Transport of beta-lactoglobulin across rabbit ileum in vitro.
Intestinal transepithelial transport constitutes a major limiting step in the transfer of food protein antigens to the blood. This transport was studied in isolated rabbit ileum in Ussing chamber in vitro for the milk protein antigen beta-lactoglobulin (beta-Lg). The transepithelial passage of beta-Lg was measured by enzyme-linked immunosorbent assay (ELISA) and radiolabeled protein transfer and compared with that of the nonmetabolizable marker polyethylene glycol (PEG)-4000. When 1 mg/ml of beta-[14C]Lg or [3H]PEG was added to the mucosal side of the tissue, the total uptake, measured as the transfer of radiolabeled material across the ileum, was significantly higher for beta-Lg than for PEG (5.46 +/- 1.75 vs. 1.43 +/- 0.26 micrograms.h-1.cm-2). Measured by ELISA, 6-9% of the total amount of beta-Lg transported was absorbed in an intact antigenic form. This transport of intact beta-Lg was inhibited by the metabolic inhibitors 50 mM 2-deoxyglucose and 1 mM azide added simultaneously, was reduced by the microtubule assembly inhibitor 0.05 mM colchicine, and was enhanced by 20 mM ammonia, which inhibits lysosomal proteolytic activity. These results indicate that beta-Lg is efficiently absorbed by the intestinal mucosa of adult animals, partly in intact antigenic form and that beta-Lg transport is probably transcellular, as observed for other proteins. The finding that beta-Lg is absorbed in intact antigenic form agrees with other reports implying that beta-Lg is the main factor responsible for milk protein immunoreactivity and intolerance.
Marcon-Genty
D
D
Institut National de la Santé et de la Recherche Médicale, Hopital Saint-Lazare, Paris, France.
Tome
D
D
Kheroua
O
O
Dumontier
A M
AM
Heyman
M
M
Desjeux
J F
JF
D000641:Ammonia / Q000494:pharmacology; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003078:Colchicine / Q000494:pharmacology; D003847:Deoxyglucose / Q000494:pharmacology; D004594:Electrophysiology; D004797:Enzyme-Linked Immunosorbent Assay; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D007082:Ileum / Q000502:physiology*; D066298:In Vitro Techniques; D007408:Intestinal Absorption / Q000187:drug effects; D007413:Intestinal Mucosa / Q000502:physiology*; D007700:Kinetics; D007782:Lactoglobulins / Q000378:metabolism*; D008297:Male; D011092:Polyethylene Glycols / Q000378:metabolism; D011817:Rabbits
D007782:Lactoglobulins; D011092:Polyethylene Glycols; D000641:Ammonia; D003847:Deoxyglucose; D003078:Colchicine