Flow Cytometric Quantification of Toxoplasma gondii Cellular Infection and Replication

Gay-Andrieu, Françoise; Cozon, G.; Ferrandiz, Josette; Kahi, Sandrine; Peyron, François

doi:10.2307/3285793

Cited by 19 publications

(17 citation statements)

References 21 publications

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“…Toxoplasma gondii, causing severe encephalitis in immunocompromised hosts, is able to attach to and invade a wide range of vertebrate nucleated cells (Sibley, 1995). The ability of T. gondii to invade microglia (Fischer et al, 1997) and BMEC (Gay-Andrieu et al, 1999) suggest that T. gondii may enter the CNS by transmigrating the BBB. In sharp contrast to T. gondii, malaria Plasmodium parasites primarily invade reticulocytes, particularly erythrocytes.…”

Section: Strategies Of Cns Invasion By Fungi and Parasitesmentioning

confidence: 99%

Molecular and cellular mechanisms for microbial entry into the CNS

Zhang¹,

Tuomanen²

1999

J Neurovirol

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A number of pathogenic microbes including neuroinvasive viruses, bacteria and parasites are capable of entry into the central nervous system (CNS) and cause a variety of clinical manifestations. The cellular and molecular mechanisms for the CNS invasion have been extensively studied in the last two decades. Viruses invade neurons and thereby cause encephalitis or peripheral neuritis, while bacteria enter the cerebrospinal¯uid (CSF) and cause meningitis. In contrast, the mechanisms for parasitic neuroinvasion are much more complex and less clear. The capabilities that enable these elite subsets of pathogens to engineer uptake into the CNS will be the subject of this review.

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Section: Strategies Of Cns Invasion By Fungi and Parasitesmentioning

confidence: 99%

Molecular and cellular mechanisms for microbial entry into the CNS

Zhang¹,

Tuomanen²

1999

J Neurovirol

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“…However, the use of FCM in parasitology has been largely restricted to the study of extracellular parasites. Exceptions are the Plasmodium spp., which can be specifically labeled with fluorescent DNA-binding dyes (18), and cytometric assays for the quantitation of intracellular Leishmania and Toxoplasma infections (12,15,23). In this report, we describe the use of FCM as a tool to allow the determination of intracellular microsporidia in various cell lines and humanderived macrophages.…”

Section: Discussionmentioning

confidence: 99%

“…FCM has found a broad range of applications in bacteriology and parasitology and has been used in studies of various parasitic protozoa such as Plasmodium, Toxoplasma, Eimeria, Leishmania, free living amoeba, Cryptosporidium, Giardia, and microsporidia (12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22). It has been used to quantitate and determine the viability of E. cuniculi spores from culture (5), to isolate E. bieneusi spores from stool samples (19), to compare the nucleic acid contents of microsporidian spores isolated from fish (20), and to discriminate between the spores of the three species of Encephalitozoon (22).…”

Section: Discussionmentioning

confidence: 99%

Quantitation of microsporidia in cultured cells by flow cytometry

et al. 2004

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Background: Microsporidia are obligate intracellular protozoan parasites that emerged as major opportunistic pathogens in humans since the onset of the AIDS pandemic. In the present study, we investigated whether FCM is a useful method for the quantitation of intracellular microsporidian spores in cultured cells. Methods: Microsporidia (Encephalitozoon cuniculi) were grown in cell cultures and various cell-lines were coincubated with microsporidian spores at different multiplicities of infection, as well as for different periods of time. After permeabilization of the cells, intracellular spores were stained with a polyclonal anti-E. cuniculi serum and a FITC-labeled secondary antibody. Stained cells were analyzed on a flow cytometer and results were compared with those of fluorescence microscopy.

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“…These techniques have largely been used in the fields of medical science and cell biology (10). Recently, flow cytometry has been introduced in the medical field of parasitology for the study of human cells infected with protozoan parasites (11,18,19). Many reliable techniques are now extensively used to detect parasites of the genus Plasmodium in human red blood cells (22,24), and various fluorescent probes or monoclonal antibodies have been developed for use in the exploration of interactions between parasites and host cells (21,24).…”

Section: Discussionmentioning

confidence: 99%

Flow Cytometric Detection of Leishmania Parasites in Human Monocyte-Derived Macrophages: Application to Antileishmanial-Drug Testing

Giorgio¹,

Ridoux²,

Delmas³

et al. 2000

Antimicrob Agents Chemother

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A flow cytometric technique was developed for detection of amastigotes of the protozoan Leishmania infantum in human nonadherent monocyte-derived macrophages. The cells were fixed and permeabilized with paraformaldehyde-ethanol, and intracellular amastigotes were labeled with Leishmania lipophosphoglycan-specific monoclonal antibody. Results showed that flow cytometry provided accurate quantification of the infection rates in human macrophages compared to the rates obtained by the conventional microscopic technique, with the advantage that a large number of cells could be analyzed rapidly. The results demonstrated, moreover, that labeling of intracellular amastigotes could reliably be used to evaluate the antileishmanial activities of conventional drugs such as meglumine antimoniate, amphotericin B, pentamidine, and allopurinol. They also established that various Leishmania species (L. mexicana, L. donovani) could be detected by this technique in other host-cell models such as mouse peritoneal macrophages and suggested that the flow cytometric method could be a valid alternative to the conventional method.Leishmaniases are vector-borne parasitic diseases caused by protozoa of the genus Leishmania. These obligate intracellular parasites replicate in the parasitophorus vacuoles of macrophages (3, 6) and produce life-threatening disorders widely distributed in tropical, subtropical, and Mediterranean countries (Division of Control of Tropical Diseases, World Health Organization, http://www.who.int/ctd/html/leish.html). Several clinical syndromes are subsumed under the term leishmaniasis according to the localization of the parasites in mammalian tissues, notably, visceral, cutaneous, and mucosal leishmaniases. Parasites of the species Leishmania infantum, identified as a causative agent of the visceral form of leishmaniasis, have been shown to be endemic in France (Division of Control of Tropical Diseases, World Health Organization).Since the 1940s, the generally accepted therapy for all forms of leishmaniasis consisted of pentavalent antimonial agents, used as first-line clinical treatment (7, 23); however, antileishmanial therapy has been a bewildering subject largely because of the complexity of the disease. Moreover, the recent appearance of clinical resistance to meglumine antimoniate (8,13) and the small number of efficient second-line antileishmanial drugs raised the necessity to develop new methods for the rapid assessment of the antileishmanial activities of chemical compounds.Various in vitro host-cell models such as mouse peritoneal macrophages (4), human monocytes (U-937) (16), or Chinese hamster ovary (CHO) cells (26) have been created to study infection of mammalian cells with Leishmania parasites and to test the antileishmanial activities of new molecules. In these models, infection rates were usually measured by microscopic examination of adherent cells. In the present study, we investigated the possibility of using flow cytometry for the rapid and reliable detection of possible antileishmanial drug...

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Flow Cytometric Quantification of Toxoplasma gondii Cellular Infection and Replication

Cited by 19 publications

References 21 publications

Molecular and cellular mechanisms for microbial entry into the CNS

Molecular and cellular mechanisms for microbial entry into the CNS

Quantitation of microsporidia in cultured cells by flow cytometry

Flow Cytometric Detection of Leishmania Parasites in Human Monocyte-Derived Macrophages: Application to Antileishmanial-Drug Testing

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