Un paradigme actuel en virologie est que les virus de plantes ne peuvent pas affecter les animaux vertébrés et ne sont pas pathogènes pour l'homme. Cependant, plusieurs éléments remettent en question ce paradigme. L'objectif de cette Thèse a été d'étudier si les tobamovirus peuvent être pathogènes chez l'homme. Ce manuscrit comprend :1) une revue de la littérature portant sur la présence et l'impact potentiel des virus de plantes chez les animaux dont l'homme. 2) une étude sur l'exposition humaine aux tobamovirus et les effets possibles Dans la région de Marseille, nous avons montré la présence du PMMoV dans 7.2% des selles de patients et dans 57 % des produits alimentaires à base de piment. De plus, la présence du PMMoV dans les selles de patients a pu être corrélée des signes cliniques. De plus, nous avons montré que le TMV infectieux était présent dans le tabac de toutes les cigarettes testées mais aussi dans 45 % des salives de fumeurs testées. 3) une étude in vivo. Suite à une inoculation intra-trachéal des souris avec du TMV, nous avons constaté que le virus persiste jusqu'à 14 jours dans le tissu pulmonaire et les macrophages pulmonaires. De plus, nous avons détecté du TMV dans le cytoplasme des macrophages murins jusqu'à 15 jours après inoculation. 4) une étude sur le mode de transmission du TMV à l'homme. Le TMV détecté dans la salive des fumeurs ne serait pas véhiculé par la fumée de cigarette, mais par contact avec du tabac infecté. Nos résultats suggèrent que la frontière entre virus de plantes et virus d'animaux n'est pas aussi stricte qu'il est communément admis et incitent à réévaluer l'éventuelle pathogénicité des phytovirus pour l'homme. / A current paradigm in virology is that plant viruses cannot affect vertebrates and are not pathogenic for humans. However, several recent findings challenge this paradigm. The aim of this thesis was to investigate whether tobamoviruses may be pathogenic in humans. This manuscript contains: 1) A review of the literature of the reasons why plant viruses may cross the border from plants to vertebrates and the possible impact of plant viruses in animals including humans. 2) A study on exposure to tobamovirus and possible effects of these viruses for humans. In the Marseille geographical area, we showed the presence PMMoV in 7.2% of stools from patients tested and in 57% of food products containing hot peppers. In addition, the presence of PMMoV in the stools of patients could be significantly correlated with clinical symptoms. Then, we showed that infectious TMV was present in the tobacco of all cigarettes tested and TMV RNA was detected in 45% of smokers' saliva tested. 3) In vivo study: mice intra-tracheal inoculation with TMV. The results showed the persistence until 14 days of viruses in the lung tissue and in lung alveolar macrophages of mice. In addition, we detected TMV in the cytoplasm of murine macrophages up to 15 days after inoculation. 4) A study of TMV transmission to humans through cigarette smoking. TMV does not seem to be vehicled by cigarette smoke to smoker's saliva but by direct contact with infected tobacco. Our results suggest that the boundary between plant viruses and animal viruses is not as strict as it is commonly accepted and prompt to reassess the potential pathogenicity of these plant viruses for humans.

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Les possibilités de combinaison thérapeutiques offertes par les nanoparticules ont ouvert un nouveau champ d’investigation pour la recherche sur le cancer. Dans ce projet de recherche, des nanoparticules dérivées de la protéine de capside du virus de la mosaïque du tabac (TMV) ont été utilisées afin de transporter différents peptides thérapeutiques ciblant le récepteur neuropiline-1. Cette stratégie a permis de solubiliser un peptide fortement hydrophobe ayant préalablement démontré son efficacité anticancéreuse sur des lignées de cancer du sein humain et de glioblastome. Les résultats obtenus ont également permis de démontrer la possibilité de combiner différents peptides thérapeutiques via l’auto-assemblage de la protéine de capside du TMV. / Nanoparticles play an ever increase role in carrying therapeutic compounds in the cancer field. In this research project, the coat protein of Tobacco mosaic virus (TMV) was used as nanocarrier to solubilize a hydrophobic peptide interfering with the transmembrane domain of neuropilin-1. The nanoparticles created have conserved the antiangiogenic and antimigratory effect of the therapeutic peptide. This strategy was also used to create nanoparticles carrying a peptide targeting the ectodomain of neuropilin-1. The two types of nanoparticles were then assembled through auto-assembling ability of the coat protein. These nanoparticles also exhibit antiangiogenic ability thus, confirming the validity of this approach to combine therapeutic peptides.

