1. Early nutrition and immunity- progress and perspectives1
Early nutrition and immunity- progress and perspectives
Sonja Lang
Katja Bohländer
Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics, Dr. Alexander Haslberger, LVNr

2. Overview Tolerance Role of nutrition Feeding practises2
Tolerance
Role of nutrition
Feeding practises
Role of dendritic cells, lactobacilli
Intestinal colonization
PUFA
LCPUFA
Lipid rafts
Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics, Dr. Alexander Haslberger, LVNr

3. Immunological tolerance3
Lifelong processes
Polarization of Th cells: Th2*, Treg ↑↑
*Recognition of ultra-low antigen dosis (IgE, IgA)
Sterile GIT
Exposure to bacteria at term and after (mother´s skin, breast milk  maturation of infant´s gut
Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics, Dr. Alexander Haslberger, LVNr

4. Nutrition + immunologic development4
Nutrition
…might affect ID during pregnancy, suckling period, introduction of formula and solide foods 
…source of antigens IS must become tolerant
…provides factors, which modulate immune maturation + responses + influences intestinal flora  antigen exposure, immune maturation, immune response
Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics, Dr. Alexander Haslberger, LVNr

5. German Infant Nutritional Intervention Study5
Effects of hydrolysed and standard cow´s milk formula
human milk feeding: ↓ allergic diseases at 1y
Hydrolysed formula: ↓ atopic dermatitis
Extensively hydrolysed formula: - allergy preventive effect
Partially hydrolysed formula: + allergy preventive effect
Keeping pets (dogs!)  atopic diseases ↓
Caesarean section: different gut flora, antibiotics  diarrhoea, allergic sensitization↑
Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics, Dr. Alexander Haslberger, LVNr

6. Dendritic cells + Lactobacilli6
Function of DC:
drive differentiation of naive Th cells into Th1, Th2 or Treg cells
Treg cells: prevention of autoimmunity, allergy
L. reuteri + L. casei prime human DC and drive development of Treg cells by targeting DC-SIGN
Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics, Dr. Alexander Haslberger, LVNr

7. IMMUNOFLORA study 7
„…how early intestinal colonization affects the development of putative Treg cells and clinical allergy in Swedish infants“
Western infants have a delayed acquisition of several gut microbes and a reduced turnover of strains in intestinal flora Exposure ↓, variety ↓ of environmental bacteria
Early food allergy ↔ poor colonization with S. aureus (strong T cell stimulation)
Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics, Dr. Alexander Haslberger, LVNr

8. PUFA  in the intake of saturated fatty acids8
 in the intake of saturated fatty acids
 in the intake of n-6 family of PUFA
Linoleic acid
Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics, Dr. Alexander Haslberger, LVNr

9. N-6 family of PUFA Linoleic Acid Arachidonic acid 9 Prostaglandine
PGE2
Leukotriene
LTB4
Tromboxanes
TXA2
Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics, Dr. Alexander Haslberger, LVNr

10. 4-series leukotrien = mediators of allergic inflammation:10
= mediators of allergic inflammation:
Vascular permeability
Leucocyte chemotaxis
Respiratory burst
Production of inflammatory cytokines
HYPOTHESIS:  intake of linoleic acid  prevalence of atopic disease
Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics, Dr. Alexander Haslberger, LVNr

11. n-3 family of PUFA -Linolenic acid EPA DHA Increased consumption:11
-Linolenic acid
EPA DHA
Increased consumption:
→ incorporation into immune cells
→ decrease the production of prostaglandin E2 and other eicosanoids
Protective towards allergic disease
E.g. n-3 LCPUFA status was lower in cord blood serum from pregnancy of allergic compared with non allergic mothers.
Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics, Dr. Alexander Haslberger, LVNr

12. n-3 family of PUFA Positive results in patients with asthma12
Positive results in patients
with asthma
n-3 LCPUFA intervention:
Stronger impact on fetal and
neonatal Th1/Th2 immune responses compared to immune responses beyond early infancy
Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics, Dr. Alexander Haslberger, LVNr

