, Volume 7, