QUANTITATIVE TRACKING OF CRYPTOSPORIDIUM INFECTION IN CELL CULTURE WITH CFSE

• 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
• ISSN: 0022-3395; 1937-2345
• DOI: 10.1645/GE-853R.1

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.