Infection of human and bovine epithelial cells with Cryptosporidium andersoni induces apoptosis and disrupts tight junctional ZO-1: effects of epidermal growth factor

Abstract

The effects of Cryptosporidium andersoni on human or bovine epithelia are poorly defined. Epidermal growth factor inhibits colonisation of the gastrointestinal epithelium with bacteria and the enteric protozoan parasite Giardia lamblia. This study characterised whether C. andersoni infects human or bovine epithelial cells in vitro, assessed its impact on apoptosis and tight junctional Zonula–Occludens-1, and determined whether these effects may be altered by epidermal growth factor. Monolayers of human colonic CaCo 2 cells, SCBN (non-malignant small intestinal epithelial cells), and Madin Darby bovine kidney epithelial cell lines (MDBK and NBL-1) were grown to confluency in Dulbecco's Modified Eagle Medium. Monolayers were assigned to one of three experimental groups—(1) control: exposed to culture medium alone; (2) untreated: exposed to 10 3 live C. andersoni oocysts or (3) epidermal growth factor-treated: apically pre-treated with recombinant human epidermal growth factor and then exposed to Cryptosporidium. Oocyst viability, infection with Cryptosporidium, apoptosis, and integrity of tight junctional Zonula–Occludens-1 were assessed. In addition, live Cryptosporidium oocysts were incubated with epidermal growth factor to assess whether epidermal growth factor had cryptosporicidial activity. Cryptosporidium andersoni oocysts infected all human and bovine monolayers, increased nuclear fragmentation, and disrupted Zonula–Occludens-1. Apical epidermal growth factor significantly reduced infection with C. andersoni in all cell lines and inhibited the Cryptosporidium-induced apoptosis and disruption of Zonula–Occludens-1. Epidermal growth factor did not affect oocyst viability.