Le VRP permet de modéliser la problématique du TMV. Pour autant dans sa version la plus simpliste il ne peut convenir aux nouvelles contraintes qui renforcent la complexité de l’environnement urbain tel que la congestion. Raison pour laquelle ce travail se positionne en premier sur un TDVRP (Time-Dependent-Vehicle-Routing-Problem) dans le but de représenter au mieux le contexte urbain actuel. La résolution de ces deux problèmes est efficace, entraînant une réduction du nombre de véhicules supplémentaires nécessaires en raison des retards de congestion peuvent être éliminés. Mais cette solution n’est pas sans conséquence pour les clients finaux puisque son but est d’éviter le passage aux heures de pointes et cela peut engendrer des retards de livraison. Pour cette raison nous voulons proposer une nouvelle solution qui permet de résoudre le problème du chemin le plus court et en intégrant la dépendance au temps (TDVRP). Cette solution se repose sur une meilleure exploitation des véhicules, en premier lieu, pour réduire le nombre de véhicule sur la route. Ce problème s’appelle MT-VRP (Multi-Trip VRP) qui a été évoqué en 2007 par (Azi, et al. 2007). Dans un deuxième temps nous voulons proposer un MT VRP qui utilise différents types de véhicule en termes de capacité de chargement. Dans cette optique vient notre travail qui propose une nouvelle solution de MTTDVRP (TW avec fenêtres de temps) en testant une nouvelle solution en utilisant un véhicule léger de faible capacité de chargement (type tricycle) dans un but de réduire les nuisances liés au TMV. / The VRP makes it possible to model the problem of TMV. However, in its most simplistic version it cannot be adapted to the new constraints which reinforce the complexity of the urban environment such as congestion. This is why this work is placed first on a TDVRP (Time-dependent-Vehicle-Routing-Problem) in order to best represent the current urban context. The resolution of these two problems is effective; resulting in a reduction in the number of additional vehicles needed due to congestion delays can be eliminated. But this solution is not without consequences for end customers since its goal is to avoid the switch to peak hours and this can lead to delivery delays. For this reason we want to propose a new solution that solves the problem of the shortest path and integrating time dependency (TDVRP). This solution relies on better operation of vehicles, in the first place, to reduce the number of vehicles on the road. This problem is called MT-VRP (Multi-Trip VRP) which was raised in 2007 by (Azi, et al. 2007). In a second step we want to propose a MT VRP that uses different types of vehicle in terms of loading capacity. In this perspective comes our work that proposes a new solution of MTTDVRP (TW with time windows) by testing a new solution using a light vehicle of low load capacity (tricycle type) in order to reduce the nuisance associated with TMV.

Infectious diseases remain a significant cause of human deaths, as approximately 15 million deaths were attributed to infectious diseases in 2010 (Dye, 2014). One such disease is rabies, which causes around 59,000 human deaths worldwide annually according to some estimates (Kessels et al., 2017). However, 95% of human deaths attributed to rabies occur in Asia and Africa (Singh et al., 2017). Rabies is preventable, yet it is still a major concern in developing, low-income countries that lack access to the medical care necessary to combat it (Hampson et al., 2015). Alternative techniques for low-cost vaccine production have the potential to resolve this issue. This research investigates the use of recombinant DNA techniques and plant biotechnology to produce a more cost-effective vaccine for rabies. Gene sequences from the rabies glycoprotein were inserted at the end of the coat protein portion of the Tobacco Mosaic Virus (TMV) genome. Plants were then infected with this recombinant virus, with hopes that TMV particles would assemble with proteins produced from the inserted glycoprotein sequence fused to the TMV coat protein. Results thus far suggest some of the sequences could be producing recombinant TMV particles, although issues involving successful extraction and reversion to wild type are still a challenge. Additionally, other research suggests that this is an effective method for vaccine development in general and for rabies.

Le transport de marchandises en ville (TMV) est essentiel à la vitalité économique des espaces urbains, mais également source de nombreuses externalités négatives, notamment la congestion routière. Si la quantification des pertes de temps est aujourd’hui une pratique banalisée, la contribution du TMV est moins fréquemment étudiée. Par ailleurs, l’impact réciproque de la congestion sur les opérations de TMV est relativement peu abordé, surtout dans une perspective urbanistique et géographique. L’occupation de la voirie par les véhicules de fret et leurs impacts sur l’organisation du TMV peuvent pourtant être considérés comme étant le résultat de divers phénomènes socio-spatiaux.Cette thèse se propose d’analyser l’occupation de la voirie par les véhicules de fret, l’importance des coûts de congestion routière associés ainsi que les incidences sur l’organisation du TMV. D’un point de vue théorique, le TMV est pensé comme un ensemble de processus industriels (à envisager dans un cadre plus large que la simple opération de transport) aux caractéristiques variées, mis en place par des entreprises pour répondre aux besoins d’approvisionnement des entreprises d’une aire urbaine. Nous analysons l’articulation de ces processus industriels avec les formes urbaines de la ville, et en particulier la relation difficile qu’entretiennent le TMV et la densité spatiale. D’un point de vue empirique, nous mobilisons plusieurs sources de données décrivant les flux de marchandises en Ile-de-France (dont l’enquête Transport de Marchandises en Ville (ETMV) réalisée en 2010). Outre son ancrage fort dans un référentiel spatial, nos analyses empiriques de la contribution du TMV à la congestion routière s’inspirent de plusieurs cadres disciplinaires, l’ingénierie des trafics et l’économie des transports notamment / Urban goods movements (UGM) are essential for economic activities in urban areas, but they are also generators of negative externalities, including road congestion. However, while the measure of time losses is a common research practice, the contribution of UGM is less frequently studied. The reciprocal impact of congestion on UGM operations is also rarely addressed, especially from the perspective of urban planning and geographic analysis. Yet, road occupancy by freight vehicles, and their impacts on UGM operations and organization can be considered as the results of socio-spatial phenomena.This thesis proposes an analysis of road occupancy by freight vehicles, of the induced congestion costs, and of the impacts on UGM operations. From a theoretical perspective, we analyze UGM as various industrial processes (which should be considered in a wider perspective than the simple transport operation), with diverse characteristics, implemented by companies to meet the supply needs of companies of an urban area. We also emphasize the need to analyze the links between those industrial processes and urban forms, and we focus on the difficult relationship between UGM and urban density. From an empirical perspective we rely on several data sources (including the Urban Goods Movements Survey (UGMS) performed in Ile-de-France in 2010). While our framework is grounded in spatial analysis, our empirical analyses of road congestion induced by urban freight also borrow from several disciplinary fields, including traffic engineering and transport economics

