Immunology, Faculty of Medicine University of Bari, Bari (Italy) Proprietà benefiche di latte, vino e olio extra vergine di oliva
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1 Immunology, Faculty of Medicine University of Bari, Bari (Italy) Proprietà benefiche di latte, vino e olio extra vergine di oliva Professor Emilio Jirillo
2 basketball court (about 400 m 2 ) mucosa-associated lymphoid tissues (MALT) nasal-associated lymphoid tissues (NALT) Peyer s patch (PP) bronchus-associated lymphoid tissues (BALT) isolated lymphoid follicle (ILF) gut-associated lymphoid tissues (GALT) intraepithelial lymphocyte (IEL) genitourinary-associated lymphoid tissues (GUALT) lamina propria (LP)
3 gut-associated lymphoid tissues (GALT) secretory IgA IEL M DC plasma cell T cell B cell M Peyer s patch MLN lamina propria
4 Production of secretory IgA by the gut immune system pigr secretory IgA M PP T cell DC M IL-4, IL-5 IL-5, IL-6 TGF- B cell siga + B cell plasma cell lamina propria inductive tissue effector tissue
5 Milk effects on the immune system
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11 Effects of red grape polyphenols on the immune system in experimental models and humans
12 General structure of flavonoids A 1 O C ' 1' B 3' 6' 4' 5' Non flavonoids compound 5 O C6-C3-C6 Resveratrol 3,4,5 trihidroxy-trans-stilbene fitoalexin Resveratrol OH HO OH
13 Ring skeletons of flavonoids O O O O Flavone O Flavonol OH Flavanol OH O O O O Flavanone OH O Flavanonol O OH O + O Proanthocyanidin OH O Isoflavone OH Anthocyanidin Polymer form of Flavanols
14 Proliferation Migration NO ET-1 I-CAM EDHF MCP-1 V-CAM HO OH Endothelial cells Aggregation Adhesion OH Polyphenols Oxidized LDL Foam cell formation Platelet M
15 IKK- IKK- MAL MyD88 MD-2 MD-2 polyphenols LPS TLR4 LPS LBP LPS scd14 IRAK4 IRAK1 IRAK2 UBC13 TRAF6 TAB2 TAB1 TAK1 MEK1/2 MKK7 MKK3 UBV1A IKK- p38 JNK cytokines, NO p50 I B p65 Proteasome I B NF- B
16 Fermented Grape Marc (FGM) and Intestinal Immunity
17 THE POLYPHENOLS FAMILY Koshu Negroamaro H 2 O ethanol H 2 O ethanol catechin, epicatechin: green flavonol: yellow + catechin, epicatechin: green catechin, epicatechin: green flavonol: yellow + catechin, epicatechin: green
18 Materials for our experiments Koshu Negroamaro Tannat (Japan) (Italy) (Italy)
19 The manufacturing process of FGM Grape marc Supermasscolloider (Ultra-fine friction grinder) Grape marc : H2O = 1 : 1 sterilization at 121 C, 15 min incubate 1% lactic acid for 1 day bacteria media (Lactobacillus plantarum) add. 5% (v/v) fermentation for 4 days sample (FGM) freeze-drying
20 Inflammatory bowel disease (IBD) inflammatory area ulcerative colitis colon Crohn s disease gastrointestinal tract damage shallow intestinal wall deep into the layers of intestinal wall cytokine response Th2 dominant Th1 dominant
21 Induction of colitis 5% dextran sodium sulfate (DSS) FGM (10, 30, or 100 mg/kg in water, p.o.) BALB/c mice Normal group Control group FGM-treated group days 5% DSS p.o water water each of FGMs body weight colon length TNF- or IL-1 production in homogenized colon
22 Control Koshu Koshu Negroamaro Tannat percentage of body weight loss (%) Effect of FGM on body weight loss fermentation (100 mg/kg)
23 Control Koshu Koshu Negroamaro Tannat percentage of shortening of colorectum (%) Effect of FGM on length of colon * fermentation (100 mg/kg)
24 Comparison of length of colon in FGM (Koshu)-treated mice Normal Control fermented Koshu (mg/kg)
25 Normal Control Normal Control pg/ml pg/ml Effect of FGM (Koshu) on inflammatory cytokines in homogenized colon TNF-α IL-1β * ** * * fermented Koshu (mg/kg) fermented Koshu (mg/kg)
26 Fermented Grape Marc and effects on human PBMCs
27 Evaluation of cytokine release from human PBMCs stimulated by N-FGM and K-FGM
28 Intracellular content of cytokines in human PBMCs N-FGM/ water N-FGM/ethanol K- FGM/water K- FGM/ethanol IL-12 (monocytes) IL-10 (lymphocytes) u.n. 9% 17% u.n. u.n. 20% u.n. 6% IL-10 (monocytes) 6% u.n. 7% u.n. TNF- (monocytes) 8% u.n. u.n. 7% u.n. undetectable % increase vs untreated cells