13. n-3 LCPUFA Influence on T-cell functional responses and signalling13
Influence on T-cell functional responses and signalling
First, prostaglandin E2 influence the activity of DC, differentiation of naive
T-cells and activity of Th1 and Th2 cells
Second mechanism:
Direct alteration of gene expression through modification of transcription factor activity
Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics, Dr. Alexander Haslberger, LVNr

14. n-3 LCPUFA
EPA and DHA give rise to a novel family of eicosanoid-like mediators, called D- and E- resolvins
Inhibition (in vitro):
T-cell proliferation,
Production of IL-2 and IFN-
Surface expression of CD25
Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics, Dr. Alexander Haslberger, LVNr

15. Evaluation of Allergenicity of Genetically Modified Foods
Report of a Joint FAO/WHO Expert Consulation on Allergenicity of Foods Derived from Biotechnology
January 2001
Rome, Italy

16. Introduction 29 May to 2 June 2000 Joint FAO/WHO Geneva, Switzerland
follow-up: 22 to 25 January 2001
Rome, Italy
members: 28 experts and authors of discussion papers

17. Allergenicity Most frequently asked questions
safety of genetically foods
reliable methodology to assess the allergenicity of foods produced by the recombinant DNA technique needed

18. Scope
General consideration of allergenicity of genetically modified foods
Consideration of the decision-tree approach
Specific questions arising in relation to the assessment of allergenicity of genetically modified foods

19. Food Allergies
„Overhwhelming pathological reactions of the body due to intercurrent contact with antigens“ Clemens von Pirquet 1906
IgE-mediated allergy
Cell-mediated allergy
Oral allergy syndrome

20. Decision tree Criteria source of the transferred genetic material,
molecular weight,
sequence homology,
heat and processing stability,
effect of ph and/or gastric juices and
prevalence in foods.

21. Probability of Allergenicity Source of gene allergenic
Yes No
+/ / /-
High Low
Probability of Allergenicity
Likely
Allergenic
Sequence Homology
Target Serum
Screen
Source of gene allergenic
Pepsin Resistance & Animal Models
Specific Serum Screen

22. Post marketing surveillance
Traceability and labelling
Lack of background data
Many confounding food and non-food related factors
Changes in diets over time
Lack of trained experts an infrastructure

23. Other criteria
Level of expressions
Unintended effects

24. Evaluation of Allergenicity of Genetically Modified Foods
Martina Pomper

25. Ines Pree, Immunology and Food, WS 2006
1. Risk assessment and food allergy: the probabilistic model applied to allergens Spanjersberg, M.Q.I., Kruizinga, A.G., Rennen, M.A.J., Houben, G.F., Food Chem Toxicol, 45: (2007)
Ines Pree, Immunology and Food, WS 2006

26. The probabilistic approach in food allergy
Purpose: avoidance of hidden or undeclared allergens
 Risk assessment
Conservative determinstic appraches: worst case value „an allergic reaction cannot be excluded“
Probabilistic approach quantifies health risk asessment by
Hazard identification: situation, symptoms, target organs
Hazard characterization: threshold, minimum dose
Exposure assessment: intake etc.
Ines Pree, Immunology and Food, WS 2006

27. Hazelnut allergens in chocolate - a case study
Risk assessment of three bars, each of a different brand, according to:
Prevalence
Threshold (LOED*)
Consumption pattern
Allergen concentrations
Computer software
*lowest observed eliciting dose
Result:
The allergen was detectable in each bar but at different concentrations.
Ines Pree, Immunology and Food, WS 2006

28. The probabilistic vs. the deterministic approach
The deterministic model does not distiguish between the different degree of contamination.
“An allergic reaction cannot be excluded” is true for all three brands.
The probabilistic model gives more detailed information and avoids overestimation of the risk for the population.
All three brands together: highest mean risk of 0.05%, i.e. less than 500 subjects per million will respond.
The risk for breakfast consumption is higher when compared to lunch. There was a lower risk for women, since men consume more.
Brand 3: the highest risk of 0.004%; less than 40 subjects will respond which reflects a lower contamination of brand 3
Ines Pree, Immunology and Food, WS 2006

