Progress 10/01/09 to 09/30/13
Outputs
Target Audience: The target audience includes academic researchers in the US and internationally, poultry industry scientists, veterinarians, and government scientists. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? This project provided training for two post-doctorals (Schusser and Manzanilla) and 3 graduate students (Cornax, Iseri, Kurzbard). How have the results been disseminated to communities of interest? Peer-reviewed Journal Publications and scientific seminars. What do you plan to do during the next reporting period to accomplish the goals? Continue to determine the energetic and essential amino acid costs of developing, maintaining and using the immune system in chickens.
Impacts
What was accomplished under these goals?
Objective 1. Identify and characterize genes and their relationships to disease resistance in poultry with an emphasis on the major histocompatibility complex as well as other genes encoding alloantigens, communication molecules and their receptors and other candidate systems. The scavenger receptors MARCO and CD36 were found to be highly expressed in unactivated macrophages and mediated phagocytosis without triggering an inflammatory phenotype. Surprisingly, chicken macrophages were found to highly express T1R1/T1R3 (umami taste receptor), which was found to detect foreign peptides. Objective 2. Identify and characterize environmental, dietary and physiologic factors that modulate immune system development, optimal immune function and immune system related disease resistance and welfare in poultry genetic stocks. Nutrients that have primarily regulatory functions (Vitamin A. D and essential fatty acids) were found to have greater effects on development of the immune system than nutrients that serve as anabolic precursors (amino acids, energy, minerals). Vitamin A deficiency during development diminished B lymphocyte maturation and the breadth of the antibody repertoire. The mechanism by which the immunemodulatory nutrients lutein and n-3 PUFA affect inflammation were explored. Both lutein and n-3 PUFA were found to blunt LPS-induced PPARα and RXRα downregulation. Dietary n-3 PUFA blunted the increase in IL-1 mRNA due to LPS in a PPAR-dependent manner. The effects of lutein appear to have an epigenetic component that allows this immunomodulation and continue after withdrawal for at least one year. This mechanism exists in both commercial broilers (Cobb) and layers (HyLine). The innate and adaptive immune systems must have appropriate levels of nutrients in order to maintain leukocyte numbers and effector proteins in order to facilitate their response. To determine the quantity of nutrients needed for the systemic compartment of the immune system, we monitored the aggregate changes in the mass and amino acid content of leukocytes and effector proteins at maintenance health) and during a response to E. coli injected iv into adult laying hens. During the early response (4-24 hr), leukocytes (except heterophils) were redistributed from blood into tissue pools, especially the spleen. Specific IgM and IgY increased 7d post-injection compared to baseline). As the response progressed, leukocytes in the bone marrow and spleen increased markedly (P<0.05). The data demonstrated a re-distribution of leukocytes from blood to tissue during the early phase and an increase in the concentration of leukocytes and effector proteins during the late response. During the acute phase following E. coli injection the total lysine content of leukocytes decreased (except heterophils) and remained decreased (P<0.05) up to 48hr. However, the loss of lysine in the cellular component of the immune system was dwarfed by the gain in lysing in acute phase proteins. Lysine accretion in lymphocyte subsets increased between 3d to 14d post-injection (P<0.05). Lysine accretion in E. coli-specific immunoglobulins (IgM and IgY) increased 5d and 7d post-injection but was quantitatively negligible. The lysine content of the systemic components of the immune system relative to all of the lysine in an adult chicken was 0.39% at maintenance, which increased to 0.80% and 0.60% for the acute phase and adaptive phases, respectively. The entire cost of the adaptive immune response (specific antibodies and lymphocytes) is easily fueled by the decay of the acute phase proteins produced during the innate response; suggesting that adaptive immunity has no net cost. Because these numerical values can be difficult to put into biological perspective, comparison to the nutrients contained in other important tissues is instructive. The systemic immune system at maintenance has the same lysine content as 16% of a medium egg, 5% of a pectoralis muscle, or 332 average sized feathers. Glycine, cysteine, serine, and threonine were used for the immune response in a greater relative proportion than lysine and are predicted to be limiting in a diet formulated for growth.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Cornax, I., R.L. Walzem, C. Larner, R.D. Macfarlane, and K.C. Klasing. 2013. Mobilization of ectopic yolk in Gallus gallus domesticus: a novel reverse lipid transport process. The Journal of experimental biology 216: 1949-1958
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Davin, R., E.G. Manzanilla, K.C. Klasing, and J.F. Perez. 2012. Evolution of zinc, iron, and copper concentrations along the gastrointestinal tract of piglets weaned with or without in-feed high doses of zinc oxide compared to unweaned littermates. Journal of animal science 90 Suppl 4: 248-250.
