Macrophage-pathogen interactions in infectious diseases: new therapeutic insights from the zebrafish host model

Studying macrophage biology in the context of a whole living organism provides unique possibilities to understand the contribution of this extremely dynamic cell subset in the reaction to infections, and has revealed the relevance of cellular and molecular processes that are fundamental to the cell-mediated innate immune response. In particular, various recently established zebrafish infectious disease models are contributing substantially to our understanding of the mechanisms by which different pathogens interact with macrophages and evade host innate immunity. Transgenic zebrafish lines with fluorescently labeled macrophages and other leukocyte populations enable non-invasive imaging at the optically transparent early life stages. Furthermore, there is a continuously expanding availability of vital reporters for subcellular compartments and for probing activation of immune defense mechanisms. These are powerful tools to visualize the activity of phagocytic cells in real time and shed light on the intriguing paradoxical roles of these cells in both limiting infection and supporting the dissemination of intracellular pathogens. This Review will discuss how several bacterial and fungal infection models in zebrafish embryos have led to new insights into the dynamic molecular and cellular mechanisms at play when pathogens encounter host macrophages. We also describe how these insights are inspiring novel therapeutic strategies for infectious disease treatment

Aangeboren immuniteit in beeld

Oratie uitgesproken door Prof.dr. A.H. Meijer bij de aanvaarding van het ambt van op het gebied van de Immunobiologie aan de Universiteit Leiden op maandag 21 september 2015Animal science

Agrobacterium-mediated transformation of the terpenoid indole alkaloid-producing plant species Tabernaemontana pandacaqui

Plants of the Apocynaceae family produce a wide range of terpenoid indole alkaloids (TIAs) which have important pharmaceutical applications. Studies of the molecular mechanisms controlling TIA biosynthesis may eventually provide possibilities to improve product yield by genetic modification of plants or cell cultures. However, these studies suffer from the lack of transformation/regeneration protocols for Apocynaceae plants. We chose to study the feasibility of Agrobacterium tumefaciens-mediated transformation of Tabernaemontana pandacaqui, because of the availability of an efficient regeneration procedure for this member of the Apocynaceae family. A procedure to produce transgenic T. pandacaqui plants was established, albeit with low efficiency. Transgenic expression was demonstrated of an intron-containing β-glucuronidase reporter gene and of a gene coding for the TIA biosynthetic enzyme strictosidine synthase from Catharanthus roseus, another Apocynaceae species.info:eu-repo/semantics/publishedVersio

Agrobacterium-mediated transformation of the terpenoid indole alkaloid-producing plant species Tabernaemontana pandacaqui

Plants of the Apocynaceae family produce a wide range of terpenoid indole alkaloids (TIAs) which have important pharmaceutical applications. Studies of the molecular mechanisms controlling TIA biosynthesis may eventually provide possibilities to improve product yield by genetic modification of plants or cell cultures. However, these studies suffer from the lack of transformation/regeneration protocols for Apocynaceae plants. We chose to study the feasibility of Agrobacterium tumefaciens-mediated transformation of Tabernaemontana pandacaqui, because of the availability of an efficient regeneration procedure for this member of the Apocynaceae family. A procedure to produce transgenic T. pandacaqui plants was established, albeit with low efficiency. Transgenic expression was demonstrated of an intron-containing β-glucuronidase reporter gene and of a gene coding for the TIA biosynthetic enzyme strictosidine synthase from Catharanthus roseus, another Apocynaceae species.info:eu-repo/semantics/publishedVersio

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

Animal science

Aspergillus fumigatus establishes infection in zebrafish by germination of phagocytized conidia, while Aspergillus niger relies on extracellular germination

Among opportunistically pathogenic filamentous fungi of the Aspergillus genus, Aspergillus fumigatus stands out as a drastically more prevalent cause of infection than others. Utilizing the zebrafish embryo model, we applied a combination of non-invasive real-time imaging and genetic approaches to compare the infectious development of A. fumigatus with that of the less pathogenic A. niger. We found that both species evoke similar immune cell migratory responses, but A. fumigatus is more efficiently phagocytized than A. niger. Though efficiently phagocytized, A. fumigatus conidia retains the ability to germinate and form hyphae from inside macrophages leading to serious infection even at relatively low infectious burdens. By contrast, A. niger appears to rely on extracellular germination, and rapid hyphal growth to establish infection. Despite these differences in the mechanism of infection between the species, galactofuranose mutant strains of both A. fumigatus and A. niger display attenuated pathogenesis. However, deficiency in this cell wall component has a stronger impact on A. niger, which is dependent on rapid extracellular hyphal growth. In conclusion, we uncover differences in the interaction of the two fungal species with innate immune cells, noticeable from very early stages of infection, which drive a divergence in their route to establishing infections