29. Ines Pree, Immunology and Food, WS 2006
2. Practical and predictive bioinformatics methods for the identification of potentially cross-reactive protein matches. Goodman, R.E. Mol Nutr Food Res, 50: (2006).
Ines Pree, Immunology and Food, WS 2006

30. Potential allergenicity in GE food
If the protein similar to a known allergen, specific IgE may be cross-reactive (recognition of similar epitopes)
sequence  conformation  cross-reactivity
How to determine potential allergenicity:
Compare amino acid sequences by computer programs
Recruit potentially at-risk individuals (allergic patients)
Perform serum testing, skin prick testing, food challenge.
Ines Pree, Immunology and Food, WS 2006

31. Comparison of amino acid sequences
FASTA and BLAST alignments (used for species homologies) to identify IgE and T cell epitopes?
Since 1990ies: 8 contiguous amino acid matches
In 2001: 6 amino acid matches are too short, too many matches; >35% identity over 80 amino acids is useful
Points of discussion:
Allergen databases are incomplete, mainly lacking minor allergens
Epitopes are poorly defined and the relevance of conformational epitopes is not fully established
Analysis of 3D structures: group proteins into structural families and compare motif recognition patterns
Ines Pree, Immunology and Food, WS 2006

32. Consensus 2005- workshop in Spain
Short matches are not predictive
FASTA and BLAST algorithms are efficient
Structural comparison may be very useful
There are currently no data to change the guidelines (>35% identity over 80 amino acids)
Ines Pree, Immunology and Food, WS 2006

33. Ines Pree, Immunology and Food, WS 2006
Summary/ Conclusion
In risk assessment of food allergens the current precautionary “may contain” labelling is based on the possible presence of an allergen rather than on the assessment of a quantative risk. The quantative expression of risk could avoid unnecessary labelling or recalls.
In the prediction of IgE cross-reactivities in food allergy: structural comparison may be useful. However, there is currently not enough data to change current guidelines (>35% identity over 80 amino acids).
Ines Pree, Immunology and Food, WS 2006

34. Immunity, Inflammation and Allergy In The Gut
Thomas T. MacDonald and Giovanni Monteleone

35. The gut (1) Nutrients get absorbed
Potential to compromise host defense
infection diseases are largely under control
But: gastrointstinal food allergies have increased
 Probably because of the absence of gut infections has upset the balance between the commensal in the gut

36. The gut (2) High active immunsystem
Barrier is a single layer of epithelium
No completely prevent of antigens entering the tissue
Several mechanism how antigens get trough the epithelium
 Immunsystem gets constantly activated

37. Components of the Immunsystem in the gut
Pattern recognition receptors – recognize conserved structures
Severals receptors like TLR, NOD,..
Recognition of TLR ligands increases gut barrier function
Hsp25 and hsp70
CD4+ T cells
Macrophages
Dendritic cells

38. Activation B and T Cells activated  Expression of α4β7 integrin
TLR or NOD activate NFĸB
 Leads to pro-inflammatory gene expression
Chemokine fine-tune the localisation of the tissue

39. Crohn‘s disease (1) Complex genetic disease
Mucosal ulceration, ulcers penetrate into the gut wall
Antigen is not yet identified
Isolated CD 4+ TH1 cells produce large amount of interferon γ
Overexpression of transcriptionfactor T-bet
Macrophages produce large amount of TH1 inducit cytokines
T-cells show resistence to apoptotic signals and have an increased cell cycle

40. Crohn‘s disease (2)
Genes located on the chromosomes 1, 5, 6, 12, 14, 16 and 19
Different polymorphism in the Nod2 gene
 Mutations in the Nod 2 can lead to a decreased ability to kill gut bacteria
OCTN and DGL5 gen
 Important for epithelial permeability
 Disruption leads to inappropriate exposure of the mucosal immunsystem to bacterial products

