Use of ancient wheat crops for the diet of non-celiac ... · Use of ancient wheat crops for the...
Transcript of Use of ancient wheat crops for the diet of non-celiac ... · Use of ancient wheat crops for the...
Use of ancient wheat crops for the diet
of non-celiac gluten sensitive patients
9th PROBIOTICS, PREBIOTICS & NEW FOODS, NUTRACEUTICALS AND BOTANICALS for Nutrition &
Human and Microbiota Health Roma, 10/12 Settembre 2017
Giuseppe Mazzarella
Institute of Food Sciences-CNR - Avellino
Biomarkers
YES
YES
NO
Gluten Related
Disorders
Pathogenesis
Autoimmune Allergic
Not Autoimmune
Not allergic
(Innate immunity?)
Celiac DiseaseGluten
Ataxia
Dermatitis
herpetiformis
Wheat
allergy
Gluten
sensitivity
Respiratory
Allergy
Food
AllegyWDEIA
Contact
UrticariaSymptomitic Silent Potential
Proposed New Classification of Gluten Related Disorders
Adaptive and + Innate Immunity
Frequency Intestinal Extra-intestinal
Very Common Bloating Lack of wellbeing
Abdominal pain Tiredness
Common Diarrhea Headache
Epigastric pain Anxiety
Nausea Foggy mind
Aerophagia Numbness
GER Joint/muscle pain
Aphtous stomatitis Skin rash/dermatitis
Alternating bowel habits Weight loss
Constipation Anemia
Loss of balance
Undetermined Hematochezia Depression
Anal fissures Rhinitis/asthma
Weight increase
Interstitial cystitis
Ingrown hairs
Oligo or polimenorrhea
Sensory symptoms
Disturbed sleep pattern
Hallucinations
Mood swings
Autism
Schizophrenia
The clinical manifestations of Non-Celiac Gluten Sensitivity (NCGS).
DIAGNOSIS
CD: serology ( anti-TG2) and histology ( villous atrophy)
WA: wheat specific IgE antibodies
NCGS: absence of specific diagnostic markers
To date, is mainly a “diagnosis of exclusion”
Pathogenesis of Celiac Disease (CD)
Maria I. Vazquez-Roque et al. Gastroenterology. 2013
Pathogenesis NCGS innate response?
Sapone A et al. Int Arch Allergy Immunol. 2010
Sapone A et al. BMC 2011
Duodenal IHC CD3+ IELs
40 X
A= Gluten Sensitive
B= Controls
C= CD Active
A B
C
Sapone A. et alt, Int. Arch. of Allergy and Immunology 2009
A small increased of
CD3+ intraepithelial
lymphocytes (IELs)
could be detected in
some NCGS patients,
compared to controls,
Research is intense to find
wheat varieties with absent or
low toxicity to be implemented
in new strategies for treatment
and prevention of CD. Among
candidates, there are ancient,
diploid wheat species for their
low capability to activate
intestinal T cell responses in
CD patients
The genealogy of cultivated members of the Triticum
family
Values reported in the literature within and among wheat types for celiac reactivity
Molberg and others 2005; Pizzuti and others 2006; Vincentini and others 2007, 2009; van den Broeck and others 2010a, 2010b
Absence of immunotoxic 33-mer peptide in Triticum Monococcum Einkorn
0
500
1000
1500
2000
2500
3000
3500
0 50 100 150 200 250
ug/ml PT
pg/
ml I
FN-g
PT-Sag TG
PT-Monlis TG
PT-ID-331 TG
Treatment with gliadin peptides from two monococcumaccessions induces γIFN production from all intestinal
gliadin-specific T-cell lines.
Dose-response curves
0
20
40
60
80
100
120
140
CD
25
+ ce
lls/m
m2
lam
ina
pro
pri
a
**
*
*
0
10
20
30
40
50
60
CD
3+
ce
lls/m
m e
pit
elu
im
***
T. aestivum wheat giadin
T. monococcummonlis gliadin
T. monococcumID-331 gliadin
T. aestivum wheat giadin
T. monococcummonlis gliadin
T. monococcumID-331 gliadin
**
Lamina propria CD25+ cells IEL CD3+ cells
IMMUNOHISTOCHEMISTRY
Challenge with gliadin peptides from two monococcum accessions induced increase of IEL and LP CD25 cells in treated CD biopsies.
