2007 Annual Report 1a.Objectives (from AD-416) 1) To identify the mechanisms of local type 1 and type 2 cytokine-dependent alterations in gut function in the colon and in unaffected areas in the small intestine.. 2) Determine the impact of specific nutritional deficiencies alone (Vitamin E and Selenium) on baseline levels of cytokines and cytokine receptor expression and on gut function.. 3)Determine the impact of specific nutritional deficiencies (Vitamin E and Selenium) on immune-induced alterations in gut function. 1b.Approach (from AD-416) Experiments will compare responses in Wild Type mice or mice deficient in the signaling factors Stat4 or Stat6 after treatment with agents known to elevate Th1 (inflammation using trinitrobenzene sulfonic acid) or Th2 cytokines (enteric helminth infection). Specific nutritional deficiencies that impact these responses and affect gut function will be examined; treatment groups will include. 1)vehicle,. 2)TNBS,. 3)infection,. 4)VE and Se deficient diet). Molecular (real time RT-PCR) will be used to determine diet-induced alterations in cytokine profiles and histological evaluation will be performed to identify modifications in infiltrating cells as a result of the diet deficiencies. Laser capture microscopy (LCM) will be used to localize cytokine receptors to specific cell types including epithelial mucosa, smooth muscle and nerves. Preliminary data indicate that diets deficient in both VE and Se impair the ability of the intestine to clear enteric nematodes and affect intestinal function. Function is defined as changes in epithelial cell secretion and absorption in vitro (Epithelial cell transport in Ussing chambers) and smooth muscle contraction in vitro (Isometric smooth muscle contraction). Evaluation of histological changes and assessment of cytokine and cytokine receptor expression will be determined routinely using LCM and tissue cytokines using RT-PCR. 3.Progress Report This research is part of a collaborative venture with scientists at the Department of Medicine and the Mucosal Biology Research Center, University of Maryland School of Medicine, Baltimore, Maryland that received a competitive grant renewal this year for an additional five years under NIH grant RO1-AI/DK49316. The lead scientist presented components of the work on selenium deficient diets and immune function at the 8th International Congress of Veterinary Immunology in Ouro Preto, Brazil, and a plenary presentation at the 21st World Association for the Advancement of Veterinary Parasitology in Gent, Belgium this summer. 4.Accomplishments Selenium Deficiency Reduces the Local Oxidative Response to Tissue Parasites in the Intestine: The local tissue response to parasitic larvae of Heligmosomoides polygyrus in the small intestine is dependent on alternatively activated macrophages that contain and destroy the worm. Selenium deficiency blocks this host protective response through interference with enzymes that increase the production of oxygen radicals. This is the first demonstration that these cells contribute to oxidative stress to control infectious agents. This observation is important because these cells are also associated with allergic disease, and suggest that diets deficient in antioxidants can contribute to the severity of disease. This mechanism has not been systematically examined and points to a strategy to control allergic responses through diet. Conducted as part of the National Program for Human Nutrition-107 Component 6, Relationship between Diet, Genetics, Lifestyle, and the Prevention of Obesity and Disease. Isolated Parasite Products Modulate Immune Responses: The inherent nature of parasite interaction with the host is to down regulate immunity in order to enhance survival. This logically predicts that parasite products down regulate host immune function. Two proteins were isolated from the large roundworm, Ascaris suum, that in their native form turn off antigen-presenting cell function to interfere with host immunity. These products also affect responses to non-parasite proteins at mucosal surfaces, including the lung and intestines, and can be useful in studies that target these sites to reduce the severity of inflammatory responses. The products can work in tandem with dietary conditions that reduce inflammation in both chronic and acute responses to infection or metabolic stress. Conducted as part of the National Program for Human Nutrition-107 Component 6, Relationship between Diet, Genetics, Lifestyle, and the Prevention of Obesity and Disease. 5.Significant Activities that Support Special Target Populations None. 6.Technology Transfer Number of patent granted 2 Number of non-peer reviewed presentations and proceedings 6 Review Publications Erazo, A., Kutchukhidze, N., Leung, M., Guarnieri-Christ, A.P.,Urban Jr,J.F., Curotto De Lafailli, M.A., Laifaille, J.J. 2007. Unique maturation program of the IgE response in vivo. Immunity. 26(2):191-203. Zaiss, D.M., Yang, L., Shah, P.R., Kobie, J.J., Urban, J.F., Mosmann, T.R. 2006. Amphiregulin-a TH2 cytokine enhancing resistance to nematodes. Science. 314(5806):1746. McConchie, B.W., Norris, H.H., Bundoc, V.G., Trivedi, S., Boesen, A., Urban Jr, J.F., Keane-Myers, A.M. 2006. Ascaris suum-derived products suppress mucosal allergic inflammation in an interleukin-10-independent manner via interference with dendritic cell function. Infection and Immunity. 74(12):6632-6641.