Presentation on theme: "Protein hydrolysates: Do they really work ?"— Presentation transcript:
1 Protein hydrolysates: Do they really work ? Vincent C Biourge, DVM PhD Dipl ACVN & ECVCNRoyal Canin, Centre de Recherche, Aimargues
2 Adverse Reactions to Food Dermatologic: pruritus, alopecia,otitis, ..Gastrointestinal: vomiting, abd. discomfortRespiratory: asthma, rhinitis, ...General: headache, arthritis, ...Food AllergyFood intoleranceImmune mediated(IgE, cell mediated)Non immune mediated(Lactose, chocolate, bioamines,additives, …)Probably the main cause (Olivry T, J Vet derm 2010)
3 Adverse Reactions to Food DiagnosisChallengeDr Thierry Ximenes (France)Double blindedClin. signsElimination dietClinical signs4 to 12 WeeksNovel ingredients+ Serum IgE+ Skin testImprovement of the clinical signs
4 Elimination diets Definition Gold standard Diets composed of ingredients to which the dog has not been exposed.Dietary historyHome madeStarch sources: Rice, Potato, TapiocaProtein sources: Lamb, Horse, Fish, Turkey, Venison, ..Essential fatty acids, Minerals and VitaminsCommercialLamb & Rice, Fish & Potatoes, Venison & PotatoesOver the counterNot hypoallergenic !Intact protein sources !SustainabilityFish, Rabbit, Duck, Venison, …Raditic et al, JAPAN 2010
5 A new strategy to treat adverse reaction to food Protein hydrolysates2001A new strategy to treat adverse reaction to foodEnzymaticHydrolysisproteinpolypeptides
6 Benefits of hydrolysate Easier to digestShort half-life in the lumenSmaller peptidesMolecular weight < 16 KdCave, 2006
7 Protein hydrolysates Do they really work ? What is the background ? Should molecular weight be as low as possible ?Are they trully hypoallergenic ?Can they be qualified as allergen free ?Are their efficacies substantiated by clinical studies ?
8 Elisa immunoreactivity of soy protein depending of its source BackgroundElisa immunoreactivity of soy protein depending of its sourceLaMartin.comAdverse reaction to soy in milk replacerPoor appetite and growthMalabsorption/maldigestion – DiarrheaSoy antibodiesVillus atrophyLallès, 1995Soy flourSoy concentrateSoy hydrolyzate
9 Protein hydrolysate based milk replacer. BackgroundProtein hydrolysate based milk replacer.Baby allergy to cow’s milkVomiting diarrheaAtopic dermatitis/ UrticariaAsthma/RhinitisBabies at risk for allergy Risk of cow milk allergy
10 Molecular weight Dalton Arbitrary unit of atomic mass Sir John Dalton, Founder of the atomic theory1 Dalton (d) = Weight of 1/12 nuclide of 12CgrammeGlycin = 75 d - Tryptophan = 204 dCasein = d or KdMeasurement: electrophoresis -chromatographyHuman serum albumin = 69 Kd
11 Molecular weightMean MWMeanMaxIngredients Vs dietsCave N, 2006
12 Molecular weight Most common food allergens Glycoproteins Protein with glucide moiete? Lipids (Bacterial glycolipids)? CarbohydratesWater - solubleHeat and acid resistantsMolecular weight 10 to 40 KdSmall enough to pass the intestinal wallLarge enough to induce immune reactionAntigens in selected foodin manCave N, 2006Looringh van Beeck FA, 2009Sampson HA, 1993
13 Highly reduced allergy Molecular weightMolecularweightLess common allergy> 40 kDaMost commonkDaReduced allergy3 – 10 kDaGuilford GW ,1996Serra et al, 2006Highly reduced allergy1– 3 kDaER: But we are targeting no allergies. Studies of children have established that only amino-acid-based and extensively hydrolysed formulas might be considered truly hypoallergenic (Hill, 2007) and Diets lower than 1kDa will ensure the greatest chance of reducing allergic reaction (Cave 2006)Olivry T, 2010Cave N, 2006No allergy< 1 kDa