41. Celiac disease (1)
In some genetically susceptible individuals after ingestion of cereal products
Treated by adherence to a gluten free diet
morphological chances to the mucosa of the upper bowel – long crypts and atrophy of villi

42. Celiac disease (2) 4 components involved
Gluten is prolin and glutamine rich, has negatively charged residues
tTG deaminates glutamin to glutamic acid and produces negatively charged residues
Necessary for efficient binding to HLA-DQ2 and furthermore activation of gluten specific t-cells
Peptides of gliadin activate gut macrophages to produce IL-15 -> increases MICA and arms IEL to kill MICA and epithelial cells

43. Control of inflammation in the gut
T-cells involved in tolerance against commensals
Commensals which crossed the barrier will be phagocytosed without cytokinproduction
 The T-cells die by apoptosis
The epithelial permeability is genetically determined
 Importend factor in the developement of diseases

44. Do They Help to Control Intestinal Inflammation?
Probiotics
Do They Help to Control Intestinal Inflammation?

45. Probiotics
In the maintenance therapy for the inflammating bowel diseases, Crohn’s diseases and ulcerative colitis
Specific molecules modulating defined targets in the gut mucosal and systemic immune system

46. (Active) Ulcerative Colitis
Ulcerative Colitis: Study comparing mesalamine treatment with E.coli Nissle 1917 treatment
relapse rate were not different between groups
E.coli Nissle 1917 is a safe alternative for prevention of relapse in ulcerative colitis
Active Ulcerative Colitis: Trial examining the effectiveness of the fermented milk containing Bifidobacteria strains and Lactobacillus acidophilus.
 Remission was achieved in 4 of 12 patients

47. Pouchitis
Study of the ability of VSL 3 to prevent recurrence of chronical relapsing pouchitis
17 of 20 patients remained in remission
But: Probiotics have failed to demonstrate efficacy

48. Crohn‘s disease
Pioneer study to examine the efficacy of E.coli Nissle 1917 in maintaining remission in Crohn’s diseases
 Groups did not differ in the rate of remission regardless of disease location

49. Conclusions
More effective in preventing relapse of inflammation than suppressing diseases
In active inflammation sufficient data are missing
Genetically engineered bacteria delivering anti-inflammatory cytokines or other biological molecules

50. Allergy, Parasites and the Hygenie Hypothese
Maria Yazdanbakhsh
Peter G. Kremsner
Ronald van Ree

51. Atopy and Allergic diseases
significant increase in pervalence of allergic deseases in the last years
differences in developing and industrial countries
risk faktors such als increased exposure to indoor allergens
childhood infektions shows a negative association with atopy and allergic diseases

52. Atopy enviromental allergen leads to T cell stimulation
release of Cytokines (IL4, IL5, IL13)
raised IgE levels
Increased numbers of eosinophiles and mast cells

53. Hygenie Hypothese High hygenie Vaccination Antibiotics
Limited exposure to pathogens during early childhood
 Can lead to atopies and allergies

54. Helminth infections Stimulates TH2 Immunresponses
 High levels of IgE, eosinophiles and mast cells
People with helminth infections are rarely afflicted by allergic diseases
 TH2 can‘t be the sole factor for allergic attack

55. IgE blocking hypothesis
Highly not specific polyclonal IgE
Unspecific IgEs satures the Fc-receptors on mast cells
The binding site is blocked
degranulation is inhibited
Hypersensitiviy will be immediated

56. Blocking antibodies
Parasites specific IgG4 antibodies inhibit IgE mediated degranulation of effector cells
Possible mechanism of allergen immunotherapy
IL-10 stimulates the IgG4 differentation
Allergic individulas express lower levels of IL-10

57. True or False? The Hygenie Hypothesis for Crohn‘s disease
Bret A. Lashner M.D.
Edward V. Loftus Jr.M.D.

58. Lack of exposures to enteric pathogens makes one susceptible to Crohn‘s disease
Multiple childhood infections and poor
hygenie protects
 Host develops tolerance or immunity to agents that could trigger Crohn‘s disease

