QUANTITATIVE TRACKING OF CRYPTOSPORIDIUM INFECTION IN CELL CULTURE WITH CFSE

Immunofluorescence-based assays have been developed to detect and quantitate Cryptosporidium parvum infection in cell culture. Here, we describe a method that tracks and quantifies the early phase of attachment and invasion of C. parvum sporozoites using a fluorescent dye. Newly excysted sporozoites...

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Bibliographic Details
Published in:The Journal of parasitology 2006-12, Vol.92 (6), p.1350-1354
Main Authors: Feng, Hanping, Nie, Weijia, Bonilla, Ruben, Widmer, Giovanni, Sheoran, Abhineet, Tzipori, Saul
Format: Article
Language:English
Subjects:
Actins
Actins - metabolism
Animals
Biological and medical sciences
Cattle
Cell Line
Cell Line, Tumor
Cell membranes
Cryptosporidium - isolation & purification
Cryptosporidium - physiology
Cryptosporidium parvum
Cryptosporidium parvum - isolation & purification
Cryptosporidium parvum - physiology
Cytometry
Flow Cytometry
Fluoresceins
Fluorescent Dyes
Fundamental and applied biological sciences. Psychology
General aspects
General aspects and techniques. Study of several systematic groups. Models
Host-Parasite Interactions
Humans
Infections
Inoculation
Invertebrates
Microscopy, Confocal
Microscopy, Fluorescence
Oocysts
Parasite hosts
Parasites
Sporozoites
Staining and Labeling
Succinimides
T lymphocytes
THERAPEUTICS-DIAGNOSTICS
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Summary:Immunofluorescence-based assays have been developed to detect and quantitate Cryptosporidium parvum infection in cell culture. Here, we describe a method that tracks and quantifies the early phase of attachment and invasion of C. parvum sporozoites using a fluorescent dye. Newly excysted sporozoites were labeled with the amine-reactive fluorescein probe carboxyfluorescein diacetate succinimidyl esters (CFSE) using an optimized protocol. The initial invasion of cells by labeled parasites was detected with fluorescent or confocal microscopy. The infection of cells was quantified by flow cytometry. Comparative analysis of infection of cells with CFSE-labeled and unlabeled sporozoites showed that the infectivity of C. parvum was not affected by CFSE labeling. Quantitative analysis showed that C. parvum Iowa and MD isolates were considerably more invasive than Cryptosporidium hominis isolate TU502. Unlike immunofluorescent assays, CFSE labeling permitted the tracking of the initial invasion of C. parvum. Such an assay may be useful for studying the dynamics of host cell–parasite interaction and possibly for drug screening.
ISSN:0022-3395
1937-2345
DOI:10.1645/GE-853R.1