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Schusser, B., E.J. Collarini, H. Yi, S.M. Izquierdo, J. Fesler, D. Pedersen, K.C. Klasing, B. Kaspers, W.D. Harriman, M.C. van de Lavoir, R.J. Etches, and P.A. Leighton. 2013. Immunoglobulin knockout chickens via efficient homologous recombination in primordial germ cells. Proceedings of the National Academy of Sciences of the United States of America 110: 20170-20175.
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Iseri, V.J., and K.C. Klasing. 2013. Dynamics of the systemic components of the chicken (Gallus gallus domesticus) immune system following activation by Escherichia coli; implications for the costs of immunity. Developmental and comparative immunology 40: 248-257.
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Progress 01/01/12 to 12/31/12
Outputs
OUTPUTS: We continue our investigations on the effect of immune protection on the performance of chickens and other birds. Immune systems must protect against pathogens but the nutritional costs of developing, maintaining and using this system impinge on the flow of nutrients to growth and reproduction. Thus immune systems have been selected to be efficient. We have used a variety of chicken genetic lines, as well as other avian species to untangle the costs of immunity and the selective forces that modify these expenditures. In adult chickens, the systemic immune system (including all major leukocytes and protective proteins but excluding mucosal leukocytes) at maintenance has the same lysine content as 16% of a medium egg, 332 average sized feathers, or 5% of a pectoralis muscle. The increase in lysine accretion due to injection of E. coli was 113% during the acute phase (24 hrs) and 44% during the adaptive response (5 d). Acute phase proteins contributed 84% to the lysine used during the acute phase and Ig contributed 44% during the adaptive phase. The remainder was due to leukocyte proliferation. However, the lysine accreted in the expansion of the liver for support of the acute phase response was almost 3 times the lysine accreted for the cells and proteins of the immune system, indicating that the liver is the largest contributor to the acute phase response. The adaptive response was completely fueled by the decay of the acute phase response and did not have a net nutritional cost. To determine the major evolutionary forces that affect the investment in immunity we captured approximately 200 individuals of 40taxa in tropical and temperate areas of the Americas. There was a direct positive association between body mass and the amount of energy expended and acute phase proteins produced during the acute phase response to E. coli. Smaller species invested more in constitutive aspects of immunity and less in pathogen-induced responses. We expected to find that longer lived species invested more, but this was not the case. This research approach illustrates the power of combining natural selection and artificial selection for understanding the basic principles of biology that underlie both. PARTICIPANTS: Cooperating scientists: Martin Wikelski, Max Plank, Germany, Ramesh Salvaraj, The Ohio State Univeristy, Robert Etches, Chrystal Bioscience; Irena Tieleman, Gronigen Univ; Rose Walzem, Texas A&M; Elizabeth Koutsos, Mazuri Inc., Edgar G. Manzanilla, Universitat Autonoma de Barcelona TARGET AUDIENCES: National Institutes of Health, US Government Department of the Interior, US Government Poultry Producers in North, Central, South America, Europe, Australia, Asia PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
Understanding the nutritional cost of various types of immune system has allowed poultry and swine breeding companies to optimize immunity against pathogens while simultaneously maximizing production efficiency. This approach improves animal welfare and profitability. Additionally, the human health field has been informed by these studies and has incorporated this knowledge into approaches for improving the nutrition of children in developing countries that have high rates of infectious diseases and stunted growth.
Publications
- Roura, E., Humphrey, B., Klasing, K., Swart, M., 2011. Is the pig a good umami sensing model for humans A comparative receptor study. Flavour and Fragrance Journal 26, 282-285.
- Zylberberg, M., Lee, K.A., Klasing, K.C., Hahn, T.P., Wikelski, M., 2012. Change in Avian Pox prevalence varies by species and land use type. Integrative and Comparative Biology 52, 200-209.
- Heinze, C.R., Hawkins, M.G., Gillies, L.A., Wu, X., Walzem, R.L., German, J.B., Klasing, K.C., 2012. Effect of dietary omega-3 fatty acids on red blood cell lipid composition and plasma metabolites in the cockatiel, Nymphicus hollandicus. J Anim Sci 90, 3068-3079. Klasing, K., 2011. Nutritional diseases in poultry: Role of infectious challenges., Congress of the World Veterinary Poultry Association, Cancun, Mexico, pp. 52-57.
- Klasing, K.C., 2011. Environmentally induced stochasticity of adaptive immunity. Good, bad, or ugly Integrative and Comparative Biology 51, E70-E75.
- Ozpinar, H., Aydin, I.H., Klasing, K.C., Tekiner, I.H., 2012. Interaction of Mannan oligosaccharide with Immune System "Transport of MOS in to the Lamina Propria". Kafkas Universitesi Veteriner Fakultesi Dergisi 18, 121-128.
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Progress 01/01/11 to 12/31/11
Outputs
OUTPUTS: We continue to examine effects of nutrients on the immune system. In collaboration with past graduate students Koutsos and Selvaraj, we examined the mechanism by which the immunemodulatory nutrients lutein and n-3 PUFA affect inflammation. Both lutein and n-3 PUFA were found to blunt LPS-induced PPARα and RXRα downregulation. Dietary n-3 PUFA blunted the increase in IL-1 mRNA due to LPS in a PPAR-dependent manner. The effects of lutein appear to have an epigenetic component that allows this immunomodulation and continue after withdrawal for at least one year. This mechanism exists in both commercial broilers (Cobb) and layers (HyLine). Second, we have been examining the effect the relative costs and benefits of maternal antibody allocation to yolks for both the hen and the chick. An examination of the nutritional costs indicates that negligible resources are needed to provision the egg with IgY. Injection of purified antibodies into the yolk sac of newly hatched chicks successfully elevated the plasma titres of specific anti-KLH IgY in neonates. The enhanced level of maternal antibodies had little effect on the IGY response of chicks, although the response was low even in naive chicks. Adult immune response depended largely on the magnitude of the juvenile immune response. Furthermore, the magnitude of the adult immune IgY response to a challenge with a high protein preparation of LPS was diminished by provisioning anti-KLH antibodies. This only happened when the anti-KLH IgY was injected into the egg and not when the hen was hypervaccinated against Igy. This suggests that increasing total amount of IgY in an egg diminishes novel responses. It is possible that this inhibitory effect on novel immune responses contributes the cost of provisioning an egg with high IgY levels. We are now exploring the impacts of immune status in the embryo and early in life on development of the immune system. We are using a robust dose response of adding additional IgY to the yolk on the diversification of the B cell repertoire, levels of natural antibodies and specific Igy response of chicks. Preliminary results suggest that early challenge results in growth depression and resets the maturation trajectory to stop growth at a lower body weight and begin reproduction earlier. PARTICIPANTS: Cooperating scientists: Ramesh Salvaraj, The Ohio State Univeristy, Robert Ricklefs, University of Missouri; Robert Etches, Chrystal Bioscience; Irena Tieleman, Gronigen Univ; Rose Walzem, Texas A&M; Elizabeth Koutsos, Mazuri Inc., Edgar G. Manzanilla, Universitat Autonoma de Barcelona TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Understanding the impact of non-required nutrients on the immune system has permitted the formulation of animal diets to modulate the immune system to increase resistance to specific infectious diseases. Understanding the limitations of increasing total versus specific maternal antibody will be important in understanding the development of the immune system.
Publications
- Livingston, K.A., and K.C. Klasing. 2011. Retinyl palmitate does not have an adjuvant effect on the antibody response of chicks to keyhole limpet hemocyanin regardless of vitamin A status. Poultry Science 90: 965-970.
- Klasing, K.C. 2011. Environmentally induced stochasticity of adaptive immunity. Good, bad, or ugly Integrative and Comparative Biology 51: 70-79.
- Hermes, R.G., E.G. Manzanilla, S.M. Martin-Orue, J.F. Perez, and K.C. Klasing. 2011. Influence of dietary ingredients on in vitro inflammatory response of intestinal epithelial cells challenged by an enterotoxigenic Escherichia coli (K88). Comparative immunology, microbiology and infectious diseases. 34:511-560.
- Palacios, M.G., D.W. Winkler, K.C. Klasing, D. Hasselquist, and C.M. Vleck. 2011. Consequences of immune system aging in nature: a study of immunosenescence costs in free-living Tree Swallows. Ecology 92: 952-966.5.
- Meriwether, L.S., B.D. Humphrey, D.G. Peterson, K.C. Klasing, and E.A. Koutsos. 2010. Lutein exposure, in ovo or in the diet, reduces parameters of inflammation in the liver and spleen of laying-type chicks (Gallus gallus domesticus). Journal of Animal Physiology and Animal Nutrition 94: e115-e122.
- Addison, B., R.E. Ricklefs, and K.C. Klasing. 2010. Do maternally derived antibodies and early immune experience shape the adult immune response Functional Ecology 24: 824-829.
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Progress 01/01/10 to 12/31/10
Outputs
OUTPUTS: We are trying to distinguish epigenetic from germ line influence on the interaction of nutrients on the immune system. We have used a two generational chicken model to examine the epigenetic effects. This work has resulted in the training of graduate students, presentations by these students and by Klasing at national and international conferences on animal nutrition, immunology, and evolution. Outputs have also been published in proceedings of conferences, and in peer reviewed journals. PARTICIPANTS: Nothing significant to report during this reporting period. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Dietary lutein and PUFA fat were found to be anti-inflammatory due to modification of immune tissue lutein content, PPAR, RXR isomers and IL-1beta mRNA levels in liver and spleen. This action appears to have an epigenetic component that allows this immunomodulation to continue during an animal's lifetime. This funding resource allowed the student working on this project to buy the necessary reagents to conduct the experiments required to reach these conclusions.
Publications
- Brundige, D. R., E. A. Maga, K. C. Klasing, and J. D. Murray. 2010. Consumption of pasteurized human lysozyme transgenic goats' milk alters serum metabolite profile in young pigs. Transgenic Res 19: 563-574.
- Meriwether, L. S., B. D. Humphrey, D. G. Peterson, K. C. Klasing, and E. A. Koutsos. 2010. Lutein exposure, in ovo or in the diet, reduces parameters of inflammation in the liver and spleen laying-type chicks (gallus gallus domesticus). J Anim Physiol Anim Nutr 94: e115-122.
- Selvaraj, R. K., R. Shanmugasundaram, and K. C. Klasing. 2010. Effects of dietary lutein and PUFA on ppar and rxr isomer expression in chickens during an inflammatory response. Comp Biochem Physiol 157: 198-203.
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Progress 01/01/09 to 12/31/09
Outputs
OUTPUTS: Work continued on a two generation model of Vitamin A (VA) deficiency, which is the most common vitamin deficiency in poultry due to the ephemeral nature of retinol. With this model, hens are made marginally deficient and chicks are then fed a marginally deficient diet. Thus, a vitamin A deficiency is maintained during the entire period of embryonic development. Marginal deficient levels were chosen so that squamous metaplasia was absent in respiratory, ocular, and bursal epithelia. Activation Induced Deaminase expression in the entire bursa was decreased in the deficient chicks but their bursa had fewer lymphocytes per gram tissue. When expression of AID was examined in isolated bursacytes, AID expression was not changed by vitamin A deficiency. Interestingly, RAG1 and RAG2 were increased in bursal lymphocytes. The observation that the developing immune system is more sensitive to dietary vitamin A deficiency than squamous metaplasia will provide foundation for future work in this area. We also continued work on egg yolk peritonitis of hens, which is one of the leading causes of morbidity and mortality in laying hens and broiler breaders. We initially found that yolk is cleared from the abdominal cavity by macrophages but also found that egg yolk inhibited macrophage effector functions. In an in vivo challenge study we found that when hens were injected with E. coli and yolk, mortality was very high relative to E. coli alone. To determine if egg yolk enables E. coli pathogenesis by providing a growth-promoting medium, E. coli was incubated in vitro with egg yolk, chicken macrophages and/or chicken serum and then streaked on agar plates. Egg yolk did not increase the number of bacterial colonies and did not affect chicken macrophage antibacterial function in vitro. Egg yolk did inhibit the antimicrobial activity of chicken serum. To determine how egg yolk affects the acute phase response to E. coli, the first 24 hours of the immune response was monitored in chicks injected with egg yolk and/or E. coli. Egg yolk significantly reduced the E. coli-induced inflammatory reaction, by decreasing blood heterophil: lymphocyte ratios and inhibiting IL-1BETA and IL-6 expression in the spleen. PARTICIPANTS: The following graduate students and post docs received research training in the conduct of this research: Kim Livingston, Ingrid Edwards, Kelly Lee and Edgar Garcia. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Decreased numbers of developing lymphocytes in the bursa is a more sensitive indicator of marginal vitamin A deficiency than squamous metaplasia or immunoglobulin levels. Thus, future estimates of the vitamin A requirement should be based on bursal lymphocyte demographics and not on metaplasia. Our initial estimates of vitamin A needs based on bursal morphometrics will inform future NRC recommendations for recommended dietary levels.
Publications
- Kogut MH, Klasing K. An immunologist's perspective on nutrition, immunity, and infectious diseases. Appl Poult Res. 2009;18:103-10.
- Hernandez J, Soto-Canevett E, Pinelli-Saavedra A, Resendiz M, Moya-Camarena SY, Klasing KC. In vitro effect of vitamin E on lectin-stimulated peripheral blood mononuclear cells. Vet Immunol Immunopathol. 2009 Sep 15;131:9-16.
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Progress 01/01/08 to 12/31/08
Outputs
OUTPUTS: Work was done to determine the priority of leukocyte populations for key anabolic nutrients by examining the types and amounts of transporters expressed. Bursal cells have a high priority for glucose, branched-chain amino acids and lysine, but thymic cells have a very low priority relative to other types of leukocytes and most other tissue types. Furthermore, in the face of a deficit, bursal cells upregulate their ability to obtain glucose and amino acids, whereas thymocytes downregulate their uptake. Thus, the thymus is very sensitive to periods of food deprivation, energy deficiency, or amino acid deficiency, which cause a rapid decline in cellularity and weight. Fewer CD4+ T cells results in lower IgG production, while fewer CD8+ T cells results in diminished delayed-type hypersensitivity. Research was also initiated to identify key non-anabolic nutrients that serve as regulators of immune responses. It was found that nutrients that have strong immunomodulating activities include long chain polyunsaturated fatty acids, carotenoids, secondary plant compounds (e.g. genistein, flavonoids and many others found in herbals) and vitamins A, C, D, and E. These nutrients modulate the immune system by serving as ligands for nuclear receptors, including RXR, RXR, and PPAR. PARTICIPANTS: Kirk C. Klasing, Principle Investigator Roseline Holt, Staff Research Associate TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
For nutrients that the immune system is most vulnerable due to a low priority for acquisition, requirements set based on maximal weight gain or egg production are likely to be inadequate for optimal disease resistance. Among immunomodulatory nutrients, the dose response relationship of these nutrients is non-linear and there are many unpredictable interactions between nutrients on immunity.
Publications
- Buyse, J., Swennen, Q., Niewold, T.A., Klasing, K.C., Janssens, G.P., Baumgartner, M. and Goddeeris, B.M. (2007) Dietary l-carnitine supplementation enhances the lipopolysaccharide-induced acute phase protein response in broiler chickens. Vet Immunol Immunopathol 118, 154-9.
- Millet, S., Bennett, J., Lee, K.A., Hau, M. and Klasing, K.C. (2007) Quantifying and comparing constitutive immunity across avian species. Dev Comp Immunol 31, 188-201.
- Brundige, D.R., Maga, E.A., Klasing, K.C. and Murray, J.D. (2008) Lysozyme transgenic goats' milk influences gastrointestinal morphology in young pigs. J Nutr 138, 921-6.
- Buyse, J., Swennen, Q., Vandemaele, F., Klasing, K.C., Niewold, T.A., Baumgartner, M. and Goddeeris, B.M. (2008) Dietary beta-hydroxy-beta-methylbutyrate supplementation influences performance differently after immunization in broiler chickens. J Anim Physiol Anim Nutr (Berl).
