An Historical Perspective on How Advances in Microscopic Imaging Contributed to Understanding the<i>Leishmania</i>Spp. and<i>Trypanosoma cruzi</i>Host-Parasite Relationship

Florentino, Pilar Tavares Veras; Real, Fernando; Bonfim-Melo, Alexis; Orikaza, Cristina Mary; Ferreira, Éden Ramalho; Pessoa, Carina Carraro; Lima, Beatryz Rafaela Santos; Sasso, Gisela Rodrigues da Silva; Mortara, Renato Arruda

doi:10.1155/2014/565291

Cited by 9 publications

(8 citation statements)

References 101 publications

(126 reference statements)

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“…In the case of parasites, green fluorescent protein (GFP) originally purified from the jellyfish ( Aequoria victoria ) has been successfully expressed in several protozoan intracellular parasites such as Leishmania species [10–12], Trypanosoma [13–14], Enatomoeba [15], Plasmodium [16] and Toxoplasma [17], and more recently as a dual combination with other fluorescent reporters (for example with mCherry) to follow the developmental stages of T. gondii [18]. FPs have been also successfully used for deciphering pathogenesis of bacterial intracellular infection (some examples are given in [19–24]).…”

Section: Use Of External and Internal Fluorescent Reporters For Inmentioning

confidence: 99%

Imaging flow cytometry analysis of intracellular pathogens

Haridas

Ranjbar

Vorobjev

et al. 2017

Methods

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Imaging flow cytometry has been applied to address questions in infection biology, in particular, infections induced by intracellular pathogens. This methodology, which utilizes specialized analytic software makes it possible to analyze hundreds of quantified features for hundreds of thousands of individual cellular or subcellular events in a single experiment. Imaging flow cytometry analysis of host cell-pathogen interaction can thus quantitatively addresses a variety of biological questions related to intracellular infection, including cell counting, internalization score, and subcellular patterns of co-localization. Here, we provide an overview of recent achievements in the use of fluorescently labeled prokaryotic or eukaryotic pathogens in human cellular infections in analysis of host-pathogen interactions. Specifically, we give examples of Imagestream-based analysis of cell lines infected with Toxoplasma gondii or Mycobacterium tuberculosis. Furthermore, we illustrate the capabilities of imaging flow cytometry using a combination of standard IDEAS™ software and the more recently developed Feature Finder algorithm, which is capable of identifying statistically significant differences between researcher-defined image galleries. We argue that the combination of imaging flow cytometry with these software platforms provides a powerful new approach to understanding host control of intracellular pathogens.

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Section: Use Of External and Internal Fluorescent Reporters For Inmentioning

confidence: 99%

Imaging flow cytometry analysis of intracellular pathogens

Haridas

Ranjbar

Vorobjev

et al. 2017

Methods

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“…Since the discovery of Chagas' disease in 1909 (Chagas, ), major leaps in Trypanosoma cruzi biology have been achieved and have resulted in gradually improving advanced techniques for direct parasite observation (Florentino et al , ). Optical and electron microscopy (De Souza, ) are the most commonly used advanced techniques.…”

Section: Introductionmentioning

confidence: 99%

Trypanosoma cruzi : single cell live imaging inside infected tissues

Ferreira

Orikaza

Cordero

et al. 2016

Cellular Microbiology

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SummaryAlthough imaging the live Trypanosoma cruzi parasite is a routine technique in most laboratories, identification of the parasite in infected tissues and organs has been hindered by their intrinsic opaque nature. We describe a simple method for in vivo observation of live single‐cell Trypanosoma cruzi parasites inside mammalian host tissues. BALB/c or C57BL/6 mice infected with DsRed‐CL or GFP‐G trypomastigotes had their organs removed and sectioned with surgical blades. Ex vivo organ sections were observed under confocal microscopy. For the first time, this procedure enabled imaging of individual amastigotes, intermediate forms and motile trypomastigotes within infected tissues of mammalian hosts.

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“…However, the steps of the intracellular life cycle in mammalian hosts that involve the obligatory egress of Leishmania amastigote forms from host cells in order to the spread to new host cells and other tissues (tropism) and organisms are likely the least known aspect of the biology of this parasitic protozoan. A search of the early literature revealed that authors emphasized a 'lytic' cycle for this parasite, mainly based on histopathological observations fragmented in space and time (Theodorides, 1997;Dedet, 2007;Florentino et al, 2014). The preferential, almost exclusive, presence of oval-shaped parasites inside macrophages was the most puzzling aspect of leishmaniasis for the first histopathologists who studied the organism (Wright, 1903;Christophers, 1904), highlighting the uniqueness of Leishmania cell infection and supporting a concept of a specialized parasite, with a limited repertoire of cells able to host them.…”

Section: Introductionmentioning

confidence: 99%

Cell‐to‐cell transfer ofLeishmania amazonensisamastigotes is mediated by immunomodulatoryLAMP‐rich parasitophorous extrusions

et al. 2014

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The last step of Leishmania intracellular life cycle is the egress of amastigotes from the host cell and their uptake by adjacent cells. Using multidimensional live imaging of long-term-infected macrophage cultures we observed that Leishmania amazonensis amastigotes were transferred from cell to cell when the donor host macrophage delivers warning signs of imminent apoptosis. They were extruded from the macrophage within zeiotic structures (membrane blebs, an apoptotic feature) rich in phagolysosomal membrane components. The extrusions containing amastigotes were selectively internalized by vicinal macrophages and the rescued amastigotes remain viable in recipient macrophages. Host cell apoptosis induced by micro-irradiation of infected macrophage nuclei promoted amastigotes extrusion, which were rescued by non-irradiated vicinal macrophages. Using amastigotes isolated from LAMP1/LAMP2 knockout fibroblasts, we observed that the presence of these lysosomal components on amastigotes increases interleukin 10 production. Enclosed within host cell membranes, amastigotes can be transferred from cell to cell without full exposure to the extracellular milieu, what represents an important strategy developed by the parasite to evade host immune system.

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An Historical Perspective on How Advances in Microscopic Imaging Contributed to Understanding theLeishmaniaSpp. andTrypanosoma cruziHost-Parasite Relationship

Cited by 9 publications

References 101 publications

Imaging flow cytometry analysis of intracellular pathogens

Imaging flow cytometry analysis of intracellular pathogens

Trypanosoma cruzi : single cell live imaging inside infected tissues

Cell‐to‐cell transfer ofLeishmania amazonensisamastigotes is mediated by immunomodulatoryLAMP‐rich parasitophorous extrusions

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