Document Detail


Quantitative tracking of Cryptosporidium infection in cell culture with CFSE.
MedLine Citation:
PMID:  17304819     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Hanping Feng; Weijia Nie; Ruben Bonilla; Giovanni Widmer; Abhineet Sheoran; Saul Tzipori
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  The Journal of parasitology     Volume:  92     ISSN:  0022-3395     ISO Abbreviation:  J. Parasitol.     Publication Date:  2006 Dec 
Date Detail:
Created Date:  2007-02-19     Completed Date:  2007-03-01     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  7803124     Medline TA:  J Parasitol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1350-4     Citation Subset:  IM    
Affiliation:
Division of Infectious Diseases, Tufts University, Cummings School of Veterinary Medicine, North Grafton, Massachusetts 01536, USA.
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MeSH Terms
Descriptor/Qualifier:
Actins / metabolism
Animals
Cattle
Cell Line
Cell Line, Tumor
Cryptosporidium / isolation & purification*,  physiology
Cryptosporidium parvum / isolation & purification,  physiology
Flow Cytometry
Fluoresceins*
Fluorescent Dyes*
Host-Parasite Interactions
Humans
Microscopy, Confocal
Microscopy, Fluorescence
Staining and Labeling
Succinimides*
Grant Support
ID/Acronym/Agency:
N01 AI25466/AI/NIAID NIH HHS; R01 AI05471/AI/NIAID NIH HHS; RR017383/RR/NCRR NIH HHS
Chemical
Reg. No./Substance:
0/5-(6)-carboxyfluorescein diacetate succinimidyl ester; 0/Actins; 0/Fluoresceins; 0/Fluorescent Dyes; 0/Succinimides

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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