- Blatchford, R.A., Klasing, K.C., Shivaprasad, H.L., Wakenell, P.S., Archer, G.S. and Mench, J.A. (2009) The effect of light intensity on the behavior, eye and leg health, and immune function of broiler chickens. Poult Sci 88, 20-8.
- Klasing, K.C. (2009) Minimizing amino acid catabolism decreases amino acid requirements. J Nutr 139, 11-2.
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Progress 01/01/07 to 12/31/07
Outputs
OUTPUTS: In order to avoid both starvation and disease, animals must allocate resources between nutrient reserves and immune defense. We used dynamic mechanistic modeling investigate the optimal allocation. We found that animals with low reserves choose to allocate less to defense than animals with higher reserves because when reserves are low it is more important to increase reserves to reduce the risk of starvation in the future. In general, investment in immune defense increases monotonically with nutrient reserves. An exception is when the animal can reduce its probability of death from disease by reducing its food consumption. In this case, allocation to immune defense can peak at intermediate reserves. When the nutrient quality of food changes over time, the optimal response depends on the frequency of changes. We show that the strength of selection on reserve-dependent immune defense depends on how nutrient intake and partitioning to immune defense determine the probability of
death from disease. We also examined the specific effect of the nutrient lutein supplied from maternal (i.e., in ovo) and dietary routes effect cutaneous basophil hypersensitivity responses in chickens. The PHA-induced basophilic response occurred more rapidly in chicks from carotenoid-deplete eggs, but eventually became greater in chicks from carotenoid-replete eggs. Differences in leukocyte infiltration occurred due to diet and in ovo carotenoid exposure, and indicate that pre- and post-hatch carotenoid exposure had additive or synergistic effects on the PHA-induced wing web response. Evaluation of the cellular contents of the injection site is a much better indicator of the immunomodulatory effects of lutein than measurements of the amount of swelling.
PARTICIPANTS: Kirk C. Klasing, Arash Naziripour, Laura Flatow, Kimberly Livinstone. Collaborators: Pat Wakenell
TARGET AUDIENCES: Animal producers and animal biologists.
PROJECT MODIFICATIONS: None
Impacts
Infectious diseases reduce productivity and diminish animal welfare. Appropriate nutrition may aid in minimizing the incidence of diseases by enhancing immunity. An understanding of the pressures imposed by evolution that underlie poultry nutrition as well as those which underlie immunity provides focus to the field of nutritional immunology. Additional understanding is provided by knowing the specific cellular mechanisms by which diet affects immunity, and how these mechanisms pertain to specific nutrients and pathogens. These approaches indicate that higher inclusion rates of all nutrients are not always better and that the traditional ideas of adding a surfeit of specific nutrients to improve immunity does not usually serve us well. Understanding the nuances of nutrition and immunity is important for optimizing bird health and productivity, and will be an important contributor towards fulfilling the consumer's conflicting demands for more natural production and
better animal welfare.
Publications
- Houston, A.I., McNamara, J.M., Barta, Z. and Klasing, K.C. (2007) The effect of energy reserves and food availability on optimal immune defence. Proc Biol Sci 274, 2835-42.
- Klasing, K.C. (2007) Nutrition and the immune system. Br Poult Sci 48, 525-37.
- Koutsos, E.A., Garcia Lopez, J.C. and Klasing, K.C. (2007) Maternal and dietary carotenoids interactively affect cutaneous basophil responses in growing chickens (gallus gallus domesticus). Comp Biochem Physiol B Biochem Mol Biol 147, 87-92.
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Progress 01/01/06 to 12/31/06
Outputs
We have examined the relative priority of different leukocyte populations for glucose and for basic amino acids (lysine and arginine) relative to the priority of other tissues, including the heart, skeletal muscle, liver, and brain. This was done by quantifying the expression of both high and low priority isoforms of the glucose transporter (Gluc 1-3) and the cationic amino acid transporter (CAT 1-3). By comparing the expression levels of low versus high priority transporters we were able to establish the priority of bursal, thymic, and splenic lymphocytes for energy and amino acids. Bursal cells had a very high priority for these nutrients, similar to that of heart and brain. Thymic cells had a very low priority, similar to muscle. Bursal cells increased their expression of high priority transporters during nutrient deficiency, permitting nutrients to be obtained even when circulating levels were deficient. Conversely, thymocytes decreased their expression of high
priority receptors during a deficiency, resulting in starvation and apoptosis.