59. Crohn‘s disease
2 different studies came to very different conclusions regarding the hygenie hypthesis
The results of one study supports the hygenie hypothesis
Exposure to pathogens in childhood stimulates the immune system
But the other contradicts
Poor hygenie may contribute to the pathogenesis
much work still needs to be done to determine if childhood exposure are truly important to the pathogenesis of Crohn‘s disease

60. The PRODIA study
Ann N Y Acad Sci 1079: (2006)
Probiotics for the Prevention of Beta cell Autoimmunity in Children at Genetic Risk of Type 1 Diabetes
Section of a pancreas of a dog
Source: Gray´s anatomy
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics Markus Hoffmann

61. Diabetes mellitus
Aretaeus of Capadocia: diabaínein „passing through“ or „siphon“
Thomas Willis (1675): mellitus „sweet taste“
Chronic disorder of carbohydrate, fat and protein metabolism caused by
lack or non-functionality of insulin
Hyperglycemia  Polyuria, excessive thirst, polyphagia, weight loss, fatigue, blurred vision, muscle cramps, nausea
Ketoacidosis
Nonketotic hypersomolar coma
Retinal damage
Chronic renal failure
Diabetic neuropathy
Coronary artery disease
Gangrene: „Diabetic foot“
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics Markus Hoffmann

62. Type 1 Diabetes mellitus (T1DM)
A.k.a Insulin-dependent diabetes mellitus (IDDM) or juvenile diabetes
Loss of the insulin-producing β-cells of the pancreas leading to deficiency of insulin
Type 2 Diabetes mellitus (T2DM)
A.k.a. Non insulin-dependent diabetes mellitus (NIDDM) or adult-onset diabetes
Combination fo defective insulin secretion and defective responsiveness to insulin
Type 3 Diabetes mellitus (T3DM)
A.k.a. Gestational Diabetes
Reduced receptivity to insulin of pregnant women due to their hormone status
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics Markus Hoffmann

63. The Islets of Langerhans
Paul Langerhans 1869
One million islets/pancreas
Combined weight 1-1,5 gramms (1-2% of pancreatic mass)
~1000 cells/islet
65–80% β-cells producing Insulin and Amylin
15–20% α-cells producing Glucagon
3–10 % δ-cells producing Somatostatin
1% PP-cells producing pancreatic polypeptide
Porcine Islet of Langerhans
Brightfield Hematoxylin / Immunofluorescence

64. Etiology of T1DM Genetic factors
Prevalence in the general population: 0,2–0,4%
Concordance rate in monozygotic twins: 40-50%
Concordance rate in dizygotic twins and siblings: 5-10%
Genetic susceptibility accounts for ~half of the etiology
Eighteen Loci (IDDM1-IDDM18) identified by positional cloning. All except four have turned out to be statistical artifacts due to underestimation of the sample size required for meaningful statistical power
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics Markus Hoffmann

65. Genetic factors 1. HLA Class II (IDDM1 Locus)
40-50% of the genetic risk
Predisposing haplotypes:
DRB1*0301(DR3)-DQA1*0501-DQB1*0201(DQ2)
DRB1*0401(DR4)-DQA1*0301-DQB1*0302(DQ8)
Heterozygosity DQ2/DQ8
Protective Haplotypes:
DRB1*1501-DQA1*0102DQB1*0602(DQ6)
Horm Res 2005;64:
Molecular mechanism:
Absence of aspartic acid at position 57 of the β chain of the DQ molecule reversing
the electric charge of the peptide binding groove and altering the binding of epitopes
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics Markus Hoffmann

66. Genetic factors 2. INS-VNTR (IDDM2 Locus)
Horm Res 2005;64:
Polymorphism in the 5´flanking region of the insulin gene, consisting of a
variable number of tandem repeats. Either repeats (class I) or repeats (class II). Homozygosity for class I confers a relative risk of 2-3 compared with the dominant protective presence of class II.
Probably due to lower thymic insulin levels  hampered negative selection
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics Markus Hoffmann

