An In Vitro Model of Infection of Human Biliary Epithelial Cells by<i>Cryptosporidium parvum</i>

Verdon, Renaud; Keusch, Gerald T.; Tzipori, Saul; Grubman, Shelley A.; Jefferson, Douglas M.; Ward, Honorine

doi:10.1086/593695

Cited by 32 publications

(34 citation statements)

References 10 publications

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“…3 Development of human biliary epithelial cell lines now allows in vitro modeling of biliary tract infection by Cryptosporidium. 4 Confirming and extending the observations of Verdon et al, 4 Chen et al 5 report in the present issue of HEPATOLOGY that Cryptosporidium only invades the luminal surface of cultured human biliary epithelial cells and fails to attach to the basal surface. This implies the existence of apical receptors for infective sporozoites, which can be blocked by antibody as the mechanism of immunologic clearance of infection and which could conceivably be blocked by introduced therapeutic agents.…”

supporting

confidence: 84%

New perspectives on the pathogenesis of Cryptosporidium biliary disease

Owen

1998

Hepatology

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supporting

confidence: 84%

New perspectives on the pathogenesis of Cryptosporidium biliary disease

Owen

1998

Hepatology

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“…Our work confirms a single recent study that demonstrated that cultured human biliary epithelial cells can be infected by C. parvum. 26 However, our work substantially extended this work in several ways. First, we demon- HEPATOLOGY Vol.…”

Section: Discussionmentioning

confidence: 83%

Cryptosporidium parvum is cytopathic for cultured human biliary epithelia via an apoptotic mechanism

Chen¹,

Levine²,

Tietz³

et al. 1998

Hepatology

101

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While the clinical features of sclerosing cholangitis secondary to opportunistic infections of the biliary tree in patients with acquired immunodeficiency syndrome (AIDS) are well known, the mechanisms by which microbial pathogens such as Cryptosporidium parvum associated with this syndrome actually cause disease are obscure. We established an in vitro model of biliary cryptosporidiosis employing a human biliary epithelial cell line. Using morphological and biochemical techniques, we examined the interaction of C. parvum with cultured human cholangiocytes. When the apical plasma membrane of polarized, confluent monolayers of human biliary epithelial cells was exposed to C. parvum oocysts that had been excysted in vitro, sporozoites attached to and invaded the cells in a time-, dose-, temperature-, and pH-dependent manner. The infectious process was both plasma membrane domain-and cell-specific, because no attachment or invasion occurred when the basolateral membrane of cholangiocytes was exposed to the parasite, or when a human hepatocyte cell line (HepG2) was used. Time-lapse video microscopy and scanning electron microscopy (SEM) showed that sporozoite attachment was rapid, involved extensive cholangiocyte membrane ruffling, and culminated in parasite penetration into a tight-fitting vacuole formed by invagination of the plasma membrane similar to those found in naturally occurring infection in vivo. Transmission electron microscopy (TEM) showed that C. parvum organisms formed parasitophorus vacuoles and were able to undergo a complete reproductive cycle, forming both asexual and sexual reproductive stages. Unexpectedly, direct cytopathic effects were noted in infected monolayers, with widespread programmed cell death (i.e., apoptosis) of biliary epithelial cells as assessed both morphologically and biochemically beginning within hours after exposure to the organism. The novel finding of specific cytopathic invasion of biliary epithelia by C. parvum may be relevant to the pathogenesis and possible therapy of the secondary sclerosing cholangitis seen in AIDS patients with biliary cryptosporidiosis. (HEPATOLOGY 1998;28:906-913.)

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“…Four IBEC lines were derived by transduction of normal BALB/c intrahepatic BEC with SV40 large T antigen. 22,24 No attempt was made to subclone any of the lines; thus, they most likely contain heterogeneous populations of BEC. IBEC were extensively compared with freshly isolated BEC from BALB/c mice without bile duct ligation to avoid contamination with hyperplastic cholangiolar epithelial cells.…”

Section: Discussionmentioning

confidence: 99%

“…15 Collectively, these phenotypic changes indicate that BEC may be targets for either class I MHC-restricted CD8 or class II MHC-restricted CD4 CTL bearing FAS-ligand 16 and might function as ''professional'' antigen-presenting cells (APC). 17 However, the capacity to study the immunopathogenesis of NSDC and the immunologic function of isolated [18][19][20][21] or immortalized [22][23][24] human BEC remains limited.…”

mentioning

confidence: 99%