Tobacco mosaic virus (TMV) is one of the most investigated viruses and its attributes and structure are therefore well-known. In this work, we have chosen TMV as a biotemplate for the adjustable-length particles production in plants. The viral RNA and coat protein of TMV self-assemble into particles under physiological conditions. The particle length depends on the length of packaged RNA. The encapsidation signal that is necessary for preferential viral RNA packaging by coat protein disks is known and characterized since the 1980's. In this work, we have proposed a two-component system based on a Nicotiana bentamiana plants infection with packaging competent defective RNA (dRNA) and a helper virus RNA which provides all the components necessary for dRNA replication and packaging. The encapsidation signal in the helper virus sequence was removed to avoid formation of particles of incorrect length. Some of our helper viruses contained a coat protein with modified region of the particle's inner channel. This modification should allow specific binding of metal atoms within the core of the rod shaped particle. Several variants of dRNA and helper viruses were prepared to identify individual areas important for the replication, encapsidation and nanoparticle stability. We focused on the particle formation...

The action of 28 antibiotics and antimetabolites at varying concentrations was studied in coincidence with the bioassay of tobacco mosaic virus on detached bean leaves by using an ultrasonic method of inoculation developed by Lamborn, Cochran and Chidester. All tested chemicals were classified, in regard to the virus infection process, as inhibitory or stimulatory and their optimal concentrations in the inoculum were determined. Of the 28 antibiotics and antimetabolites studied, eight were found to stimulate while twenty inhibited TMV infection in bean leaves. From the results of these experiments, an attempt was made to characterize the nature of the cellular antiviral defense mechanism operating in the bean leaves. The results of this study may also be used in the development of a new TMV bioassay procedure with increased sensitivity.

Active disease resistance in plants is induced during the pathogen infection process that triggers multiple defense-related genes to establish broad-spectrum resistance. Several biotic and abiotic agents can mimic natural induced resistance (IR), categorized as systemic acquired (SAR) or induced systemic resistance (ISR). IR, triggered by acibenzolar-S-methyl (ASM) or plant growth-promoting rhizobacteria (PGPR), was evaluated on two-to-three types of tobacco in greenhouse and field studies. Tobacco mosaic virus (TMV) local lesion assays monitored induction and maintenance of ASM-induced SAR over a 21 day period via proportional reduction in the number of TMV local lesions between an untreated control and ASM-treated plants. Intraspecific variation in SAR was found among tobacco types; burley and flue-cured tobaccos responded by day 3, while oriental tobacco responded between day 3 and 6. The SAR signal was greatest between 6 and 15 days following ASM application, but IR was slightly evident even at 21 days after ASM application in all three tobacco types. Bottom and middle leaves responded similarly on all sample dates, but top leaves showed the weakest SAR response. Tobacco cyst nematode (TCN; Globodera tabacum solanacearum) is one of the most destructive pathogens of tobacco in Virginia. Among four PGPR combinations tested, a mixture of Bacillus amyloliquefaciens IN937a (GB99) and B. subtilis A13 (GB122) most consistently suppressed TCN reproduction in flue-cured and oriental tobacco. Application of ASM similarly reduced final numbers of TCN cysts, but also resulted in chlorosis, stunting, and lower plant fresh weight. GB99+GB122 also suppressed TCN development and reproduction in susceptible and resistant flue-cured cultivars, but reductions by ASM were less consistent. In a split-root trial, soil amendment with GB99+GB122 in one half of an oriental tobacco root system lowered final numbers of TCN more than did ASM. ASM exhibited undesirable effects in phytotoxicity trials in flue-cured and oriental tobacco, but GB99+GB122 was not phytotoxic. When oriental tobacco seedlings were grown in a GB99+GB122-treated soil-less media, a single application of 200 mg ASM/L one week after transplanting significantly suppressed TCN reproduction in the field without phytotoxicity. Further field research is needed to confirm this effect in flue-cured tobacco. / Ph. D.