Lactobacillus Paracasei CBA L74 prevents entrance of ... fileEffect of LP CBA L74 supernatant on...
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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.
Transcript of Lactobacillus Paracasei CBA L74 prevents entrance of ... fileEffect of LP CBA L74 supernatant on...
Lactobacillus Paracasei CBA L74 prevents
entrance of undigested gliadin peptides
and rotavirus
in Caco-2 cellsNaples September 23-25 2014
Merlin Nanayakkara, Marco Sarno,Riccardo Troncone, Salvatore Auricchio
Roberto NigroDepartment of Chemistry
European Laboratory for the Investigation ofFood Induced Diseases
Department of Traslational Medical ScienceUniversity of Napoli, Federico II,Italy
and M.Vittoria BaroneVittori Buccirossi
Department of Chemistry University of Napoli, Federico II,Italy
Andrea BudelliHeinz.
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 endomisiumBiopsy
Therapy: Life long total abstinence from gluten containing food
Other enviromental factors: Drugs (INF-alpha) and viral infections
Some gliadin peptides are
resistant to digestive enzimes
(Mamone G. et al J Chromatography B, 855(2):236-241, 2007
P31-43
P57-68
Gliadin peptides dual activity
LGQQQPFPPQQPY QLQPFPQPQLPY
Innate immunty PEPTIDE “T-CELL IMMUNOGENIC” PEPTIDES
Prototype P57-68Prototype P31-43
QLQPFPQPQLPY
Some gliadin peptides that are deamidated by tissuetransglutaminase bind to typical CD HLA, DQ2 and/or DQ8molecules, and induce an adaptive Th1 pro-inflammatory response(ie P56-68).Other gliadin peptides are able to initiate a response involvinginnate immunity independently from HLA interactions (ie P31-43) .
Gliadin peptides enters into the cells by endocytosisInteraction of ‘toxic’ and ‘immunogenic’
A‐gliadin peptides with a membrane‐mimetic
environment
J of Molecular Recognition Vilasi s et al 2009Caputo I. et al. Biochim Biophys Acta. 2010
LP CBA L74 effect on gliadin peptides entrance is concentration dependent
Supernatant of LP CBA L74 effect on gliadin peptides entrance
Supernatant of LP CBA L74 intereferes with endocytosis of dextran
Cereals fermented with LP CBA L74 intereferes withgliadin peptides endocytosis
Effect of LP CBA L74 supernatant on rotavirus
(RV) entrance in RV‐infected CaCo‐2 cells
CTRL RV
No Infection
RV Infection
Flu
ores
cenc
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tens
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6
8
10
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N4
Sup Sup + RV
LP CBA L74
LP CBA L74+
RV Infection
Flu
ores
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tens
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0
2
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RV Infected LP CBA L74 +RV
Infection
Figure 1
Effect of LP CBA L74 supernatant on reactive
oxygen species (ROS) in RV‐infected Caco‐2 cells
90
0
10
20
30
40
50
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CTRL RV LP sup RV+LP sup
AIF
/tot
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tein
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* p<.001 vs CTRL
# p<.001 vs RV Figure 2
ELFID Lab.Prof. Salvatore Auricchio
Gliadin peptide P31-43 is similar to HRS
(Hepatocyte growth factor-regulated tyrosine kinase substrate)
Hrs is a key protein for the regulation ofHrs is a key protein for the regulation of
endocytic maturationendocytic maturation
Barone et al PloS One 2010, 2011
Clatrin bindingdomainp31-43
HRS has many binding partners. P31HRS 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 vesiclesendocytic vesicles
P31-43 competes with HRS localisation
Barone et al PloS One 2010
P31‐43
Delays endocytic vesicles dinamicsDelays maturation of early vesicles
.9
.8
.7
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0
Ve
sicl
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spe
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(µ
m
in 1
0 m
in)
30 min. 3h 3h30 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:
αααα‐EGFR
αααα‐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 2007Gastroenterology 2007Plos One 2010
Gliadin peptides can delayendocytic maturation and increse recycling vesicles
IL-15 trans-presented
Proliferation of epithelial cellsInnate immunity
M.V. Barone et al
