Receptor-mediated transcytosis of botulinum neurotoxin A through intestinal cell monolayers.
Botulism is mainly acquired by the oral route, and botulinum neurotoxin (BoNT) escapes the gastrointestinal tract by crossing the digestive epithelial barrier prior to gaining access to the nerve endings. Here, we show that biologically active BoNT/A crosses intestinal cell monolayers via a receptor-mediated transcytosis, including a transport inhibition at 4 degrees C and a passage at 37 degrees C in a saturable manner within 30-60 min. BoNT/A passage rate was about 10-fold more efficient through the intestinal crypt cell line m-IC(cl2), than through the carcinoma Caco-2 or T84 cells, and was not increased when BoNT/A was associated with the non-toxic proteins (botulinum complex). Like for neuronal cells, BoNT/A binding to intestinal cells was mediated by the half C-terminal domain as tested by fluorescence-activated cytometry and by transcytosis competition assay. A 'double receptor model' has been proposed in which BoNT/A interacts with gangliosides of GD(1b) and GT(1b) series as well as SV2 protein. Gangliosides of GD(1b) and GT(1b) series and recombinant intravesicular SV2-C domain partially impaired BoNT/A transcytosis, suggesting a putative role of gangliosides and SV2 or a related protein in BoNT/A transcytosis through Caco-2 and m-IC(cl2) cells.
Couesnon
Aurélie
A
Unité des bactéries anaérobies et Toxines, Institut Pasteur, 28 rue du Dr Roux, 75724 Paris cedex, France.
Pereira
Yannick
Y
Popoff
Michel R
MR
D001665:Binding Sites; D001692:Biological Transport / Q000502:physiology; D019274:Botulinum Toxins, Type A / Q000737:chemistry / Q000378:metabolism*; D018938:Caco-2 Cells; D002460:Cell Line; D002469:Cell Separation; D004705:Endocytosis / Q000502:physiology; D004847:Epithelial Cells / Q000378:metabolism; D005434:Flow Cytometry; D005732:Gangliosides / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000378:metabolism*
D005732:Gangliosides; D008562:Membrane Glycoproteins; D020033:Protein Isoforms; D011956:Receptors, Cell Surface; C033851:ganglioside, GD1b; C016481:trisialoganglioside GT1; D019274:Botulinum Toxins, Type A