European Journal of Nutrition

, Volume 56, Issue 2, pp 557–567 | Cite as

Zinc enhances the number of regulatory T cells in allergen-stimulated cells from atopic subjects

  • Eva Rosenkranz
  • Ralf-Dieter Hilgers
  • Peter Uciechowski
  • Arnd Petersen
  • Birgit Plümäkers
  • Lothar Rink
Original Contribution

Abstract

Purpose

The trace element zinc is essential for immune function and its regulation. Since zinc deficiency and allergic hyperresponsive reactions are often accompanied, the influence of zinc on allergen-induced cell growth, CD4+ regulatory T (Treg) cell numbers and cytokine expression during allergic immune reactions was investigated.

Methods

Peripheral blood mononuclear cells (PBMCs) from non-atopic and atopic subjects were treated with timothy grass allergen pre-incubated with or without zinc. Proliferation was determined by analyzing the incorporation of 3H-thymidine. Intracellular zinc and Foxp3 levels and cell surface antigens were measured by FACS, cytokine expression by ELISA and real-time PCR.

Results

Incubation with 50 μM zinc sulfate (Zn50) enhances cytosolic zinc concentrations in CD3+ T cells. The data also reveal that the combination of Zn50 plus allergen significantly reduces PBMC proliferation of atopic subjects. Additionally, Zn50 plus allergen enhances Th1 cytokine responses shown by increased interferon (IFN)-γ/interleukin (IL)-10 ratios as well as enhanced tumor necrosis factor-α release. In response to allergen, zinc increases Treg cells and upregulates the mRNA expression of cytotoxic T-lymphocyte antigen-4 in atopic subjects. Interestingly, Zn50 alone leads to an increase of CD4+CD25high(hi)+ cells in atopic and non-atopic subjects.

Conclusions

Zinc may regulate unwanted hyperresponsive immune reactions by suppressing proliferation through a significant shift from IL-10 to the Th1 cytokine IFN-γ, and enhanced regulatory T cell numbers. Therefore, zinc supplementation may be a promising tool for the therapy of allergies, without negatively affecting the immune system.

Keywords

Zinc Nutritional immunology Allergy Regulatory T cells Treg Foxp3 

Notes

Acknowledgments

We acknowledge grant support SFB/TR22 A03 and Z01 from the Deutsche Forschungsgemeinschaft for A.P. L.R. is a member of the European COST action Zinc-Net (TDI304).

Compliance with ethical standards

The study was approved by the local ethical committee (No. AZ 016/09) and had therefore been performed in accordance with the ethical standards laid down in the 1964 Declaration of Helsinki and its later amendments. All persons gave their informed consent prior to their inclusion in the study.

