title: Bacillus thuringiensis insecticidal Cry1ab toxin does not affect the membrane integrity of the mammalian intestinal epithelial cells: An in vitro study.
abstract: The mammalian intestinal epithelium has been found, based on in vivo experiments, to be resistant to insecticidal Cry toxins, which are derived from Bacillus thuringiensis and fatally damage insect midgut cells. Thus, the toxins are commonly used as a genetic resource in insect-resistant transgenic plants for feed. However, Cry toxins bind to the cellular brush border membrane vesicle (BBMV) of mammalian intestinal cells. In this study, we investigated the affinity of Cry1Ab toxin, a lepidopteran-specific Cry1-type toxin, to the cellular BBMV of two mammalian intestinal cells as well as the effect of the toxin on the membrane potential of three mammalian intestinal cells compared to its effects on the silkworm midgut cell. We found that Cry1Ab toxin did bind to the bovine and porcine BBMV, but far more weakly than it did to the silkworm midgut BBMV. Furthermore, although the silkworm midgut cells developed severe membrane potential changes within 1 h following the toxin treatment at a final concentration of 2 mug/ml, no such membraneous changes were observed on the bovine, porcine, and human intestinal cells. The present in vitro results suggest that, although Cry1Ab toxin may bind weakly or nonspecifically to certain BBMV components in the mammalian intestinal cell, it does not damage the cell's membrane integrity, thus exerting no subsequent adverse effects on the cell.
authors.affiliation: National Institute of Animal Health, Tsukuba, Ibaraki 305-0856, Japan. shimadan@affrc.go.jp
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Shimada
authors.forename: Kazuhisa
authors.initials: K
authors.lastname: Miyamoto
authors.forename: Khozo
authors.initials: K
authors.lastname: Kanda
authors.forename: Hideo
authors.initials: H
authors.lastname: Murata
mesh_terms: D000818:Animals; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000378:metabolism*; D001427:Bacterial Toxins / Q000378:metabolism*; D012831:Bombyx; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D004731:Endotoxins / Q000378:metabolism*; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D006460:Hemolysin Proteins; D006801:Humans; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000166:cytology*; D007476:Ionophores / Q000378:metabolism; D008564:Membrane Potentials / Q000502:physiology; D013552:Swine; D014634:Valinomycin / Q000378:metabolism
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; D007476:Ionophores; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D014634:Valinomycin