaccines that are currently used and their limitations are described in this chapter. Recent improvements in influenza vaccine production focusing on recombinant HA as an alternate solution are discussed. Painstaking efforts of several groups in the recent past has led to the isolation of bnAbs that recognize novel ‘antigenic signatures’ within the globular head and the HA stem domains. Attempts to focus the immune response to these ‘cross-protective’ epitopes are described. The design and characterization of trimeric HA stem-fragment immunogens from influenza A Group-1 viruses which mimic the native, pre-fusion conformation of HA are described in Chapter 2. We engineered ‘headless’ HA stem immunogens based on influenza A/Puerto Rico/8/34 (H1N1) subtype. H1HA10-Foldon, a trimeric derivative of our parent construct (H1HA10), bound conformation sensitive stem-directed bnAbs such as CR6261, F10 and FI6v3 with high affinity (equilibrium dissociation constant [KD] of 10-50nM). The designed immunogens elicited broadly cross-reactive antiviral antibodies which neutralized highly drifted influenza virus strains belonging to both Group-1 (H1, H5 subtypes) and 2 (H3 subtype) in vitro. Significantly, stem immunogens designed from unmatched, highly drifted influenza strains conferred protection against a lethal (2LD90) heterologous A/Puerto Rico/8/34 virus challenge in mice. Our immunogens conferred robust subtype-specific and modest heterosubtypic protection in vivo. In contrast to previous HA stem domain immunogens, the designed immunogens described here were purified from the soluble fraction in E.coli. These HA stem-fragment immunogens do not aggregate even at high concentrations and are cysteine-free which eliminates the complications arising from incorrect disulfide-linked, misfolded conformations. The aforementioned properties of the HA stem-fragment immunogens make it amenable for scalability at short notice which is vital during pandemic outbreaks. The detailed mechanism(s) by which our ‘headless’ stem immunogens provide protection need further investigation. The long central α-helices (LAH) located in the HA stem assemble together into a parallel, trimeric coiled-coil. Immunization with the wt-LAH (76-130 of HA2) derived synthetic peptide designed from an H3 subtype (H3N2 A/Hong Kong/1/68) and conjugated to keyhole limpet hemocyanin (KLH) was shown previously to elicit antibodies reactive in ELISA with multiple hemagglutinin subtypes and to confer protection against challenge with H3N2, H1N1 and H5N1 virus strains. The LAH peptide sequence was chosen based on maximal binding to the monoclonal antibody (MAb), 12D1, which has broad neutralizing activity against influenza viruses of the H3 subtype. These results motivated us to rationally design stabilized derivatives of wt-LAH and test their protective capacity in a mouse challenge model of influenza. This work is described in Chapter 3. Additionally, to understand the contribution towards protection conferred by the two distinct surface exposed patches on LAH, we designed constructs spanning different stretches of LAH. The biophysical characterization of the LAH-derived constructs indicates that most of them were well-folded. All these constructs were moderately immunogenic in mice but at best, conferred limited protection from lethal viral challenge. In contrast to previously reported results, our data suggests that the LAH in the absence of other regions of HA may require not only strong, but also specific adjuvantation to induce a robust and functional immune response in vivo. Chapter 4 describes an immunogen design (H1pHA9) based on the globular head domain of pandemic H1N1 HA which can be produced using a prokaryotic expression system. The HA-fragment, H1pHA9, stably refolds to mimic the conformation sensitive neutralizing epitopes in the globular head domain of HA. We have also successfully engineered the HA head domain to delineate the epitope of antibodies neutralizing the pandemic H1N1 virus using a yeast cell-surface display platform. In this direction, we report the isolation of a novel, neutralizing murine MAb, MA2077, against the pandemic H1N1 virus. The epitope of this MAb has been mapped onto the ‘Sa’ antigenic site. The ability of the head domain fragment, H1pHA9, which binds MA2077 with high affinity to elicit such neutralizing antibodies in vivo needs to be further explored. Structural analysis has shown that elements of the HA stem diverge between the two phylogenetic groups. Therefore, to mitigate the threat of circulating influenza A viruses from these distinct structural classes (H1 and H3 belonging to Groups 1 and 2 respectively), in lieu of a ‘universal’ vaccine, a combination of immunogens derived from both the groups is a practical alternative. In Chapter 5 we describe the design of stem-fragment immunogens from an influenza A Group-2 virus strain. We report the characterization of engineered ‘headless’ HA stem immunogens based on influenza A/Hong Kong/1/68 (H3N2) subtype. The designed immunogens were expressed in E.coli and purified from the soluble fraction with abundant yields (~15mg/lt). The HA stem-fragment immunogens could be concentrated to high concentrations without aggregation. While, H3HA10-IZ and H3HA10-Foldon, the trimeric derivatives of our parent construct (H3HA10) which were folded, conferred modest protection against a lethal homologous virus challenge in mice, there is considerable scope to improve our immunogen design. Analyzing the results from our previous work (Chapter 2), we speculate that structural elements at the N-terminus of A-helix are critical for helix initiation. We therefore extended the design to include residues from the start of the A-helix. We designed the extended stem immunogens from both H3 and H7 subtypes. The proteins were purified from the soluble fraction of the E.coli cell culture lysate. Preliminary studies suggest that extension of the A-helix has aided proper folding. These proteins need to be further characterized and evaluated in an animal model.

The goal of this work is to create an application, which will combine procedural and keyfram animations with subsequent visualization. Composition of this two different animations techniques is used to animate a virtual character. To combine this two techniques one starts with interpolations from keyframe animation and then enchance them by procedural animations to properly fit into the characters surroundings. This procedural part of animation is obtained by using forward and inverse kinematics. Whole application is written in C++, uses GLM math library for computations and OpenGL and GLUT for final visualization.

The first part of this project is focused on theoretical analysis of methods for rendering volume data. Both methods are analyzed showing the volume data using triangle mesh, and methods for direct volume rendering. Ray Casting is presented in detail. Possible way of its realization using graphics card is the subject of implementation part. The paper presents several methods that could be applied to ray casting and achieve different results of visualization of the same data. The work also aims to create a  graphical user interface that allows interactive visualizations.

L’ischémie/reperfusion (I/R) occurrente à toutes transplantations d’organe solide, constitue un stimulus pro-autophagique/pro-apoptotique pour les cellules endothéliales. Nous avons récemment démontré que les cellules endothéliales apoptotiques (CEapo) sécrètent des vésicules apoptotiques exosome like (ApoExo) qui induisent une réponse auto-immune et accélèrent le rejet vasculaire dans un modèle de greffe aortique chez la souris. Ces ApoExo qui diffèrent des corps apoptotiques classiques par leur structure et leur contenu protéique contiennent le fragment C-terminal du perlécan, LG3. Le LG3, un auto-antigène d’importance en transplantation, favorise le remodelage vasculaire et est augmenté en circulation chez les patients greffés rénaux atteints d’un rejet vasculaire. De plus, la présence d’anticorps anti-LG3 avant la transplantation est associée à un risque plus élevé de développer un rejet vasculaire chez des patients greffés du rein et à la perte du greffon à long terme. Il est connu que la génération du fragment LG3 implique la protéolyse du perlécan par la cathepsine L, une protéase lysosomiale, mais le mécanisme de transport de ce fragment au sein des ApoExo est encore inconnu. Nous avons émis l’hypothèse que l’activité lysosomiale et l’autophagie jouent un rôle important dans la maturation et le chargement du LG3 dans les vésicules ApoExo sécrétées par les CEapo. Une étude longitudinale de microscopie électronique chez les cellules endothéliales humaines carencées en sérum pendant 1h, 2h et 3h, nous a révélé la présence du perlécan / fragment LG3 au sein de compartiments autophagiques (autophagosomes et autophagolysosomes) à différentes étapes du processus autophagique. Après 3 heures de carence en sérum, nous avons identifié le fragment LG3 dans des vésicules membranaires situées au sein de grands réseaux vacuolaires s’apparentant à des autophagolysosomes. L’inhibition de la cathepsine L, de l’acidification du lysosome et de l’autophagie diminuent la présence du fragment LG3 dans les vésicules ApoExo sans affecter la sécrétion de vésicules démontrant donc le rôle de l’autophagie dans l’intégration du fragment LG3 au sein des ApoExo. Toutefois, l’injection de vésicules ApoExo LG3-, provenant de cellules murines aortiques traitées à la bafilomycine, dans un modèle murin de greffe aortique induit une réponse auto-immune anti-LG3 ainsi qu’un remodelage vasculaire à des niveaux similaires aux souris ayant reçu des ApoExo véhicule. Une analyse protéomique de ces vésicules ApoExo LG3-, nous a révélé que la bafilomycine modifie le contenu protéique des ApoExo en induisant une augmentation de la présence de protéines lysosomiales et de la matrice extracellulaire dont le perlécan. Ceci suggère que la présence du motif LG3 au sein du perlécan natif non clivé dans les ApoExo LG3- pourrait être responsable de la mise en place de la réponse anti-LG3 observées chez les souris. Les travaux de ce doctorat ont permis de mettre en évidence que l’autophagie dans les cellules