Impacts
This research provides a roadmap for identifying diets that diminish the immunity and disease resistance of poultry. Because developing thymocytes do not express sufficient high priority receptors they can not compete with skeletal muscle for nutrients. During a nutrient deficiency, developing B cells augment their priority whereas developing T cells decrease theirs. Consequently, developing T cells are exceptionally vulnerable to nutrient deficiencies and can be used as sentinels for mal-nutrition.
Publications
- Blackmore, C., K. Klasing, and P. Wakenell, 2006. Effect of infectious bursal disease virus insult on iron, copper, and zinc concentration in liver, bursa of Fabricius, spleen, pancreas, and serum of chickens. Avian Dis, 50(2): p. 303-5.
- Humphrey, B.D., C.B. Stephensen, C.C. Calvert, and K.C. Klasing, 2006. Lysine deficiency and feed restriction independently alter cationic amino acid transporter expression in chickens (Gallus gallus domesticus). Comp Biochem Physiol A Mol Integr Physiol, 143(2): p. 218-27.
- Koutsos, E.A., J.C. Garcia Lopez, and K.C. Klasing, 2006. Carotenoids from in ovo or dietary sources blunt systemic indices of the inflammatory response in growing chicks (Gallus gallus domesticus). J Nutr, 136(4): p. 1027-31.
- Martin, L.B., P. Han, J. Lewittes, K.C. Klasing, and M. Wikelski, Phytohemagglutinin-induced skin swelling in birds: Histological support for a classic immunoecological technique. Functional Ecology, 2006. 20: p. 290-299.
- Selvaraj, R.K. and K.C. Klasing, 2006. Lutein and eicosapentaenoic acid interact to modify iNOS mRNA levels through the PPARgamma/RXR pathway in chickens and HD11 cell lines. J Nutr, 136(6): p. 1610-6.
- Selvaraj, R.K., E.A. Koutsos, C.C. Calvert, and K.C. Klasing, 2006. Dietary lutein and fat interact to modify macrophage properties in chicks hatched from carotenoid deplete or replete eggs. J Anim Physiol Anim Nutr (Berl), 90(1-2): p. 70-80.
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Progress 01/01/05 to 12/31/05
Outputs
Lutein, a dihydroxycarotenoid, has antioxidant and immunomodulatory potential in certain situations. Using three 2 x 2 x 2 factorial designs, carotenoid exposure during in ovo embryogenesis and via the diet post-hatch was examined for effects on the inflammatory response to lipopolysaccharide (LPS). LPS increased splenic and hepatic mRNA expression for inducible nitric oxide synthase (iNOS), interleukin-1 (IL-1), IL-6, and transforming growth factor beta2 (TGFbeta2) (P<0.01), increased plasma haptoglobin and Zn (P<0.01), and reduced plasma Fe and Cu (P<0.01). After LPS challenge, chicks from carotenoid-replete eggs had greater liver iNOS mRNA expression (P<0.05) than chicks from deplete eggs, while chicks from carotenoid-deplete eggs had greater changes in plasma minerals (P<0.05 for Zn, Fe, Mg, S and Cu). Compared to chicks fed 40 mg lutein/kg diet, chicks fed 0 mg lutein had significant reductions in body weight gains, but higher haptoglobin and relative weights of
thymus, bursa and spleen post-LPS (P<0.05). Chicks from carotenoid-deplete eggs fed 40 mg lutein had LPS-induced increases in plasma K and Na (P<0.05) but not splenic iNOS mRNA expression, and tended to have increased splenic and hepatic TGFbeta mRNA (P<0.08) expression compared to birds hatched from carotenoid-replete eggs and fed 40 mg lutein. Data suggest that a lack of carotenoid exposure, either in ovo or post-hatch, increases parameters of systemic inflammation. Follow up experiments were conducted to investigate the effect of lutein and fat on inducible nitric oxide synthase (iNOS), peroxisome proliferator activated receptors (PPAR) alpha, beta and gamma, retinoic acid X receptor (RXR) alpha and gamma mRNA levels. Macrophages were collected from broiler chickens fed 3 or 6 % dietary fatwith 0, 25 and 50mg lutein /kg feed for 23d. In the final experiment, a 3 X 3 factorial, eicosapentaenoic acid at 0, 15 and 50micro-mol/L and lutein at 0, 10 and 100micro-mol/L, were applied to