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Maywald M, Rink L (2015) Zinc homeostasis and immunosenescence. J Trace Elem Med Biol 29:24–30CrossRefGoogle Scholar
  2. 2.
    Haase H, Rink L (2014) Zinc signals and immune function. BioFactors 40:27–40CrossRefGoogle Scholar
  3. 3.
    Andreini C, Banci L, Bertini I, Rosato A (2006) Counting the zinc-proteins encoded in the human genome. J Proteome Res 5:196–201CrossRefGoogle Scholar
  4. 4.
    Ibs KH, Rink L (2003) Zinc-altered immune function. J Nutr 133:1452S–1456SGoogle Scholar
  5. 5.
    Prasad AS (2009) Impact of the discovery of human zinc deficiency on health. J Am Coll Nutr 28:257–265CrossRefGoogle Scholar
  6. 6.
    Rink L (2011) Zinc in human health. IOS Press, AmsterdamGoogle Scholar
  7. 7.
    Haase H, Rink L (2009) Functional significance of zinc-related signaling pathways in immune cells. Annu Rev Nutr 29:133–152CrossRefGoogle Scholar
  8. 8.
    Haase H, Rink L (2014) Multiple impacts of zinc on immune function. Metallomics 6:1175–1180CrossRefGoogle Scholar
  9. 9.
    Kahmann L, Uciechowski P, Warmuth S, Pliimdkers B, Gressner AM, Malavolta M, Mocchegiani E, Rink L (2008) Zinc supplementation in the elderly reduces spontaneous inflammatory cytokine release and restores T cell functions. Rejuvenation Res 11:227–237CrossRefGoogle Scholar
  10. 10.
    Kaltenberg J, Plum LM, Ober-Blobaum JL, Honscheid A, Rink L, Haase H (2010) Zinc signals promote IL-2-dependent proliferation of T cells. Eur J Immunol 40:1496–1503CrossRefGoogle Scholar
  11. 11.
    Honscheid A, Dubben S, Rink L, Haase H (2012) Zinc differentially regulates mitogen-activated protein kinases in human T cells. J Nutr Biochem 23:18–26CrossRefGoogle Scholar
  12. 12.
    Yu MC, Lee WW, Tomar D, Pryshchep S, Czesnikiewicz-Guzik M, Lamar DL, Li GJ, Singh K, Tian L, Weyand CM, Goronzy JJ (2011) Regulation of T cell receptor signaling by activation-induced zinc influx. J Exp Med 208:775–785CrossRefGoogle Scholar
  13. 13.
    Bhatnagar S, Wadhwa N, Aneja S, Lodha R, Kabra SK, Natchu UCM, Sommerfelt H, Dutta AK, Chandra J, Rath B, Sharma M, Sharma VK, Kumari M, Strand TA (2012) Zinc as adjunct treatment in infants aged between 7 and 120 days with probable serious bacterial infection: a randomised, double-blind, placebo-controlled trial. Lancet 379:2072–2078CrossRefGoogle Scholar
  14. 14.
    Prasad AS (2009) Zinc: role in immunity, oxidative stress and chronic inflammation. Curr Opin Clin Nutr Metab Care 12:646–652CrossRefGoogle Scholar
  15. 15.
    Kitabayashi C, Fukada T, Kanamoto M, Ohashi W, Hojyo S, Atsumi T, Ueda N, Azuma I, Hirota H, Murakami M, Hirano T (2010) Zinc suppresses T(h)17 development via inhibition of STAT3 activation. Int Immunol 22:375–386CrossRefGoogle Scholar
  16. 16.
    Kown MH, van der Steenhoven TJ, Jahncke CL, Mari C, Lijkwan MA, Koransky ML, Blankenberg FG, Strauss HW, Robbins RC (2002) Zinc chloride-mediated reduction of apoptosis as an adjunct immunosuppressive modality in cardiac transplantation. J Heart Lung Transpl 21:360–365CrossRefGoogle Scholar
  17. 17.
    Stoye D, Schubert C, Goihl A, Guttek K, Reinhold A, Brocke S, Grungreiff K, Reinhold D (2012) Zinc aspartate suppresses T cell activation in vitro and relapsing experimental autoimmune encephalomyelitis in SJL/J mice. Biometals 25:529–539CrossRefGoogle Scholar
