High‐Speed Cell Sorting of Infectious Trophic and Cystic Forms of <i>Pneumocystis carinii</i>

Martinez, Anna; Aliouat, El Moukhtar; Pottier, Muriel; Gantois, Nausicâa; Pinçon, Claire; Standaert-Vitse, Annie; Dei‐Cas, Eduardo; Aliouat-Denis, Cécile-Marie

doi:10.1111/j.1550-7408.2009.00423.x

Cited by 15 publications

(40 citation statements)

References 31 publications

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“…Athymic Pneumocystis -free Lou nu/nu rats (Institut Pasteur de Lille, Lille, France) were used as source of P. carinii organisms for all experiments [29]. Dexamethasone (Merck Sharp & Dohme Chibret, France) was administered to ten-week-old female or male nude rats for two weeks in the drinking water (1 mg/L).…”

Section: Methodsmentioning

confidence: 99%

Diamidines versus Monoamidines as Anti-Pneumocystis Agents: An in Vivo Study

et al. 2013

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Some compounds articulated around a piperazine or an ethylenediamine linker have been evaluated in vitro to determine their activity in the presence of a 3T6 fibroblast cell line and an axenic culture of Pneumocystis carinii, respectively. The most efficient antifungal derivatives, namely N,N′-bis(benzamidine-4-yl)ethane-1,2-diamine (compound 6, a diamidine) and N-(benzamidine-4-yl)-N′-phenylethane-1,2-diamine (compound 7, a monoamidine), exhibited no cytotoxicity and were evaluated in vivo in a rat model. Only the diamidine 6 emerged as a promising hit for further studies.

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Section: Methodsmentioning

confidence: 99%

Diamidines versus Monoamidines as Anti-Pneumocystis Agents: An in Vivo Study

et al. 2013

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“…The unavailability of continuous in vitro culture systems has hindered research aiming at clarifying the role of each life cycle stage in Pneumocystis proliferation and transmission (44). However, ultrastructure (25,99), short-term culture (2,19), and cell sorting approaches (56,57) led to quite heuristic life cycle hypotheses. Furthermore, using PcP animal models, molecular strategies and the exploring of Pneumocystis occurrence in wild mammals resulted in major advancements, like the notions of strong host species specificity (1,27,31,33,38,41) and coevolution (3,27,31,46).…”

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confidence: 99%

Characterizing Pneumocystis in the Lungs of Bats: Understanding Pneumocystis Evolution and the Spread of Pneumocystis Organisms in Mammal Populations

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Pinçon

Aliouat-Denis

et al. 2012

Appl Environ Microbiol

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Bats belong to a wide variety of species and occupy diversified habitats, from cities to the countryside. Their different diets (i.e., nectarivore, frugivore, insectivore, hematophage) lead Chiroptera to colonize a range of ecological niches. These flying mammals exert an undisputable impact on both ecosystems and circulation of pathogens that they harbor. Pneumocystis species are recognized as major opportunistic fungal pathogens which cause life-threatening pneumonia in severely immunocompromised or weakened mammals. Pneumocystis consists of a heterogeneous group of highly adapted host-specific fungal parasites that colonize a wide range of mammalian hosts. In the present study, 216 lungs of 19 bat species, sampled from diverse biotopes in the New and Old Worlds, were examined. Each bat species may be harboring a specific Pneumocystis species. We report 32.9% of Pneumocystis carriage in wild bats (41.9% in Microchiroptera). Ecological and behavioral factors (elevation, crowding, migration) seemed to influence the Pneumocystis carriage. This study suggests that Pneumocystis-host association may yield much information on Pneumocystis transmission, phylogeny, and biology in mammals. Moreover, the link between genetic variability of Pneumocystis isolated from populations of the same bat species and their geographic area could be exploited in terms of phylogeography.

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“…High-speed cell sorting was our chosen strategy to purify P. carinii organisms from host cell debris and to achieve stage-to-stage separation with high purity (Martinez et al 2009). A polyclonal antibody produced in our laboratory was used to label the whole P. carinii population.…”

Section: How To Investigate the Pneumocystis Cell Cycle?mentioning

confidence: 99%

“…No other ultrastructural alterations were noted. Separation of cyst forms from trophic forms was achieved by flow cytometry with a purity of 99.6 ± 0.3% in 27 independent cell sorting experiments (Figs 2-6) (Martinez et al 2009). …”

Section: How To Investigate the Pneumocystis Cell Cycle?mentioning

confidence: 99%