VSL#3 probiotic preparation has the capacity to hydrolyze gliadin polypeptides responsible for Celiac Sprue probiotics and gluten intolerance

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.

Gluten induces coeliac-like disease in sensitised mice involving IgA, CD71 and transglutaminase 2 interactions that are prevented by probiotics

“…25 Intraluminal therapies of CD are based on the use of ancient variants or genetic modifications of wheat as well as the pretreatment of flour by means of sourdough fermentation using certain lactobacilli with subsequent proteolysis of gluten. 16 Our results showed that oral administration of the S. boulardii KK1 strain in the G þ mice ameliorated the clinical manifestations of coeliac-like disease induced by gluten, decreased the local production of proinflammatory cytokines and downregulated the expression of CD71, which was confined to the basolateral pole of enterocytes. KK1 treatment of the G þ mice increased COX-2 production in the small intestine, which is potentially related to processes that maintain gut mucosa integrity.…”

“…Saccharomyces Boulardii KK1 Strain, a Presumptive Probiotic, Improves Gluten-Induced Enteropathy Because probiotics have been shown to hydrolyse gliadin in vitro, 16 we examined whether presumptive probiotics with established immunoregulatory activity 14,15 have an effect in vivo in the gluten-induced mouse model for CD. The hydrolytic activity of L. paracasei DC412, DC205 or S. boulardii KK1 strains was first verified in vitro by preparing wheat flour fermentation in the presence of these strains.…”

“…Among the tested strains only the KK1 strain exhibited specific proteolytic activity on the 28-kDa a-gliadin (Figure 6a), which is the most toxic fraction in CD. 16 We next investigated whether changes in the microflora of the intestinal lumen by oral delivery of the KK1 strain modulate gluten enteropathy in G þ mice (Figure 6b). Oral administration of the KK1 strain resulted in a decrease in villous Probiotics improve gluten enteropathy C Papista et al atrophy, V:C ratio (Po0.001) and cell accumulation in the lamina propria (Figures 6c and d), whereas delivery of the DC412 strain accelerated the appearance of clinical manifestations such as hair loss (data not shown).…”

“…16 Gliadin was then isolated as described above. Samples were then subjected to western blot analysis as described below using serum isolated from G þ mice as the source of AGA.…”

Designing a Microbial Prolyl Peptidase Delivery System for the Treatment of Celiac Disease

“…Prolyl peptidase activity has already been characterized in several species in the LAB family including, Lactobacillus helveticus, Lactobacillus sakei, Lactobacillus reuteri and Lactococcus lactis (Degraeve et al, 2003;Sanz et al, 2001;Rollan & Font de Valdez, 2001;Xu et al 2002). Furthermore, LABs are capable of detoxifying glutenderived peptides during food processing (Rizzelo et al 2007, De Angelis et al 2006). …”