67. Genetic factors 3. PTPN22
Protein tyrosine phosphatase, nonreceptor type 22
The nonreceptor tyrosine phospahatase Lyp is specific to lymphocytes and suppressesT-cell activation by dephosphorylating three kinases important to T-cell signaling.
The R620W SNP has also been associated with other autoimmune diseases like
Rheumatoid Arthritis and SLE
R620W maps to a solid 293-kb linkage disequilibrium block containing 6 other known genes and 625 known SNPs.
Horm Res 2005;64:
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics Markus Hoffmann

68. Genetic factors 4. CTLA-4 Cytotoxic T-lymphocyte-associated antigen 4
Horm Res 2005;64:
Encodes T cell receptor that mediates apoptosis in activated T cells
Associations also in Graves disease and autoimmune hypothyroidism
Functional mechanism of the A6230G polymorphism is unknown.
Contribution of the locus is low (relative risk ~ 1,2).

69. Environmental factors
Early exposure to cow milk and gluten
Aberrant development and maturity of the gut immune system
Enterovirus and Rotavirus infections
Vitamin D3 deficiency
Toxins: Rodenticides (Vacor), chemotherapeutic agents (Streptazotocin)
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics Markus Hoffmann

70. T1DM and the gut
In animal models the incidence of T1DM is highest in a low microbial load environment
Diet modifies the development of T1DM in animal models
T cells from human diabetic pancreas show mucosal homing properties
Microbiotic colonization of the newborn´s gut by bacteria may be important for the initial regulation of the developing immune system. Development of
T1DM is associated with intestinal immune activation and enhanced immunity to food antigens.
Probiotics have been shown to support the development and maturity of the gut immune system and could therefore support oral tolerance and
protection against enteral virus infections.
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics Markus Hoffmann

71. Autoantibodies in T1DM 3 major anti-islet autoantibodies:
Type 1a patients: Autoimmune, autoAbs present
Type 1b patients: No evidence of autoimmunity
3 major anti-islet autoantibodies:
Glutamic acid decarboxylase (GADA): Useful marker for confirming etiology in long-standing cases
Tyrosine phosphatase (IA-2A)
Insulin (IAA): The only β-cell specific autoAg, more in DR4
Most children developing T1DM are positive for at least 2 of these markers.
But B cells apparently not strictly required in the autoimmune process.

72. Immune dysregulation in T1DM
There has no primary diabetogenic autoantigen been found yet!
Trigger ??
Bacterial products?
Regulatory Th2 cell anergy?
Immunity, Vol 7,
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics Markus Hoffmann

73. The PRODIA study
Faculty of Health Sciences, Linköping University, Sweden
Pilot study to test feasibility and safety of the protocol. Start in February 2003.
The aim of the main study will be to determine whether the use of probiotics during the first 6 months of life decreases the appearance of β cell autoantibodies in children with genetic risk for Type 1 Diabetes mellitus.
Affected factors involved could be the reduced occurence of
enteral virus infection, enhanced maturation of the gut immune system, reduzed immunization to dietary insulin, or induced immune regulation
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics Markus Hoffmann

74. Study design 1. Selection procedure
Double-blind randomized placebo controlled study
February 2003 to June 2005: Inform all parents to newborn infants at Linköping University Hospital
Informed consent by parents for 1200 children (~60%)
Screen for HLA risk genotypes (presence of risk alleles without protective haplotypes)
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics Markus Hoffmann
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics Markus Hoffmann

75. Study design 2. Treatment
Randomize participants to receive probiotics or placebo
Lactobacillus rhamnosus GG (5 x 109 cfu)
Lactobacillus rhamnosus LC705 (5 x 109 cfu)
Bifidobacterium breve Bbi99 (2 x 108 cfu)
Propionibacterium freudenreichii ssp. Shermani (2 x 10 cfu)
Distributed once a day by parents at home in soluble capsules
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics Markus Hoffmann