  18. 18.
    Campo CA, Wellinghausen N, Faber C, Fischer A, Rink L (2001) Zinc inhibits the mixed lymphocyte culture. Biol Trace Elem Res 79:15–22CrossRefGoogle Scholar
  19. 19.
    Faber C, Gabriel P, Ibs KH, Rink L (2004) Zinc in pharmacological doses suppresses allogeneic reaction without affecting the antigenic response. Bone Marrow Transpl 33:1241–1246CrossRefGoogle Scholar
  20. 20.
    Shevach EM (2009) Mechanisms of Foxp3(+) T regulatory cell-mediated suppression. Immunity 30:636–645CrossRefGoogle Scholar
  21. 21.
    Sakaguchi S, Wing K, Yamaguchi T (2009) Dynamics of peripheral tolerance and immune regulation mediated by Treg. Eur J Immunol 39:2331–2336CrossRefGoogle Scholar
  22. 22.
    Takahashi T, Tagami T, Yamazaki S, Uede T, Shimizu J, Sakaguchi N, Mak TW, Sakaguchi S (2000) Immunologic self-tolerance maintained by CD25(+)CD4(+) regulatory T cells constitutively expressing cytotoxic T lymphocyte-associated antigen 4. J Exp Med 192:303–309CrossRefGoogle Scholar
  23. 23.
    Akdis CA, Akdis M (2009) Mechanisms and treatment of allergic disease in the big picture of regulatory T cells. J Allergy Clin Immunol 123:735–746CrossRefGoogle Scholar
  24. 24.
    Akdis M, Verhagen J, Taylor A, Karamloo F, Karagiannidis C, Crameri R, Thunberg S, Deniz G, Valenta R, Fiebig H, Kegel C, Disch R, Schmidt-Weber CB, Blaser K, Akdis CA (2004) Immune responses in healthy and allergic individuals are characterized by a fine balance between allergen-specific T regulatory 1 and T helper 2 cells. J Exp Med 199:1567–1575CrossRefGoogle Scholar
  25. 25.
    Antiga E, Kretz CC, Klembt R, Massi D, Ruland V, Stumpf C, Baroni G, Hartmann M, Hartschuh W, Volpi W, Del Bianco E, Enk A, Fabbri P, Krammer PH, Caproni M, Kuhn A (2010) Characterization of regulatory T cells in patients with dermatomyositis. J Autoimmun 35:342–350CrossRefGoogle Scholar
  26. 26.
    Boudousquie C, Pellaton C, Barbier N, Spertini F (2009) CD4(+)CD25(+) T cell depletion impairs tolerance induction in a murine model of asthma. Clin Exp Allergy 39:1415–1426CrossRefGoogle Scholar
  27. 27.
    Smith M, Tourigny MR, Noakes P, Catherine A, Tulic MK, Prescott SL (2008) Children with egg allergy have evidence of reduced neonatal CD4(+)CD25(+)CD127lo/− regulatory T cell function. J Allergy Clin Immunol 121:1460–1466CrossRefGoogle Scholar
  28. 28.
    Burbach GJ, Heinzerling LM, Rohnelt C, Bergmann KC, Behrendt H, Zuberbier T (2009) Ragweed sensitization in Europe—GA(2)LEN study suggests increasing prevalence. Allergy 64:664–665CrossRefGoogle Scholar
  29. 29.
    D’Amato G, Cecchi L, Bonini S, Nunes C, Annesi-Maesano I, Behrendt H, Liccardi G, Popov T, Van Cauwenberge P (2007) Allergenic pollen and pollen allergy in Europe. Allergy 62:976–990CrossRefGoogle Scholar
  30. 30.
    Provoost S, Maes T, van Durme YM, Gevaert P, Bachert C, Schmidt-Weber CB, Brusselle GG, Joos GF, Tournoy KG (2009) Decreased FOXP3 protein expression in patients with asthma. Allergy 64:1539–1546CrossRefGoogle Scholar
  31. 31.
    Ahangarani RR, Janssens W, VanderElst L, Carlier V, VandenDriessche T, Chuah M, Weynand B, Vanoirbeek JAJ, Jacquemin M, Saint-Remy JM (2009) In vivo induction of type 1-like regulatory T cells using genetically modified B cells confers long-term IL-10-dependent antigen-specific unresponsiveness. J Immunol 183:8232–8243CrossRefGoogle Scholar
