Lactobacillus Paracasei CBA L74 prevents entrance of undigested - - PowerPoint PPT Presentation

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Dec 02, 2023 47 likes •208 views

Lactobacillus Paracasei CBA L74 prevents entrance of undigested gliadin peptides and rotavirus in Caco-2 cells Naples September 23-25 2014 Merlin Nanayakkara, Marco Sarno, Roberto Nigro Riccardo Troncone, Salvatore Auricchio Department of

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Lactobacillus Paracasei CBA L74 prevents entrance of undigested gliadin peptides and rotavirus in Caco-2 cells

Naples September 23-25 2014 Merlin Nanayakkara, Marco Sarno, Riccardo Troncone, Salvatore Auricchio Roberto Nigro Department of Chemistry European Laboratory for the Investigation of Food Induced Diseases Department of Traslational Medical Science University of Napoli, Federico II,Italy and M.Vittoria Barone Vittori Buccirossi Department of Chemistry University of Napoli, Federico II,Italy Andrea Budelli Heinz.

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Celiac disease

Is a multifactorial disease caused by gluten ingestion in genetically susceptible subjects. The damage in the celiac intestine is mediated by an immune response both adaptive and innate, causing crypts hypertrofia and villus atrophy Diagnosis: antibodies anti TTG and anti endomisium Biopsy Therapy: Life long total abstinence from gluten containing food Other enviromental factors: Drugs (INF-alpha) and viral infections

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Some gliadin peptides are resistant to digestive enzimes

(Mamone G. et al J Chromatography B, 855(2):236-241, 2007 P31-43 P57-68

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Gliadin peptides dual activity

LGQQQPFPPQQPY QLQPFPQPQLPY

Innate immunty PEPTIDE “T-CELL IMMUNOGENIC” PEPTIDES

Prototype P57-68 Prototype P31-43 QLQPFPQPQLPY Some gliadin peptides that are deamidated by tissue transglutaminase bind to typical CD HLA, DQ2 and/or DQ8 molecules, and induce an adaptive Th1 pro-inflammatory response (ie P56-68). Other gliadin peptides are able to initiate a response involving innate immunity independently from HLA interactions (ie P31-43) .

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Gliadin peptides enters into the cells by endocytosis Interaction of ‘toxic’ and ‘immunogenic’ A‐gliadin peptides with a membrane‐mimetic environment

J of Molecular Recognition Vilasi s et al 2009 Caputo I. et al. Biochim Biophys Acta. 2010

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LP CBA L74 effect on gliadin peptides entrance is concentration dependent

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Supernatant of LP CBA L74 effect on gliadin peptides entrance

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Supernatant of LP CBA L74 intereferes with endocytosis of dextran

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Cereals fermented with LP CBA L74 intereferes withgliadin peptides endocytosis

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Effect of LP CBA L74 supernatant on rotavirus (RV) entrance in RV‐infected CaCo‐2 cells

CTRL RV

No Infection RV Infection rescence intensity

6 8 10 12 14 16 18

Sup Sup + RV

LP CBA L74 LP CBA L74 + RV Infection Fluoresce

2 4 6 RV Infected LP CBA L74 +RV Infection

Figure 1

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Effect of LP CBA L74 supernatant on reactive

xygen species (ROS) in RV‐infected Caco‐2 cells

90 10 20 30 40 50 60 70 80 CTRL RV LP sup RV+LP sup

AIF/tot proteins

* p<.001 vs CTRL # p<.001 vs RV

Figure 2

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ELFID Lab.

Prof. Salvatore Auricchio

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Gliadin peptide P31-43 is similar to HRS

(Hepatocyte growth factor-regulated tyrosine kinase substrate)

Hrs is a key protein for the regulation of Hrs is a key protein for the regulation of endocytic maturation endocytic maturation

Barone et al PloS One 2010, 2011 Clatrin binding domain

p31-43

HRS has many binding partners. P31 HRS has many binding partners. P31-

43 is similar to a region of HRS needed for its correct localization to the

43 is similar to a region of HRS needed for its correct localization to the endocytic vesicles endocytic vesicles

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P31-43 competes with HRS localisation

Barone et al PloS One 2010

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P31‐43

Delays endocytic vesicles dinamics Delays maturation of early vesicles

.9 .8 .7 .6 .5 .4 .3 .2 .1 Vesicles speed (µm in 10 min) 30 min. 3h 3h 30 min. P31-43-liss P56-68-liss

EEA1/P31-43 30min EEA1/P31-43 3h LAMP 2/P31-43 3h

Caco2 cells Biposies

Control CD

EEA1/P31-43 3 h

P31‐43

Prolongs EGFR activation

EEA1/P31-43 24 h Min at 37 C: 0 20’ 40’ 90’ 90’

WB: α α α α‐EGFR α α α α‐Tyr(P) ip: α-EGFR Ab

90’ 90’

PTG P31‐43 EGFR EGFR . Time lapse. CaCo2 cells treated with p31- 43 and P57-68 lissaminated. Vesicles containing P31-43 liss are slower that p57- 68 containing vesicles

Barone et al GUT 2007 Gastroenterology 2007 Plos One 2010

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Gliadin peptides can delay endocytic maturation and increse recycling vesicles

IL-15 trans-presented

Proliferation of epithelial cells Innate immunity M.V. Barone et al