14 Molecular weight Antigenicity Epitope Few Many Close in 3D Hidden Gastroscopic score diameterOlson ME et al 2000Epitope
16 Molecular weight Digestibility Soy source and in vitro N digestibility Soy source and aa ileal digestibility in calfsLallès, 1995Royal Canin, 2004Cave NJ, Marks SL. Evaluation of the immunogenicity of dietary proteins in cats and the influence of the canning process. AJVR 2004; 10,
17 Molecular weight Cut-off « The number of IgE binding sites on the allergen, their location, and the tertiary shape of the protein are probably more important than the molecular weight »S.F. Hefle, 1996Source of protein (Casein Vs Soy)Process - type of hydrolysis -exposure of epitopesProblem measuring MW on finished product
20 Immunogenicity Skin test Puigdemont et al, 2006 % of reduction of the wheal areas for soy hydrolysedversus native soy proteinSoy protein concentrationDog1 g/ml10 g/ml100 g/ml1T253,845,742,01T353,554,652.62T295,618,444,62T681,96186,43T259,758,437,43T44780,157,6Mean SD65,3 19,153,0 20,453,4 17,7Puigdemont et al, 2006Control showed no wheal on both challenges
21 Immunogenicity Clinical reactions Dogs Native soy protein Hydrolysed specific IgE8204VomitsDiarrhoea (1)PruritusNR+++8212Diarrhoea (2)8216++Control and 3 other sensitised dogs showed no adverse reactionsPuigdemont et al ,2006
22 Immunogenicity Clinical reactions Jackson et al, 2003 Cutaneous Score(Max 35*3*3)Score max = 315: 3 types de lésions (erythema, excoriation, infection) gradé de 0 à 3.Pas de réaction avec le corn starchNo corn and starch dietCorn starchCornSoySoy hydrolysate diet200 mg/kg bw14 maltese x Beagle dogs with known clinical hypersensitivity to soy and cornJackson et al, 2003
23 Immunogenicity Stomach reactivity Gastroscopic score diameter Olson ME et al 2000
24 Immunogenicity Conclusion Hydrolyzed proteins are less antigenic than intact proteins.Hydrolyzed proteins are not anallergenic except if hydrolyzed to single or few amino acids.High digestibility is an important factor in reducing antigenicity.
25 Clinical trials Shown efficacy Managing adverse reaction to food. Diagnosis of adverse reaction to food.Inflammatory bowel disease.Exocrine pancreatic insufficiency.
26 Adverse reaction to food Clinical reactionsCutaneousClinicalScoreNo corn and starch dietCorn starchCornSoySoy hydrolysate diet200 mg/kg bw14 maltese x Beagle dogs with known clinical hypersensitivity to soy and corn3/14 dogs increased scores on hydrolyzed diet.Jackson et al, 2003
27 Adverse reaction to food Evaluation on dogs with demonstrated adverse reactions to food12 dogs ( breeds, sex, food allergy )Controlled allergy (no clinical signs)Clinical 0 and 2 months (CADESI)Owners requested to report:Pruritus, abnormal behavior of the dogDigestive tolerancePalatibilityNone of the 12 dogs relapsedPerfect toleranceJ. Fontaine, CNVSPA 2001