P TG PT- monlis PT- ID331 Medium
Medium PT-ID331 PT-Monlis
Gliadins from monococcum wheat Monlis but not those from monococcum wheat ID331 induced IL-15 expression in intestinal mucosa
from treated CD patients.
Our data show that the monococcum lines Monlis and ID331 activate the CD T cell response and suggest that
these lines are toxic for celiac patients. However, ID331 is likely to be less effective in inducing CD because of its
inability to activate the innate immune pathways.
Gianfrani C et al. Am J Clin Nutr 2012;96:1339–45
SUMMARY
Gliadin proteins
Pepsin (2h)
Chymotrypsin (4h)
Pepsin (30 min)
Trypsin, Chymotrypsin, Elastase, Carboxypeptidase
(1h)
Brush Border Membrane enzymes
(BBM) (2h)
Deamindation by tTGase (2h)
T-cell and biopsy assay
RP-HPLC analysis
Collected fraction
Chymotrypsin (16h)
nLC-MS/MS
Pepsin (30 min)
nLC-MS/MS
Immunassay Proteomics
Gliadinidentification
Data base search
Data base search
Identification of resistant peptides
Immunogenity evaluation
PC GD-BBM
Trypsin, Chymotrypsin, Elastase,
Carboxypeptidase (1h)
Brush Border Membrane enzymes
(BBM) (2h)
GD-BBM
Pt#CD301012
0
200
400
600
800
1000
1200
Inte
rfero
n-g
(pg
/ml)
PC GD-BBM I331-PC I331 GI-BBM Monlis PC Monlis GI-BBM
Sag
0
20
40
60
80
100
120
140
Sag-PC Sag GD-BBM ID331-PC ID331 GD-BBM
CD
25
+ ce
lls\m
m²
lam
ina
pro
pri
a
*
*
*A
p<0,005
p<0,003
p<0,05
0
2
4
6
8
10
12
14
Sag-PC Sag GD-BBM ID331-PC ID331 GD-BBM
CD
3+C
D2
5+
acti
vate
d T
ce
lls
*
*
*
p<0,005
p<0,005
p<0,001
Marked reduction of specific T-cell response to monococcum gliadin after the GD-BBM digestion
Marked reduction of T cell activation response to monococcum gliadin after the GD-BBM digestion
Sagittario
ID331
Monlis
w a/b g
MS-based analysis showed that several
Triticum monococcum peptides, including
known T-cell epitopes, were degraded
during the gastrointestinal treatment,
whereas many of Triticum aestivum gliadin
survived the gastrointestinal digestion.
SUMMARY
Mazzarella G et al. Mol Nutr Food Res. 2015 Sep;59(9):1844-54
Therefore our data confirm that monocoque grain is not a safe grain for celiac disease, but could
be a cereal candidate to represent a major food alternative for patients with NCGS and / or for
the prevention of celiac disease in subjects at risk for familiarity .
Dose response of IL-8 release by monocyte-
derived DCs stimulated with purified ATI .
LPS served as positive control
Non-gluten α -Amylase/Trypsin Inhibitor (ATI) family
are strong activators of innate immunity.
Junker Y et al J Exp Med2012;209(13):2395-408
ATI bioactivity determination in ancient and modern cereals
Victor F. Zevallos et al. Gastroenterology 2017;152:1100–1113
Triticum Monococcum as a healthy food
The properties of MonLis einkorn compared with a variety of common wheat:
•higher protein content;
•higher content of ash and trace elements (iron, zinc, magnesium, phosphorus, potassium etc.);
•low presence of saturated fatty acids;
•lower starch content and better digestibility;
•higher presence of antioxidants and yellow pigmentation (carotenoids, including beta-carotene, the precursor to vitamin A);
•higher content of tocols (vitamin E). These facts are relevant to the functional activities of cells and are efficient antioxidants
Therefore, these data suggest their potentiality as functional foods
Carmen GianfraniGianfranco MamoneVera Rotondi AufieroISA-CNR, Avellino
Salvatore AuricchioRiccardo Troncone Dept Pediatrics, and ELFIDUniv Federico II, Naples
Acknowledgments
Nicola GiardulloGaetano IaquintoU.O. Gastroenterolgy, HospMoscati, Avellino
Alessio FasanoMassachusetts General Hospital Harvard Medical School
Thanks for your attention