Single-cell imaging of CAR T cell activity in vivo reveals extensive functional and anatomical heterogeneity

CAR T cells represent a potentially curative strategy for B cell malignancies. However, the outcome and dynamics of CAR T cell interactions in distinct anatomical sites are poorly understood. Using intravital imaging, we tracked interactions established by anti-CD19 CAR T cells in B cell lymphoma–bearing mice. Circulating targets trapped CAR T cells in the lungs, reducing their access to lymphoid organs. In the bone marrow, tumor apoptosis was largely due to CAR T cells that engaged, killed, and detached from their targets within 25 min. Notably, not all CAR T cell contacts elicited calcium signaling or killing while interacting with tumors, uncovering extensive functional heterogeneity. Mathematical modeling revealed that direct killing was sufficient for tumor regression. Finally, antigen-loss variants emerged in the bone marrow, but not in lymph nodes, where CAR T cell cytotoxic activity was reduced. Our results identify a previously unappreciated level of diversity in the outcomes ofCAR T cell interactions in vivo, with important clinical implications.Toxicolog

Un mécanisme de détection du quorum basé sur le métabolisme cellulaire participe à la résolution des réponses inflammatoires

Recruitment of immune cells during infection is essential to fueling the immune response but can also trigger immunopathology. A critical question is how the immune system can sense inflammation levels and self-adjust accordingly to limit tissue damage while removing the pathogen. During my Ph.D. I studied the self-resolving cutaneous infection with Leishmania major parasites where tissue damage arises when inflammation is allowed to become excessive. At the site of infection, the immune reaction is driven by recruited monocyte-derived cells that represents the major population of infected cells and are also actively involved in fighting the infection. They secrete pro-inflammatory cytokines but also produce nitric oxide (NO), critical to regulate the outcome of the infection: iNOS KO mice are susceptible and do not control the parasite load, subsequently developing severe tissue damage because of excessive immune cell infiltration. My work demonstrated that monocyte-derived cells at the site of infection are regulated by NO that limits their cellular respiration, lowers their energetic resources and consequently their activity in vivo. This regulation relies on tissue-wide NO diffusion and only exists when a sufficient cell density has been reached, revealing that monocyte-derived cells are endowed with a quorum sensing mechanism that adjusts their population size and activity in time and space to avoid immunopathology.Lors d’une infection, le recrutement de cellules immunitaires au site inflammatoire est nécessaire pour la lutte contre le pathogène mais peut également participer au déclenchement d’une immunopathologie. Il n’est pas encore clair aujourd’hui s’il existe des mécanismes permettant au système immunitaire de percevoir l’intensité de la réponse inflammatoire pour s’autoréguler afin d’éviter une importante destruction tissulaire tout en éliminant le pathogène. Pendant mes études doctorales, j’ai étudié l’infection asymptomatique par le parasite Leishmania major qui peut générer d’important dommages au tissu si l’inflammation devient excessive. La réaction immunitaire contre le parasite est contrôlée par des cellules dérivées de monocytes qui sont recrutées au site inflammatoire et y forment la principale population infectée en plus de participer activement à la réaction inflammatoire. Elles secrètent des cytokines pro-inflammatoires et produisent de l’oxyde nitrique (NO) qui est essentiel pour une issue favorable de la maladie. En effet, les souris déficientes pour l’enzyme iNOS (synthétisant le NO) contrôlent moins bien l’infection et développent systématiquement de graves symptômes associés à d’importants dommages tissulaires, causés par un recrutement incontrôlé de cellules immunitaires. Mon travail a montré que les cellules dérivées de monocytes sont régulées au site d’infection par le NO qui limite leur respiration cellulaire, diminuant ainsi leurs ressources énergétiques et leur activité in vivo. Ce mode de régulation nécessite la diffusion du NO à distance et n’existe que lorsque les cellules dérivées de monocytes sont densité suffisante. Ceci montre que ces cellules sont régulées par un mécanisme de détection du quorum, basé sur le métabolisme cellulaire, qui permet d’ajuster finement la quantité de cellules immunitaires actives dans l’espace et le temps pour éviter le développement d’une immunopathologie

Représentation et optimisation de maillage structuré par blocs à l'aide de systèmes multi-agents