28 Diagnosis of ARF Challenge Clin. signs Soy hydrolysate diet 2 dermatology specialty praticesJ Fontaine (Brussels, B), M Vroom (Oisterwijk, NL)Inclusion in the study:Suspicion of skin hypersensitivityJ Nutr2002;134:2062S-2064SChallenge8 WeeksClin. signsSoy hydrolysate dietClinical signsRecovery challenge + Adverse Reaction to Food (ARF)2. Marked improvement challenge + ARF + atopy3. Little or no improvement other elimination diet Recovery4. Little or no improvement other elimination diet no improvement ? atopy No corticotherapyVC Biourge, J Fontaine, MW Vroom, 2004
29 20 16 22 Diagnosis of ARF Atopy Adverse Reaction to Food 60 dogs included31 M-3 MC - 13 F - 13FSAge 4.5±0.4 yrs (3 mo – 11 yrs)26 breedsGerman Sheperd (10), Bouledogue Français (2), Bouvier des Flandres (2), Boxer(5), English Cocker Spaniel (2), Golden Retriever (4), Jack Russel (2), Labrador (4), Shar Pei (3), Shi Tsu (2), WHWT (5), …Duration of the clinical signs2.6±0.4 yrs (3 weeks – 10 yrs)201622AtopyAdverse Reactionto Food+ 2 cases excluded
30 Adverse reaction to food Diagnosis of ARFAdverse reaction to food20 dogs9 M-1 MC - 6 F – 4 FSAge 3.8±0.6 yrs (6 mo – 9yrs)No more pruritusNo or very mild clinical signs left.Challenge +18/20 responded to soy hydrolysate dietRabbit and RiceHomemade soy diet***
31 Diagnosis of ARF Before After Golden Retriever Male, 4.5 yrs, 29.6 kg Golden retriver, mâle, 29,6 kg :J 0 :Prurit chronique intense et généralisé (initialement à l’abdomen), avec une évolution en alopécie, lichénification-hyperpigmentation et corticosensible seulement à l’apparition des symptômes de l’allergie.J 30 :Reduction du prurit et des lésions cutanées spécialement au niveau de l’abdomen et des yeux.J 60 :Prurit résiduel.Les lésions cutanées sont acceptables.Confirmation de l’allergie alimentaire : réapparition des symptômes suite à la réintroduction de l’ancienne alimentation.Golden Retriever Male, 4.5 yrs, 29.6 kgGeneralized intense prurit with lichenification, hyperpigmentationBeforeAfter
32 Adverse reaction to food & Atopy Diagnosis of ARFAdverse reaction to food & Atopy16 dogs8 M-1 MC - 2 F – 5 FSAge 5.7±0.7 yrs (3 mo – 11yrs)Pruritus marketly improvedMild to moderate clinical signs left.Challenge +All dogs responded to the soy hydrolysate diet.***
33 Diagnosis of ARF Atopy 22 dogs 13 M-1 MC - 5 F – 4 FS Age 4.5±0.6 yrs (1.2 – 11yrs)No or little improvement of pruritus, clinical signs.No response to other elimination diets and + to skin test
34 ARF: response to the hydrolysate Diagnosis of ARFARF: response to the hydrolysate94.4 % of ARF dogs responded to the soy hydrolysate based diet
35 Diagnosis of ARF 15 20 47 10 11 38 Other study Loeffler et al., Vet Derm 2006;17:Chicken hydrolysate diet Vs homade diets181 dogs17 dogs excluded35 allergic skin diseasePoultry hydrolyzate (109)Homade (72)Excluded b/c did not receive exclusively the food PH 13 – HD 4, did not eat the food: PH 5 – HD 4, needed glucocorticoïds PH 1 – HD 3, Diarrhea PH 1 – HD 1, unrelated reason = 215204710No significant difference1138ARFAtopyARFAtopyExcluded: 27Excluded: 13
36 1 dog showed severe signs when fed hydrolyzed chicken Veterinary Dermatology,2010,21:12 dogs selected and divided In 2 groupsDuring PH1, 25 of 26 dogs showed an increased CADESIscore after chicken meat once daily for 2 weeks. Twelvesubjects, falling into classes 3 and 4, were selected forPH2 of the study. Prior to starting PH2, one dog devel-oped unrelated problems and was substituted with aclass 2 dog. Gastrointestinal disturbances were not recor-ded during PH1 or PH2.The average CADESI score (LSMEANS) calculatedduring the entire study was not inﬂuenced by the group.Dogs displayed the most prominent clinical responsewhile eating whole chicken, whereas no signiﬁcantdifferences were detected between PH2.1 and PH2.2(PH1: versus PH2.1: versus PH2.2:2.69,P < 0.001; Figure 2).1 dog showed severe signs when fed hydrolyzed chicken
