title: VSL#3 probiotic preparation has the capacity to hydrolyze gliadin polypeptides responsible for Celiac Sprue.
abstract: The native structure and distribution of gliadin epitopes responsible for Celiac Sprue (CS) may be influenced by cereal food processing. This work was aimed at showing the capacity of probiotic VSL#3 to decrease the toxicity of wheat flour during long-time fermentation. VSL#3 (10(9) cfu/ml) hydrolyzed completely the alpha2-gliadin-derived epitopes 62-75 and 33-mer (750 ppm). Two-dimensional electrophoresis, immunological (R5 antibody) and mass spectrometry analyses showed an almost complete degradation of gliadins during long-time fermentation of wheat flour by VSL#3. Gliadins non-hydrolyzed during fermentation by VSL#3 were subjected to peptic-tryptic (PT) digestion and analyzed by CapLC-ESI-Q-ToF-MS (Capillary Liquid Chromatography-Electrospray Ionization-Quadrupole-Time of Flight-Mass Spectrometry). Search for several epitopes showed the only presence of alpha2-gliadin-fragment 62-75 at a very low concentration (sub-ppm range). Compared to IEC-6 cells exposed to intact gliadins extracted from the chemically acidified dough (control), VSL#3 pre-digested gliadins caused a less pronounced reorganization of the intracellular F-actin which was mirrored by an attenuated effect on intestinal mucosa permeability. The release of zonulin from intestinal epithelial cells treated with gliadins was considerably lower when digested with VSL#3. Agglutination test on K 562 (S) cells showed that the PT-digest of wheat flour treated with VSL#3 increased the Minimal Agglutinating Activity of ca. 100 times. Wheat proteins were extracted from doughs and subjected to PT digestion. Compared to PT-digest from chemically acidified dough, celiac jejunal biopsies exposed to the PT-digest from the dough fermented by VSL#3 did not show an increase of the infiltration of CD3(+) intraepithelial lymphocytes. Proteolytic activity by probiotic VSL#3 may have an importance during food processing to produce pre-digested and tolerated gliadins for increasing the palatability of gluten-free products.
authors.affiliation: Department of Plant Protection and Applied Microbiology, University of Bari, 70126 Bari, Italy.
authors.forename: Maria
authors.initials: M
authors.lastname: De Angelis
authors.forename: Carlo G
authors.initials: CG
authors.lastname: Rizzello
authors.forename: Alessio
authors.initials: A
authors.lastname: Fasano
authors.forename: Maria G
authors.initials: MG
authors.lastname: Clemente
authors.forename: Claudio
authors.initials: C
authors.lastname: De Simone
authors.forename: Marco
authors.initials: M
authors.lastname: Silano
authors.forename: Massimo
authors.initials: M
authors.lastname: De Vincenzi
authors.forename: Ilario
authors.initials: I
authors.lastname: Losito
authors.forename: Marco
authors.initials: M
authors.lastname: Gobbetti
mesh_terms: D000199:Actins / Q000378:metabolism; D000372:Agglutination Tests; D000818:Animals; D017252:CD3 Complex / Q000276:immunology; D002446:Celiac Disease / Q000378:metabolism*; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D002772:Cholera Toxin / Q000378:metabolism; D017097:Electric Impedance; D005260:Female; D005285:Fermentation; D005903:Gliadin / Q000032:analysis / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D006242:Haptoglobins; D006801:Humans; D006868:Hydrolysis / Q000187:drug effects; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D020014:K562 Cells; D051379:Mice; D008807:Mice, Inbred BALB C; D010447:Peptide Hydrolases / Q000378:metabolism; D010539:Permeability / Q000187:drug effects; D019936:Probiotics / Q000494:pharmacology*; D011498:Protein Precursors; D051381:Rats
chemical_list: D000199:Actins; D017252:CD3 Complex; D006242:Haptoglobins; D011498:Protein Precursors; C408043:zonulin; D005903:Gliadin; D002772:Cholera Toxin; D010447:Peptide Hydrolases