Aspergillus fumigatus establishes infection in zebrafish by germination of phagocytized conidia, while Aspergillus niger relies on extracellular germination

Among opportunistically pathogenic filamentous fungi of the Aspergillus genus, Aspergillus fumigatus stands out as a drastically more prevalent cause of infection than others. Utilizing the zebrafish embryo model, we applied a combination of non-invasive real-time imaging and genetic approaches to compare the infectious development of A. fumigatus with that of the less pathogenic A. niger. We found that both species evoke similar immune cell migratory responses, but A. fumigatus is more efficiently phagocytized than A. niger. Though efficiently phagocytized, A. fumigatus conidia retains the ability to germinate and form hyphae from inside macrophages leading to serious infection even at relatively low infectious burdens. By contrast, A. niger appears to rely on extracellular germination, and rapid hyphal growth to establish infection. Despite these differences in the mechanism of infection between the species, galactofuranose mutant strains of both A. fumigatus and A. niger display attenuated pathogenesis. However, deficiency in this cell wall component has a stronger impact on A. niger, which is dependent on rapid extracellular hyphal growth. In conclusion, we uncover differences in the interaction of the two fungal species with innate immune cells, noticeable from very early stages of infection, which drive a divergence in their route to establishing infections

Temperature Directs the Majority-Rules Principle in Supramolecular Copolymers Driven by Triazine–Benzene Interactions

Supramolecular copolymers have typically been studied in the extreme cases, such as self-sorting or highly mixed copolymer systems, while the intermediate systems have been less understood. We have reported the temperature-dependent microstructure in copolymers of triazine- and benzene-derivatives based on charge-transfer interactions with a highly alternating microstructure at low temperatures. Here, we investigate the temperature-dependent copolymerization further and increase the complexity by combining triazine- and benzene-derivatives with opposite preferred helicities. In this case, intercalation of the benzene-derivative into the triazine-derivative assemblies causes a helical inversion. The inversion of the net helicity was rationalized by comparing the mismatch penalties of the individual monomers, which indicated that the benzene-derivative dictates the helical screw-sense of the supramolecular copolymers. Surprisingly, this was not reflected in further investigations of slightly modified triazine- and benzene-derivatives, thus highlighting that the outcome is a subtle balance between structural features, where small differences can be amplified due to the competitive nature of the interactions. Overall, these findings suggest that the temperature-dependent microstructure of triazine- and benzene-based supramolecular copolymers determines the copolymer helicity of the presented system in a similar way as the mixed majority-rules phenomenon.</p

A fresh look at mycobacterial pathogenicity with the zebrafish host model

The zebrafish has earned its place among animal models to study tuberculosis and other infections caused by pathogenic mycobacteria. This model host is especially useful to study the role of granulomas, the inflammatory lesions characteristic of mycobacterial disease. The optically transparent zebrafish larvae provide a window on the initial stages of granuloma development in the context of innate immunity. Application of fluorescent dyes and transgenic markers enabled real-time visualization of how innate immune mechanisms, such as autophagy and inflammasomes, are activated in infected macrophages and how propagating calcium signals drive communication between macrophages during granuloma formation. A combination of imaging, genetic, and chemical approaches has revealed that the interplay between macrophages and mycobacteria is the main driver of tissue dissemination and granuloma development, while neutrophils have a protective function in early granulomas. Different chemokine signaling axes, conserved between humans and zebrafish, have been shown to recruit macrophages permissive to mycobacterial growth, control their microbicidal capacity, drive their spreading and aggregation, and mediate granuloma vascularization. Finally, zebrafish larvae are now exploited to explore cell death processes, emerging as crucial factors in granuloma expansion. In this review, we discuss recent advances in the understanding of mycobacterial pathogenesis contributed by zebrafish models.Animal science

A quantitative in vivo assay for craniofacial developmental toxicity of histone deacetylases

Animal science