Enzyme immunoassay detection of Cryptosporidium parvum inhibition by sinefungin in sporozoite infected HCT-8 enterocytic cells.
Complete parasite development was obtained in differentiated human enterocytic HCT-8 cells infected at confluence with Cryptosporidium parvum sporozoites, and evaluated in a quantitative enzyme immunoassay. Forty-eight hours after infection, a linear correlation was found between optical density values and the number of parasites determined in an immunofluorescent assay. Sinefungin exerted an inhibitory effect when added within 4 h after sporozoite addition to HCT-8 cultures (MIC50 = 38 mumol L-1), while the inhibitory effects of paromomycin and pentamidine dimethanesulfonate were also easily detected (MIC50 = 0.87 mumol L-1 and 0.27 mumol L-1, respectively). Except for high pentamidine dimethanesulfonate concentrations, no alteration in optical microscopy morphology or trypan blue exclusion of HCT-8 cells was observed in the presence of anticryptosporidial agents, which suggests that they were primarily active against developing parasites. Data suggest that EIA detection of C. parvum development in sporozoite-infected HCT-8 cells provides an accurate and convenient model for in vitro evaluation of parasite infectivity, growth and response to anticryptosporidial agents.
Laboratoire d'Immunologie et Immunopathologie, UPRES-EA 2128, CHU, Caen, France.
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G
Gargala
A
A
Delaunay
L
L
Favennec
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P
Brasseur
J J
JJ
Ballet
D000241:Adenosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000563:Amebicides / Q000494:pharmacology; D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D002460:Cell Line; D016785:Cryptosporidium parvum / Q000187:drug effects* / Q000302:isolation & purification; D004305:Dose-Response Relationship, Drug; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D010270:Parasite Egg Count; D010303:Paromomycin / Q000494:pharmacology; D010419:Pentamidine / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors
D000563:Amebicides; D000981:Antiprotozoal Agents; D010303:Paromomycin; D010419:Pentamidine; D000241:Adenosine; C006235:sinefungin