  32. 32.
    de Lafaille MAC, Kutchukhidze N, Shen S, Ding Y, Yee H, Lafaille JJ (2008) Adaptive Foxp3(+) regulatory T cell-dependent and -independent control of allergic inflammation. Immunity 29:114–126CrossRefGoogle Scholar
  33. 33.
    Richter M, Bonneau R, Girard MA, Beaulieu C, Larivee P (2003) Zinc status modulates bronchopulmonary eosinophil infiltration in a murine model of allergic inflammation. Chest 123:446SCrossRefGoogle Scholar
  34. 34.
    Overbeck S, Uciechowski P, Ackland ML, Ford D, Rink L (2008) Intracellular zinc homeostasis in leukocyte subsets is regulated by different expression of zinc exporters ZnT-1 to ZnT-9. J Leukoc Biol 83:368–380CrossRefGoogle Scholar
  35. 35.
    Faneyte IF, Kristel PMP, van de Vijver MJ (2001) Determining MDR1/P-glycoprotein expression in breast cancer. Int J Cancer 93:114–122CrossRefGoogle Scholar
  36. 36.
    Haase H, Hebel S, Engelhardt G, Rink L (2006) Flow cytometric measurement of labile zinc in peripheral blood mononuclear cells. Anal Biochem 352:222–230CrossRefGoogle Scholar
  37. 37.
    Ditoro R, Capotorti MG, Gialanella G, Delgiudice MM, Moro R, Perrone L (1987) Zinc and copper status of allergic children. Acta Paediatr Scand 76:612–617CrossRefGoogle Scholar
  38. 38.
    Pawankar R, Canonica GW, Holgate ST, Lockey RF (2012) Allergic diseases and asthma: a major global health concern. Curr Opin Allergy Clin Immunol 12:39–41CrossRefGoogle Scholar
  39. 39.
    Barczyk A, Pierzchala W, Caramori G, Wiaderkiewicz R, Kaminski M, Barnes PJ, Adcock IM (2014) Decreased percentage of CD4(+)Foxp3(+)TGF-beta(+) and increased percentage of CD4(+)IL-17(+) cells in bronchoalveolar lavage of asthmatics. J Inflamm Lond 11:22CrossRefGoogle Scholar
  40. 40.
    Cohn L, Homer R, Niu N, Bottomly K (1999) Th1 cells inhibit TH2-induced airway eosinophilia and mucus production. Am J Respir Crit Care Med 159:A336Google Scholar
  41. 41.
    Hayashi N, Yoshimoto T, Izuhara K, Matsui K, Tanaka T, Nakanishi K (2007) T helper 1 cells stimulated with ovalbumin and IL-18 induce airway hyperresponsiveness and lung fibrosis by IFN-gamma and IL-13 production. Proc Natl Acad Sci USA 104:14765–14770CrossRefGoogle Scholar
  42. 42.
    Ling EM, Smith T, Nguyen XD, Pridgeon C, Dallman M, Arbery J, Carr VA, Robinson DS (2004) Relation of CD4+ CD25+ regulatory T-cell suppression of allergen-driven T-cell activation to atopic status and expression of allergic disease. Lancet 363:608–615CrossRefGoogle Scholar
  43. 43.
    Boscolo P, Di Gioacchino M, Sabbioni E, Benvenuti F, Conti P, Reale M, Bavazzano P, Giuliano G (1999) Expression of lymphocyte subpopulations, cytokine serum levels, and blood and urinary trace elements in asymptomatic atopic men exposed to an urban environment. Int Arch Occup Environ Health 72:26–32CrossRefGoogle Scholar
  44. 44.
    Guo CH, Liu PJ, Hsia S, Chuang CJ, Chen PC (2011) Role of certain trace minerals in oxidative stress, inflammation, CD4/CD8 lymphocyte ratios and lung function in asthmatic patients. Ann Clin Biochem 48:344–351CrossRefGoogle Scholar
  45. 45.
    Tudor R, Zalewski PD, Ratnaike RN (2005) Zinc in health and chronic disease. J Nutr Health Aging 9:45–51Google Scholar