HD11 cell culture for 24 h. Lutein and fat or EPA interacted to affect iNOS (P =0.01, P <0.01), PPARgamma (P =0.05, P <0.01) and RXRalpha (P =0.03, P =0.01) mRNA levels in macrophages and HD11 cells, respectively. At 3% dietary fat or up to 15 micro-mol/L media EPA, increasing lutein increased the iNOS mRNA. However, at 6% dietary fat or 50 micro-mol/L EPA, lutein did not cause a rise in iNOS mRNA. Increasing media lutein from 0 to 100micro-mol/L decreased the iNOS mRNA. Increasing the lutein with high fat (6%) or EPA (15micro-mol/L EPA) increased PPARgamma and RXRalpha mRNA levels. Lutein increased PPARalpha mRNA levels in both macrophages (P <0.01) and HD11 (P =0.01) cells and RXRgamma (P <0.01) mRNA levels in macrophages. GW9662, a PPARgamma antagonist, in HD11 cells prevented (P <0.01) the lutein induced iNOS mRNA down regulation. LG101208, a RXR antagonist, prevented (P <0.01) 10 micro-mol/L lutein induced iNOS upregulation and 100 micro-mol/L lutein induced iNOS mRNA down
regulation. It is concluded that lutein and EPA interact through PPARgamma pathway to modulate iNOS mRNA
Impacts
Diet modifies immune responses and, consequently, susceptibility to infectious diseases. Nutrition can be used to optimize immunity to specific pathogens and to mitigate the production losses that are associated with robust immune responses to vaccines and pathogens. In practice, adoption of nutritional immunomodulation has been slow, largely because the results of simple nutritional interventions used in research do not hold up when applied to different genetic backgrounds and variable diets used in the industry. Our results show that there are multifaceted interactions between immunomodulatory nutrients that are not always predictable from the effects of individual nutrients. Dietary fatty acids, lutein, vitamins A and E are the major immunomodulatory nutrients and were shown to exert their effect through the action of nuclear hormone receptors, whose activation down regulates the NFkB/AP-1 transcription factors in macrophages and other regulatory leukocytes. We
further identified the nuclear hormone receptors that mediate the synergistic and antagonistic interactions of nutrients. This knowledge allows us to design diets that will effect predictable changes in the balance of the immune response so as to maximize disease resistance and animal productivity.
Publications
- Humphrey, B.D., Calvert, C.C. and Klasing, K.C. (2004) The ratio of full length IgY to truncated IgY in immune complexes affects macrophage phagocytosis and the acute phase response of mallard ducks (anas platyrhynchos). Dev Comp Immunol 28, 665-72.
- Humphrey, B.D., Stephensen, C.B., Calvert, C.C. and Klasing, K.C. (2004) Glucose and cationic amino acid transporter expression in growing chickens (gallus gallus domesticus). Comp Biochem Physiol A Mol Integr Physiol 138, 515-25.
- Humphrey, B.D., Stephensen, C.B., Calvert, C.C. and Klasing, K.C. (2004) Glucose and cationic amino acid transporter expression in growing chickens (gallus gallus domesticus). Comp Biochem Physiol A Mol Integr Physiol 138, 515-25.
- Klasing, K.C. (2004) The costs of immunity. Acta Zoologica Sinica 50, 961-969.
- Humphrey, B.D. and Klasing, K.C. (2005) The acute phase response alters cationic amino acid transporter expression in growing chickens (gallus gallus domesticus). Comp Biochem Physiol A Mol Integr Physiol.
- Klasing, K.C. (2005) Potential impact of nutritional strategy on noninvasive measurements of hormones. Ann N Y Acad Sci 1046, 5-16.
- Matson, K.D., Ricklefs, R.E. and Klasing, K.C. (2005) A hemolysis-hemagglutination assay for characterizing constitutive innate humoral immunity in wild and domestic birds. Dev Comp Immunol 29, 275-86.
- Mireles, A.J., Kim, S.M. and Klasing, K.C. (2005) An acute inflammatory response alters bone homeostasis, body composition, and the humoral immune response of broiler chickens. Poultry Science 84, 553-560.
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