29 Intranuclear staining of FoxP3 in Treg cells Ab anti-cd4 CD4+ CD25+ Ab anti-cd25 FoxP3 Ab anti- FoxP3
30 unstimulated Positive control (PMA) FGM-stimulated cells
31 FoxP3 expression N/w Negroamaro/water N/et Negroamaro/ethanol K/w Koshu/water K/et Koshu/ethanol FoxP3 n.s. n.s.
32 Percent induction of Treg cells by N-FGM and K-FGM
33 GrB release by human PBMCs stimulated with N-FGM and K-FGM
34 Detection of Granzyme B in supernatant of lymphocytes culture
35 Effects of FGM on granulocytes and monocytes
36 Effects of Extra Virgin Olive Oil on the Immune-Mediated Inflammatory Responses: Potential Clinical Applications
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38 In vitro polyphenol effects on the immune system from obese people
39 Determination of absolute numbers of peripheral blood mononuclear cells (PBMCs)
40 H unstim CD3+ Ob unstim CD3+ H CD3+ 1ug Ob CD3+ 1 ug H CD3+ 3 ug Ob CD3+ 3 ug H CD3+ 5ug Ob CD3+ 5 H unstimulated vs Ob H 1ug/ml vs Ob 1 H 3 ug/ml vs Ob 3 H5 ug/ml vs Ob 5 ug/ml
41 # # # * H unstim CD4+ Ob unstim CD4+ H CD4+ 1ug Ob CD4+ 1 ug H CD4+ 3ug Ob CD4+ 3ug H CD4+ 5ug Ob CD4+ 5 ug # p<0.001 H unstimulated vs Ob unstimulated # p<0.001 H 1 ug/ml vs Ob 1 ug/ml # p<0.001 H 3 ug/ml vs Ob 3 ug/ml * p<0.05 H 5 ug/ml vs Ob 5 ug/ml
42 H unstim CD8+ Ob unstim CD8+ H CD8+ 1ug Ob CD8+ 1 ug H CD8+ 3ug Ob CD8+ 3 ug H CD8+ 5ug Ob CD8+ 5 H unstimulated vs Ob H 1ug/ml vs Ob 1 H 3 ug/ml vs Ob 3 H5 ug/ml vs Ob 5 ug/ml
43 * 0 H unstim CD19+ Ob unstim CD19+ H CD19+ 1ug Ob CD19+ 1 ug H CD19+ 3ug Ob CD19+ 3 ug H CD19+ 5ug Ob CD19+ 5 ug # p<0.001 H unstimulated vs Ob p< H 1 ug/ml vs Ob 1ug/ml *p<0.05 H 3 ug/ml vs Ob 3 ug/ml
44 # # H unstim CD16CD56+ On unstim CD H CD16CD56+ 1ug Ob CD16+CD56+ 1 ug H CD16CD56+ 3ug Ob CD16+CD56+ 3 ug H CD16CD56+ 5ug Ob CD16+CD56+ H unstimulated vs Ob H 1 ug/ml vs Ob 1 ug/ml #p<0.001 H 3 ug/ml vs Ob 3 ug/ml #p<0.001 H 5 ug/ml vs Ob 5 ug/ml
45 Polyphenols do not influence the increased absolute numbers of obese PBMCs
46 Effects of polyphenols on Th1 related-cytokines
47 IL-2 release
48 IFN- release
49 In vitro treatment with polyphenols did not affect the release of both IL-2 and IFN-
50 Effects of polyphenols on Th2 related-cytokines
51 IL-4 release
52 In vitro treatment with polyphenols did not affect the release of IL-4 The enhanced release of Th1 and Th2-related cytokines in the presence of PMA suggests a condition of immune hyperactivation in obese people as also indicated by the increase in PBMC absolute numbers
53 Effects of polyphenols on Treg/Th21 axis
54 The inflammatory /anti-inflammatory pathway T regulatory cells Production of IL-10, the antiinflammatory pathway Inhibition of Th17 cells (IL-17A-F), the inflammatory pathway IL-21 operates as an inductor of Th17
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57 Spontaneous release of IL-17 in obese people was absent, thus depending on both IL-10 production and IL-21-induced suppression by polyphenols.
58 Spontaneous release of IL-10 is moderately increased by polyphenols Release of IL-21, an inducer of Th17 cells, was abrogated in the presence of polyphenols while they significantly enhanced production of this cytokine by normal PBMC
59 In vitro effects of polyphenols on pro-inflammatory cytokine release
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65 Reduced release of IL-1 and IL-6 from obese PBMCs by polyphenols in comparison to the normal counterpart reflects the anti-inflammatory pathway above described
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