  46. 46.
    Uysalol M, Uysalol EP, Yilmaz Y, Parlakgul G, Ozden TA, Ertem HV, Omer B, Uzel N (2014) Serum level of vitamin D and trace elements in children with recurrent wheezing: a cross-sectional study. BMC Pediatr 14:270CrossRefGoogle Scholar
  47. 47.
    Vural H, Uzun K, Uz E, Kocyigit A, Cigli A, Akyol O (2000) Concentrations of copper, zinc and various elements in serum of patients with bronchial asthma. J Trace Elem Med Biol 14:88–91CrossRefGoogle Scholar
  48. 48.
    Xu TF, Wang XL, Yang JZ, Hu XY, Wu WF, Guo L, Kang LD, Zhang LY (2009) Overexpression of Zip-2 mRNA in the leukocytes of asthmatic infants. Pediatr Pulmonol 44:763–767CrossRefGoogle Scholar
  49. 49.
    Picado C, Deulofeu R, Lleonart R, Agusti M, Mullol J, Qunito L, Torra M (2001) Dietary micronutrients/antioxidants and their relationship with bronchial asthma severity. Allergy 56:43–49CrossRefGoogle Scholar
  50. 50.
    Urushidate S, Matsuzaka M, Okubo N, Iwasaki H, Hasebe T, Tsuya R, Iwane K, Inoue R, Yamai K, Danjo K, Takahashi I, Umeda T, Ando S, Itai K, Nakaji S (2010) Association between concentration of trace elements in serum and bronchial asthma among Japanese general population. J Trace Elem Med Biol 24:236–242CrossRefGoogle Scholar
  51. 51.
    Ellulmicallee R, Fenech FF, Galdes A (1976) Serum zinc levels in corticosteroid-treated asthmatic-patients. Postgrad Med J 52:148–150CrossRefGoogle Scholar
  52. 52.
    Flynn A, Pories WJ, Strain WH, Hill OA, Fratianne RB (1971) Rapid serum-zinc depletion associated with corticosteroid therapy. Lancet 2:1169–1172CrossRefGoogle Scholar
  53. 53.
    Mocchegiani E, Romeo J, Malavolta M, Costarelli L, Giacconi R, Diaz LE, Marcos A (2013) Zinc: dietary intake and impact of supplementation on immune function in elderly. Age 35:839–860CrossRefGoogle Scholar
  54. 54.
    Prasad AS (2000) Effects of zinc deficiency on Th1 and Th2 cytokine shifts. J Infect Dis 182:S62–S68CrossRefGoogle Scholar
  55. 55.
    Lu HY, Xin Y, Tang Y, Shao GB (2012) Zinc suppressed the airway inflammation in asthmatic rats: effects of zinc on generation of eotaxin, MCP-1, IL-8, IL-4, and IFN-gamma. Biol Trace Elem Res 150:314–321CrossRefGoogle Scholar
  56. 56.
    Aydemir TB, Liuzzi JP, McClellan S, Cousins RJ (2009) Zinc transporter ZIP8 (SLC39A8) and zinc influence IFN-gamma expression in activated human T cells. J Leukoc Biol 86:337–348CrossRefGoogle Scholar
  57. 57.
    Cosmi L, Liotta F, Angeli R, Mazzinghi B, Santarlasci V, Manetti R, Lasagni L, Vanini V, Romagnani P, Maggi E, Annunziato F, Romagnani S (2004) Th2 cells are less susceptible than Th1 cells to the suppressive activity of CD25(+) regulatory thymocytes because of their responsiveness to different cytokines. Blood 103:3117–3121CrossRefGoogle Scholar
  58. 58.
    Walker MR, Kasprowicz DJ, Gersuk VH, Benard A, Van Landeghen M, Buckner JH, Ziegler SF (2003) Induction of FoxP3 and acquisition of T regulatory activity by stimulated human CD4(+)CD25(−) T cells. J Clin Invest 112:1437–1443CrossRefGoogle Scholar
  59. 59.
    Chen L, Feng Y, Zhou Y, Zhu W, Shen X, Chen K, Jiang H, Liu D (2010) Dual role of Zn2+ in maintaining structural integrity and suppressing deacetylase activity of SIRT1. J Inorg Biochem 104:180–185CrossRefGoogle Scholar
