A Quantitative 3D Motility Analysis of Trypanosoma brucei by Use of Digital In-line Holographic Microscopy

Weiße, Sebastian; Heddergott, Niko; Heydt, Matthias; Pflästerer, Daniel; Maier, Timo; Haraszti, Tamás; Grunze, M.; Engstler, Markus; Rosenhahn, Axel

doi:10.1371/journal.pone.0037296

Cited by 31 publications

(32 citation statements)

References 32 publications

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“…brucei typically contain cells with a variety of different swimming behaviours varying from persistent directional swimming to tumbling [56, 57]. Cell microenvironment affects this motility, whereby trypanosomes cultured in the low viscosities of tissue culture media predominantly tumble [54].…”

Section: Discussionmentioning

confidence: 99%

Blocking Synthesis of the Variant Surface Glycoprotein Coat in Trypanosoma brucei Leads to an Increase in Macrophage Phagocytosis Due to Reduced Clearance of Surface Coat Antibodies

et al. 2016

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The extracellular bloodstream form parasite Trypanosoma brucei is supremely adapted to escape the host innate and adaptive immune system. Evasion is mediated through an antigenically variable Variant Surface Glycoprotein (VSG) coat, which is recycled at extraordinarily high rates. Blocking VSG synthesis triggers a precytokinesis arrest where stalled cells persist for days in vitro with superficially intact VSG coats, but are rapidly cleared within hours in mice. We therefore investigated the role of VSG synthesis in trypanosome phagocytosis by activated mouse macrophages. T. brucei normally effectively evades macrophages, and induction of VSG RNAi resulted in little change in phagocytosis of the arrested cells. Halting VSG synthesis resulted in stalled cells which swam directionally rather than tumbling, with a significant increase in swim velocity. This is possibly a consequence of increased rigidity of the cells due to a restricted surface coat in the absence of VSG synthesis. However if VSG RNAi was induced in the presence of anti-VSG221 antibodies, phagocytosis increased significantly. Blocking VSG synthesis resulted in reduced clearance of anti-VSG antibodies from the trypanosome surface, possibly as a consequence of the changed motility. This was particularly marked in cells in the G2/ M cell cycle stage, where the half-life of anti-VSG antibody increased from 39.3 ± 4.2 seconds to 99.2 ± 15.9 seconds after induction of VSG RNAi. The rates of internalisation of bulk surface VSG, or endocytic markers like transferrin, tomato lectin or dextran were not significantly affected by the VSG synthesis block. Efficient elimination of anti-VSG-antibody complexes from the trypanosome cell surface is therefore essential for trypanosome evasion of macrophages. These experiments highlight the essentiality of high rates of VSG recycling for the rapid removal of host opsonins from the parasite surface, and identify this process as a key parasite virulence factor during a chronic infection.

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Section: Discussionmentioning

confidence: 99%

Blocking Synthesis of the Variant Surface Glycoprotein Coat in Trypanosoma brucei Leads to an Increase in Macrophage Phagocytosis Due to Reduced Clearance of Surface Coat Antibodies

et al. 2016

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“…Digital in-line holographic microscopy (DIHM), as schematically shown in Fig. 3, is a technique capable to capture fast, three-dimensional motions by recording a movie of coherent far-field diffraction patterns of the sample volume [55][56][57][58][59]. A coherent laser beam is focused on a small pinhole to create a divergent light cone, which illuminates the volume of interest.…”

Section: Biofouling Research: the Quest For Environmentallymentioning

confidence: 99%

Surface Sensing and Settlement Strategies of Marine Biofouling Organisms

Rosenhahn

Sendra

2012

Biointerphases

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This review article summarizes some recent insights into the strategies used by marine organisms to select surfaces for colonization. While larger organisms rely on their sensory machinery to select surfaces, smaller microorganisms developed less complex but still effective ways to probe interfaces. Two examples, zoospores of algae and barnacle larvae, are discussed and both appear to have build-in test mechanisms to distinguish surfaces with different physicochemical properties. Some systematic studies on the influence of surface cues on exploration, settlement and adhesion are summarized. The intriguing notion that surface colonization resembles a parallelized surface sensing event is discussed towards its complementarity with conventional surface analytical tools. The strategy to populate only selected surfaces seems advantageous as waves, currents and storms constantly challenge adherent soft and hard fouling organism.

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“…Early work by Walker () demonstrated that motility is driven by a tractile flagellar beat that initiates at the tip of the flagellum and travels to the flagellum base, the opposite of what is seen in most other eukaryotic flagella. More recent analyses of T. brucei motility using high‐speed, high‐resolution video microscopy provided adjustments to the original ‘auger‐like’ description and further emphasized that parasite motility is a prominent feature of these pathogens in the blood (Rodriguez et al ., ; Uppaluri et al ., ; Heddergott et al ., ; Weisse et al ., ).…”

Section: Introductionmentioning

confidence: 97%

Mouse infection and pathogenesis byTrypanosoma bruceimotility mutants

et al. 2014

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The flagellum of Trypanosoma brucei is an essential and multifunctional organelle that drives parasite motility and is receiving increased attention as a potential drug target. In the mammalian host, parasite motility is suspected to contribute to infection and disease pathogenesis. However, it has not been possible to test this hypothesis owing to lack of motility mutants that are viable in the bloodstream life cycle stage that infects the mammalian host. We recently identified a bloodstream-form motility mutant in 427-derived T. brucei in which point mutations in the LC1 dynein subunit disrupt propulsive motility but do not affect viability. These mutants have an actively beating flagellum, but cannot translocate. Here we demonstrate that the LC1 point mutant fails to show enhanced cell motility upon increasing viscosity of the surrounding medium, which is a hallmark of wild type T. brucei, thus indicating that motility of the mutant is fundamentally altered compared to wild type cells. We next used the LC1 point mutant to assess the influence of trypanosome motility on infection in mice. We surprisingly found that disrupting parasite motility has no discernible effect on T. brucei bloodstream infection. Infection time-course, maximum parasitemia, number of waves of parasitemia, clinical features and disease outcome are indistinguishable between motility mutant and control parasites. Our studies provide an important step toward understanding the contribution of parasite motility to infection and a foundation for future investigations of T. brucei interaction with the mammalian host.

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A Quantitative 3D Motility Analysis of Trypanosoma brucei by Use of Digital In-line Holographic Microscopy

Cited by 31 publications

References 32 publications

Blocking Synthesis of the Variant Surface Glycoprotein Coat in Trypanosoma brucei Leads to an Increase in Macrophage Phagocytosis Due to Reduced Clearance of Surface Coat Antibodies

Blocking Synthesis of the Variant Surface Glycoprotein Coat in Trypanosoma brucei Leads to an Increase in Macrophage Phagocytosis Due to Reduced Clearance of Surface Coat Antibodies

Surface Sensing and Settlement Strategies of Marine Biofouling Organisms

Mouse infection and pathogenesis byTrypanosoma bruceimotility mutants

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