37 Diagnosis of ARF Cats After Before Dr Aranda Golden retriver, mâle, 29,6 kg :J 0 :Prurit chronique intense et généralisé (initialement à l’abdomen), avec une évolution en alopécie, lichénification-hyperpigmentation et corticosensible seulement à l’apparition des symptômes de l’allergie.J 30 :Reduction du prurit et des lésions cutanées spécialement au niveau de l’abdomen et des yeux.J 60 :Prurit résiduel.Les lésions cutanées sont acceptables.Confirmation de l’allergie alimentaire : réapparition des symptômes suite à la réintroduction de l’ancienne alimentation.AfterBeforeDr Aranda
38 ARF: response to the hydrolysate Diagnosis of ARFARF: response to the hydrolysate87.5 % of ARF cats responded to the soy hydrolysate based dietMW Vroom, C. Swinnen, A clinical study of a soy protein isolate hydrolysate diet, in dogs and cats with adverse reactions to food. Proc. of Voorjaarsdagen
40 Food responsive chronic diarrhea Dogs:26 dogs over a 2 yr-period.24/26 dogs with IBD4 IF, 10 SF, 8 IM, 4 CM.Age: 4.3 ± 3.3 yrs ( yrs)Weight 23 ± 12 kg ( kg)Duration of the clinical signs:1 to 36 months before presentation.Treatments before inclusion:Antibiotics (7), Metaclopramide (6), cimitidine (6) , Prednisolone (4), sulfazalazine (3).Diets before inclusion:Low residue intestinal diets (7), novel protein diets including homemade (12), other diets (7).Mandigers et al., 2010ObjectivesTo compare the response of dogs with chronic diarrhea on soy hydrolysate Vs intestinal diet.
41 Food responsive chronic diarrhea Clinical signs after 2 monthsWeight gain*11%12.5%No more clinical signs 23/26 dogs3 last dogs improved but vomiting and diarrhea persisted
42 Food responsive chronic diarrhea Follow-up after median 6 months, range 3-15 mo.15/16 test dogs – 6/7 control dogs67%13%87 % of dogs on hydrolyzate diet remained free of clinical signs on follow-up ! The signs were minor in the remaining 2 dogs.
43 Food responsive chronic diarrhea Similar findings by other authors.Marks SL, Laflamme D, McCandlish A. Dietary trial using a commercial hypoallergenic diet containing hydrolyzed protein for dogs with IBD. Vet Ther 2002; 3:Similar finding in cats8 catsChronic diarrhea (4-36 Mo)1 Colitis, 2 gastritis6 IBDSoy hydrolyzate based dietResolution of clinical signs within 4-8 dMedian weigh gain 0.75 kg within 2 Mo11Mo follow-up 6/8 cats free of clinical signs
44 Exocrine pancreatic Insufficiency J Nutr2002;134:2166S-2068SGerman ShepherdsEPI and skin diseaseGI signs controlled within 7 dWeight gain with 2 monthsSkin within 3 months
45 Protein hydrolysates Conclusion Hydrolyzed proteins are less antigenic than intact proteins.Molecular weight (except if extremly low (<1Kd) is a poor predictor of protein immunogenicity.Hydrolyzed protein based diet are not anallergenic.Hydrolyzed proteins are sustainable.Clinical studies to support benefits in:Diagnosis and management of ARFIdiopathic chronic diarrhea - IBDExocrine pancreatic insufficiency? Perianal fistula.