  60. 60.
    Plum LM, Brieger A, Engelhardt G, Hebel S, Nessel A, Arlt M, Kaltenberg J, Schwaneberg U, Huber M, Rink L, Haase H (2014) PTEN-inhibition by zinc ions augments interleukin-2-mediated Akt phosphorylation. Metallomics 6:1277–1287CrossRefGoogle Scholar
  61. 61.
    Kwon HS, Lim HW, Wu J, Schnolzer M, Verdin E, Ott M (2012) Three novel acetylation sites in the Foxp3 transcription factor regulate the suppressive activity of regulatory T cells. J Immunol 188:2712–2721CrossRefGoogle Scholar
  62. 62.
    Barron L, Dooms H, Hoyer KK, Kuswanto W, Hofmann J, O’Gorman WE, Abbas AK (2010) Cutting edge: mechanisms of IL-2-dependent maintenance of functional regulatory T cells. J Immunol 185:6426–6430CrossRefGoogle Scholar
  63. 63.
    Pan F, Yu H, Dang EV, Barbi J, Pan X, Grosso JF, Jinasena D, Sharma SM, McCadden EM, Getnet D, Drake CG, Liu JO, Ostrowski MC, Pardoll DM (2009) Eos mediates Foxp3-dependent gene silencing in CD4+ regulatory T cells. Science 325:1142–1146CrossRefGoogle Scholar
  64. 64.
    Song X, Li B, Xiao Y, Chen C, Wang Q, Liu Y, Berezov A, Xu C, Gao Y, Li Z, Wu SL, Cai Z, Zhang H, Karger BL, Hancock WW, Wells AD, Zhou Z, Greene MI (2012) Structural and biological features of FOXP3 dimerization relevant to regulatory T cell function. Cell Rep 1:665–675CrossRefGoogle Scholar
  65. 65.
    Twu YC, Teh HS (2014) The ThPOK transcription factor differentially affects the development and function of self-specific CD8(+) T cells and regulatory CD4(+) T cells. Immunology 141:431–445CrossRefGoogle Scholar
  66. 66.
    Sumitomo S, Fujio K, Okamura T, Morita K, Ishigaki K, Suzukawa K, Kanaya K, Kondo K, Yamasoba T, Furukawa A, Kitahara N, Shoda H, Shibuya M, Okamoto A, Yamamoto K (2013) Transcription factor early growth response 3 is associated with the TGF-β1 expression and the regulatory activity of CD4-positive T cells in vivo. J Immunol 191:2351–2359CrossRefGoogle Scholar
  67. 67.
    Sun X, Zhou X, Du L, Liu W, Liu Y, Hudson LG, Liu KJ (2014) Arsenite binding-induced zinc loss from PARP-1 is equivalent to zinc deficiency in reducing PARP-1 activity, leading to inhibition of DNA repair. Toxicol Appl Pharmacol 274:313–318CrossRefGoogle Scholar
  68. 68.
    Nurmatov U, Nwaru BI, Devereux G, Sheikh A (2012) Confounding and effect modification in studies of diet and childhood asthma and allergies. Allergy 67:1041–1059CrossRefGoogle Scholar
  69. 69.
    Patelarou E, Giourgouli G, Lykeridou A, Vrioni E, Fotos N, Siamaga E, Vivilaki V, Brokalaki H (2011) Association between biomarker-quantified antioxidant status during pregnancy and infancy and allergic disease during early childhood: a systematic review. Nutr Rev 69:627–641CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Eva Rosenkranz
    • 1
  • Ralf-Dieter Hilgers
    • 2
  • Peter Uciechowski
    • 1
  • Arnd Petersen
    • 3
  • Birgit Plümäkers
    • 1
  • Lothar Rink
    • 1
  1. 1.Institute of Immunology, Faculty of MedicineUniversity Hospital, RWTH Aachen UniversityAachenGermany
  2. 2.Institute of Medical Statistics, Faculty of MedicineRWTH Aachen UniversityAachenGermany
  3. 3.Division of Clinical and Molecular Allergology, Research Center BorstelAirway Research Center North (ARCN)BorstelGermany

Personalised recommendations