This thesis deals with the representation and generation of block-structured hexahedral meshes. To date, there is no method for generating satisfactory block structures for any geometric domain. In practice, expert engineers generate these meshes using interactive software, which can take several weeks to complete. Moreover, adding modification operations in these interactive softwares is a delicate task to maintain the coherence of the block structure and its relationship with the geometric domain to be discretized. In order to improve this process, we first propose to define hexahedral mesh manipulation operations based on the use of the generalized map model. Then, by considering block structures obtained using the Polycube method, we provide methods for optimizing the topology of these structures to satisfy constraints of a geometric nature. We propose a first method in dimension 2, which considers a local approach to the problem based on the experience of engineers working with interactive software. We then propose a second method, this time using ant colony optimization meta-heuristics for leaf selection in dimension 3.Ce travail de thèse porte sur la représentation et la génération de maillages hexaédriques structurés par blocs. Il n'existe pas à ce jour de méthode permettant de générer des structures de blocs satisfaisantes pour n'importe quel domaine géométrique. En pratique, des ingénieurs experts génèrent ces maillages avec des logiciels interactifs, ce qui nécessite parfois plusieurs semaines de travail. De plus, l'ajout d'opérations de modification dans ces logiciels interactifs est un travail délicat pour maintenir la cohérence de la structure de blocs et sa relation avec le domaine géométrique à discrétiser. Afin d'améliorer ce processus, nous proposons tout d'abord de définir des opérations de manipulation de maillages hexaédriques se basant sur l'utilisation du modèle des cartes généralisées. Ensuite, en considérant des structures de blocs obtenues à l'aide de la méthode des Polycubes, nous fournissons des méthodes optimisant la topologie de ces structures pour satisfaire des contraintes de nature géométrique. Nous proposons ainsi une première méthode en dimension 2, qui considère une approche locale du problème en s'appuyant sur l'expérience des ingénieurs manipulant des logiciels interactifs. Puis nous proposons une seconde méthode utilisant cette fois la méta-heuristique d'optimisation par colonie de fourmis pour la sélection de feuillets en dimension 3

Quorum Sensing by Monocyte-Derived Populations

International audienceQuorum sensing is a type of cellular communication that was first described in bacteria, consisting of gene expression regulation in response to changes in cell-population density. Bacteria synthesize and secrete diffusive molecules called autoinducers, which concentration varies accordingly with cell density and can be detected by the producing cells themselves. Once autoinducer concentration reaches a critical threshold, all bacteria within the autoinducer-rich environment react by modifying their genetic expression and adopt a coordinated behavior (e.g., biofilm formation, virulence factor expression, or swarming motility). Recent advances highlight the possibility that such type of communication is not restricted to bacteria, but can exist among other cell types, including immune cells and more specifically monocyte-derived cells (1). For such cells, quorum sensing mechanisms may not only regulate their population size and synchronize their behavior at homeostasis but also alter their activity and function in unexpected ways during immune reactions. Although the nature of immune autoinducers and cellular mechanisms remains to be fully characterized, quorum sensing mechanisms in the immune system challenge our traditional conception of immune cell interactions and likely represent an important mode of communication at homeostasis or during an immune response. In this mini-review, we briefly present the prototypic features of quorum sensing in bacteria and discuss the existing evidence for quorum sensing within the immune system. Mainly, we review quorum sensing mechanisms among monocyte-derived cells, such as the regulation of inflammation by the density of monocyte-derived cells that produce nitric oxide and discuss the relevance of such models in the context of immune-related pathologies

Formal Definition of Hexahedral Blocking operations Using n-G-Maps

International audienceNowadays for real study cases, the generation of full block structured hexahedral meshes is mainly an interactive and very-time consuming process realized by highly-qualified engineers. To this purpose, they use interactive software where they handle and modify complex block structures with operations like block removal, block insertion, O-grid insertion, propagation of block splitting, propagation of meshing parameters along layers of blocks and so on. Such operations are error-prone and modifying or adding an operation is a very tedious work. In this work, we propose to formally define hexahedral block structures and main associated operations in the model of n-dimensional generalized map. This model provides topological invariant and a systematic handling of geometric data that allows us to ensure the expected robustness

Quorum sensing governs collective dendritic cell activation in vivo

We thank M. Guerin for help in image acquisition and the mouse facility and Technology Core of the Center for Translational Science (CRT) at Institut Pasteur for support in conducting the present studyInternational audienceDendritic cell (DC) activation by viral RNA sensors such as TLR3 and MDA-5 is critical for initiating antiviral immunity. Optimal DC activation is promoted by type I interferon (IFN) signaling which is believed to occur in either autocrine or paracrine fashion. Here, we show that neither autocrine nor paracrine type I IFN signaling can fully account for DC activation by poly(I:C) in vitro and in vivo. By controlling the density of type I IFN-producing cells in vivo, we establish that instead a quorum of type I IFN-producing cells is required for optimal DC activation and that this process proceeds at the level of an entire lymph node. This collective behavior, governed by type I IFN diffusion, is favored by the requirement for prolonged cytokine exposure to achieve DC activation. Furthermore, collective DC activation was found essential for the development of innate and adaptive immunity in lymph nodes. Our results establish how collective rather than cell-autonomous processes can govern the initiation of immune responses

A metabolism-based quorum sensing mechanism contributes to termination of inflammatory responses

International audienceRecruitment of immune cells with antimicrobial activities is essential to fight local infections but has the potential to trigger immunopathology. Whether the immune system has the ability to sense inflammation intensity and self-adjust accordingly to limit tissue damage remains to be fully established. During local infection with an intracellular pathogen, we have shown that nitric oxide (NO) produced by recruited monocyte-derived cells was essential to limit inflammation and cell recruitment. Mechanistically, we have provided evidence that NO dampened monocyte-derived cell cytokine and chemokine production by inhibiting cellular respiration and reducing cellular ATP:ADP ratio. Such metabolic control operated at the tissue level but only when a sufficient number of NO-producing cells reached the site of infection. Thus, NO production and activity act as a quorum sensing mechanism to help terminate the inflammatory response