endothéliales productrices d’ApoExo ne régule pas l’immunogénicité des ApoExo. Cependant, elle régule au sein des cellules endothéliales apoptotiques le transport et le clivage du perlécan qui lui est immunogène. En effet, l’autophagie module les différentes formes de perlécan natif et clivé sécrétées dans les ApoExo. L’étude chez la souris greffée nous a donc permis de considérer non plus l’implication du fragment LG3 mais l’implication du motif LG3 présent au sein du perlécan natif non clivé et de ses différentes formes intermédiaires dans la réponse auto-immune anti-LG3 et le rejet vasculaire. La modulation de la sécrétion du perlécan et du LG3 dans les ApoExo constitue une cible thérapeutique potentielle afin de diminuer la réponse auto-immune pouvant augmenter le dommage vasculaire après la greffe / Ischemia/reperfusion (I/R) occurring in all solid organ transplantation, constitute a proautophagic / pro-apoptotic stimulus on endothelial cells. We recently demonstrated that apoptotic endothelial cells (CEapo) secrete apoptotic exosome-like vesicles (ApoExo) that induce autoimmune response and accelerate vascular rejection in a mouse aortic transplant model. These ApoExo, that differ from classical apoptotic bodies in structure and protein content, contain the C-terminal fragment of perlecan, LG3. LG3, an autoantigen of importance in transplantation, promotes vascular remodeling and is increased in circulation in renal transplant patients undergoing vascular rejection. In addition, the presence of anti-LG3 antibodies prior to transplantation is associated with a higher risk of developing vascular rejection in kidney transplant patients and long-term graft loss. It is known that the generation of LG3 fragment involves the proteolysis of perlecan by cathepsin-L, a lysosomal protease, but the mechanism of export of this fragment within ApoExo is still unknown. We hypothesized that lysosomal activity and autophagy play an important role in the maturation and the secretion of LG3 in ApoExo vesicles secreted by CEapo. Longitudinal electron microscopy study after 1h, 2h and 3h in serum starved endothelial human cells revealed the presence of perlecan / LG3 fragment within autophagic compartments (autophagosomes and autophagolysosomes) at different stages of the autophagic process. After 3 hours of serum starvation, we identified LG3 fragment in membrane vesicles located within large vacuolar networks reminiscent of autophagolysosomes. Inhibition of cathepsin-L, of lysosomal acidification and of autophagy decrease the presence of LG3 fragment in ApoExo vesicles without affecting vesicle secretion thus demonstrating the role of autophagy in the secretion of LG3 fragment within ApoExo. However, Injection of ApoExo LG3- vesicles from bafilomycin-treated aortic murine cells into a murine aortic transplant model induces autoimmune anti-LG3 response and vascular remodeling at levels similar to the ApoExo vehicle mice control group. Proteomic analysis of ApoExo from bafilomycin-treated aortic murine cells has demonstrated that bafilomycin modifies ApoExo protein content by inducing an increase of the presence of lysosomal proteins and the extracellular matrix, including perlecan. This suggests that the presence of LG3 motif in uncleaved native perlecan in ApoExo LG3- could be responsible for the establishment of the anti-LG3 response as well as the vascular remodeling observed in mice. Collectively, these results demonstrate that autophagy in endothelial cells producing ApoExo does not regulate the immuogenicity of ApoExo. However, it regulates within apoptotic endothelial cells the processing and cleavage of perlecan who is immunogenic. Indeed, autophagy modulates the different forms of native and cleaved perlecan secreted in ApoExo. Grafted mice study thus allowed us to consider neither the involvement of the LG3 fragment but the implication of the LG3 motif present in the uncleaved native perlecan and its intermediate forms in the anti-LG3 autoimmune response and vascular rejection. Modulating perlecan and LG3 secretion in ApoExo vesicles is a potential therapeutic target to reduce the autoimmune response that can increase vascular damage after transplantation.