Immunoregulatory activity of dietary omega3 polyunsaturated fatty acids

Immunoregulatory activity of dietary omega-3 polyunsaturated fatty acids in T cells Wooki Kim, Ph. D. Dept of Food Sci & Biotech, Kyung Hee University

Immunity A resistance of an organism against infection or disease. Antigen Exogenous: food, pathogen, pollen, etc Endogenous: self-peptides, damaged tissues, tumors Activation vs Regulation Hostdefense Selfimmunity

Chronic Inflammation: Body Under a Fire

Immune systems CD 4 T cell Innate immunity : Fast, No memory Adoptive immunity : Slow, Memory

CD 4+ T-cells control adaptive immune system Zhu et al. Blood. 2008. 112, 1557 -69

CD 4 T cell: a Control Tower Th 1 Virus, Cancer Chronic inflammation Th 2 Bacteria, Peptide Allergy

Again, Balance Matters Chronic inflammation Pathogenesis Th 1 Th 2 Activation Regulation

Factors affecting CD 4 T cell function Genetic background Age, Gender Pathogen, Hygienity Gut flora Habits: Alcohol, Smoking, Exercise Diets: Vitamins, Ribonucleotides, Phytochemical polyphenols, Lipids

n-3 Polyunsaturated Fatty Acids: Potential Immunoregulator Fish oil Corn oil

Old Greenland Story Disease Incidence in Greenland Expected incidence Diabetes mellitus 1 9 Psoriasis 2 40 Bronchial asthma 1 25 Multiple sclerosis 0 7 Total 4 81 Kromann, N. et al. Acta Med Scand. 1980

Human Clinical Trials Disease Number of studies Rheumatoid arthritis 14 Crohn’s disease 3 Ulcerative colitis 4 Psoriasis 2 Reviewed by Calder, P. C. Lipids. 2001

Hypothesis 1 Dietary n-3 PUFA inhibit Th 1 development in vivo.

CD 4 T cell Adoptive Transfer DO 11. 10 CD 4 T cell transgenic mice OVA+ CFA (Th 1) CD 4 T cell n-3 vs n-6 Normal mice

Flowcytometry Cell size Granulity Surface markers Cell count

Donor cell (%) FO decreased lymphocyte accumulation * n-6 n-3 Zhang et al. J Nutr. 2006. 136: 2391 -2398

Cell Trafficking vs Proliferation Cell Injection

Cell Trafficking vs Proliferation n-3?

Cell Trafficking vs Proliferation Cell Injection

Cell Trafficking vs Proliferation Immunization

Cell Trafficking vs Proliferation n-3?

Cell Trafficking vs Proliferation n-3?

Cell division analysis by CFSE

CD 4 T cell Adoptive Transfer OVA+ CFA (Th 1) Normal mice

FO suppressed Th 1 cell division in vivo OVA n-6 No OVA n-3 CFSE Zhang et al. J Nutr. 2006. 136: 2391 -2398

Summary 1 Dietary fish oil (n-3 PUFA) suppressed Th 1 cell division. Chronic inflammation Pathogenesis Th 1 Th 2 Activation Regulation

Question What is the molecular mechanisms by which CD 4 T cell function is affected by n-3 PUFA?

Proposed Action of n-3 PUFA Wooki Kim et al. Prosta, Leukotri & Essent Fatty Acids. 2009

Lipid rafts: a putative target of n-3 PUFA Cholesterol/ sphingolipid rich Liquid ordered Heterogeneity Signaling platform Small: 5 -200 nm Highly dynamic DHA Fan et al. 2004. J Immunol. 173, 6151 -6160 Pike. 2006. J Lipid Res. 47: 1597 -1598

TCR proximal signaling APC MHC Ag TCR/CD 3 complex T-cell L A T Lck Jury et al. Sem Cell Dev Biol. 2007. 18: 608 -615

TCR proximal signaling n-3 PUFA L A P T L A T PLC γ 1 P Grb 2 SLP 76 Gads PLC γ 1 PIP 2 F-actin Lck PKCθ P CARMA 1 NF-k. B signaling F-actin IP 3 DAG Zap 70 Ca 2+ signaling PKCθ CARMA 1 Jury et al. Sem Cell Dev Biol. 2007. 18: 608 -615

Hypothesis 2 n-3 PUFA promote lipid raft formation at the immune synapse, thereby altering signaling molecule recruitment and activation in CD 4 T-cells

Laurdan: A lipid raft indicator in living cells ↑ GP Lipid rafts ↓ GP Non-raft Gaus et al, Mol. Memb. Biol. 23. 41 -48. 2006

Immunological synapse: a docking station Magnification 40 x IS Wooki Kim et al. J. Immunol. 2008.

n-3 PUFA enhance lipid raft formation at the IS * Wooki Kim et al. J. Immunol. 2008.

Protein patching at the IS Magnification 40 x non-patching CD 4+ T-cell 1 o anti-LAT patching 2 o anti-Ig. GAlexa 568 APC % conjugate with patching = (# cells with patching) / (# cells examined) Wooki Kim et al. J. Immunol. 2008.

n-3 PUFA suppress relocalization of signaling proteins at the IS % Conjugate with patching 80 * 70 60 * * 50 40 30 20 10 0 PKCθ CARMA 1 LAT n-6 WT PLCγ 1 F-actin n-3 fat-1 Wooki Kim et al. J. Immunol. 2008.

Phosphorylated protein patching at the IS patching 1 o anti-p-PLCγ 1 2 o anti-Ig. GAlexa 568 % conjugate with patching = (# cells with patching) / (# cells examined) Wooki Kim et al. J. Immunol. 2008.

n-3 PUFA suppress PLC 1 activation at the IS *P=0. 034 n-6 n-3 Wooki Kim et al. J. Immunol. 2008.

Suppressed Proliferation by n-3 PUFA n-6 n-3 Wooki Kim et al. J. Immunol. 2008.

n-3 PUFA enhance lipid raft formation to suppress signaling and proliferation L A T Lck P P F-actin P Grb 2 SLP 76 Gads PLC γ 1 Zap 70 T-cell proliferation PKCθ CARMA 1

Summary Balance CD 4 T cell activation vs regulation. Th 1 vs Th 2 differentiation. Hyperactivation of Th 1 cells results in chronic inflammatory disease. n-3 PUFA suppress Th 1 differentiation. n-3 PUFA increase lipid raft formation, thereby suppress T cell signaling cascade.

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