Larvae-induced plasma membrane wounds and glycoprotein deposition are insufficient for Trichinella spiralis invasion of epithelial cells.
Trichinella spiralis L1 larvae infect susceptible hosts by invading epithelial cells that line the small intestine. Invasion also occurs in vitro when larvae are inoculated into cultures of epithelial cells from several different animal species. To further investigate the mechanism of invasion, we studied the interaction of larvae with the rat epithelial cell line IEC-6. Larvae did not invade IEC-6 cells, but did cause the cells to take up parasite glycoproteins. Glycoprotein bearing cells remained viable and were detectable in monolayers for as long as 24 h, suggesting that the glycoproteins were not toxic for cells. Immunofluorescence revealed that parasite glycoproteins localized in the nuclei, mitochondria and cytoplasm and we found evidence for selection of certain molecules between nuclear and cytoplasmic compartments. Using fluorescent dextrans as fluid phase markers we found 17-38% of the cells in inoculated monolayers were engorged with dextran and that dextran was free in the cytoplasm. Increased dextran uptake was not lethal, required the presence of activated larvae, and was often associated with uptake of parasite glycoproteins. These observations suggest that larvae caused plasma membrane wounds. Our results showed that neither delivery of glycoproteins nor mechanical wounding is sufficient to allow entry of the parasite into resistant epithelial cells. Because both invasion-resistant and susceptible epithelial cells undergo non-lethal wounding, we propose that larvae-induced injury to epithelial cells may result in release of cell-specific mediators that signal larva to invade a particular cell line or, alternatively, to ignore it.
Butcher
B A
BA
James A. Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA. bab26@cornell.edu
Gagliardo
L F
LF
ManWarren
T
T
Appleton
J A
JA
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000469:parasitology* / Q000648:ultrastructure; D003911:Dextrans / Q000378:metabolism; D004705:Endocytosis; D004847:Epithelial Cells / Q000469:parasitology*; D005434:Flow Cytometry; D005453:Fluorescence; D006023:Glycoproteins / Q000378:metabolism*; D015801:Helminth Proteins / Q000378:metabolism; D006601:Hexoses / Q000378:metabolism; D007814:Larva / Q000378:metabolism / Q000472:pathogenicity; D018613:Microscopy, Confocal; D051381:Rats; D013347:Subcellular Fractions; D017160:Trichinella spiralis / Q000254:growth & development / Q000472:pathogenicity*
D003911:Dextrans; D006023:Glycoproteins; D015801:Helminth Proteins; D006601:Hexoses; C009616:tyvelose