76. Study design 3. Readouts Blood samples taken at 6,
12, and 24 months of age
Fecal samples taken at
home at 3 months interval
Microbiological analyses to
detect enterovirus infections
Analysis of compliance
β-cell autoAbs
Monocyte and T cell-derived cytokines
Intracellular signal proteins
(T bet, STAT-4, STAT-6, GATA-3)
Monocyte activation markers upon LPS and THA stimulation
PHA and insulin-dependent T cell responses
Isolation of enterovirus RNA
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics Markus Hoffmann

77. Results 264 children with risk genes among the 1200 participants
1/168 IAA-positive at 6 months of age
1/61 GADA-positive at 24 months of age
1/61 IA-2A positive at 24 months of age
Expected: ~2% prevalence of at least one of the autoAbs at 24 months
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics Markus Hoffmann

78. Conclusions
„The detected number of autoAbs is close to the expected and there is no evidence that the intervention would increase the appearance of beta cell autoimmunity in the children who participate in the study.“
„The PRODIA study protocol seems to be safe and the study protocol
is feasible for the families.“
Mechanistic studies about the development of the immune system and the occurence of eneterovirus infections are ongoing
VO + SE Immunologisch relevante Aspekte von Lebensmittel, Probiotika und Nutrigenomics Markus Hoffmann

79. Wirken Phytoöstrogene immun-modulatorisch?
Präsentation im Rahmen der LV
Immunologisch relevante Aspekte von Lebensmitteln/Nutrigenomics
WS 2006
Nina Zimbelius
Philipp Schatzlmaier

80. Phytoöstrogene/Isoflavone
Soja ist Hauptquelle für Genistein, Daidzein, Equol
Strukturell ähnlich zu 17-Östradiol
Binden an ER, ER
Ziele: Reproduktions-, Immunorgane
Bestandteil von natürlicher Diät (v.a. Asien)
Ebenfalls in modernen Nahrungsergänzungsmitteln (vielfache Dosis)
Bestandteil von Babynahrung (soy-based infant formula)
Potentieller Einfluss auf Immunsystem durch zahlreiche Studien untersucht
Mäuse, Ratten, Menschen
Ergebnisse unvollständig, teils widersprüchlich

81. Humanes Östradiol vs. Soja-Genistein
Estradiol (a human estrogen) and genistein (a phytoestrogen) The similarly-placed hydroxyl groups at both ends of these two molecules allow them to bind to human estrogen receptors.

82. Immun-inhibitorische Effekte von Genistein
Protein Tyrosine Kinase Inhibitor
NO-Produktion in Makrophagen 
T-Zellen-Proliferation durch CD28-AK 
Interleukin-Produktion , TCL tumorizidale Aktivität 
Leukocyten-Adherenz , T-Zellen-Motilität 
Aktivierung von NK-Zellen durch LPS 
 Vorsicht: in vitro Ergebnisse bei teils sehr hohen Konzentrationen (100µM)
historische Diät: < 1µM im Serum bei japanischen Erwachsenen
effiziente Aufnahme bei Kleinkindern: ca. 4 µM
keine Abnormitäten im Erwachsenenalter dokumentiert
allerdings sensibles Alter für Thymusentwicklung

83. in vivo Ergebnisse bei Mäusen und Ratten

84. Effekte von Östrogenen auf T-Genexpression
Ovariektomisierte Mäuse-Babies
Gabe von E2 und Genistein
Genexpression durch DNA arrays untersucht
Gene für Transkription, Apoptose, ZZ beeinflusst
E2 eher , Genistein eher , grosses overlap an Genen
Genistein wirkt auch auf E2-nicht-responsive Gene
Down-Regulierung von CD4-Transkript

85. Ernährungsstudie beim Menschen
23 Männer und 18 Frauen, Hypercholesterinämie, 62 J.
Post-Menopause (Ersatztherapie!)
Drei kontrollierte Diätphasen à 1 Monat:
a) low-fat Kontrollphase
b) high isoflavone soy phase (73mg/d intake)
c) low isoflavone soy phase (10mg/d intake)
Am Anfang und Ende jeder Phase wurden bestimmt:
Körpergewicht, Blutdruck, Lipoproteine/Blutfette
Proteine der akuten Phase im Serum: CRP, SAA
proinflammatorische Cytokine im Serum: IL-6, TNF-

86. Ergebnisse Entzündungswerte

87. Schlussfolgerungen Geschlechts-spezifischer Effekt
IL-6 , immun-stimulatorisch
Negativ: Autoimmunität, CHD
Positiv: Antwort bei Infektionen, Tumor defense
IL-6 reduziert Plasmakonzentrationen von ILGF-1
Geringere Mortalität bei Hormon-abhängigen Tumoren in Asien, auch bei Brustkrebs
Östrogene & Isoflavone wirken antioxidativ
Vermeidung von DNA damage > Anti-cancer effect

88. Referenzen
Cooke, P.S., Selvaraj, V., and Yellayi, S. (2006) Genistein, Estrogen Receptors, and the Acquired Immune Response. J. Nutr. 136:
Jenkins, D.J.A., Kendall, C.W.C., Connelly, P.W., Jackson, C.C., Parker, T., Faulkner, D., and Vidgen, E. (2002) Effects of High- and Low-Isoflavone (Phytoestrogen) Soy Foods on Inflammatory Biomarkers and Proinflammatory Cytokines in Middle-Aged Men and Women. Metabolism 51(7):

89. Nutritional Genomics
genes
nutrients
diet
molecular processes
health

90. Dietary Factors direct and indirect influence
transcriptional, translational and posttranlational control

91. Intestinal Lumen – mucosal immune system
nutritional environment
hormone – independent:
> nutrients
hormone - dependent
gene regulation

92. Nutrigenetic and nutrigenomic effects
nutrigenetic effect:
influence of polymorphisms on altering the response to dietary components
nutrigenomic effect:
ability of different food components to increase or depress gene expression

93. FABPs – fatty acid binding proteins
lipid balance
control of metabolic and inflammatory pathways
modulation of FABP activity
> regulation of lipid-sensitive pathways??

94. Cancer prevention ω-3 Poly Unsaturated Fatty Acids (PUFAs)
> influence on expression of genes
> anti-cancer activity
> downregulation of synthesis and expression
> induction of pro-apoptotic proteins

95. Leptin communicates information of the bodies fat stores
altered levels of leptin > eating disorders (anorexia nervosa)
regulates different genes (SCD1)
> leptin resistance in obese individuals

96. Interleukin 1 genetics, inflammatory mechanisms, and nutrigenetic opportunities to modulate diseases of aging
Kenneth S. Kornman

97. Inflammation healthy person inflammatory mediators no disease disease
genetics
smoking
body mass index
inflammatory mediators
> concentration
nutrition
no disease
disease
complex chronic disease

98. Interleukin 1 IL-1 and TNF-a > early activated
drugs that block their activity
> treatment of rheumatoid arthritis

99. Interleukin 1 gene variations
Interleukin 1 (IL-1) single-nucleotide polymorphisms
IL-1A
IL-1B
IL-1RN
IL-1 cluster
(+4845) or
(-511) or
(+2018) or
haplotype
(-889)
(+3954)
(-31)
VNTR 1 2 2b 3

100. Increased risks haplotype 1 > periodontitis
haplotype > cardiovascular disease
haplotype > gastric cancer

101. Nutrients interact with inflammatory genes
poly unsaturated fatty acids (PUFAs)
> inhibition of secretion of IL-1 and TNF-a
nutrients that alter the oxidation-reduction status of the cell
different effects in individuals with different polymorphisms

102. Conclusion better understanding of polymorphisms
> identify at-risk persons > interventions
screening for bioactive nutrients
problems:
> costs (testing of asymptomatic people)
> primary preventive measures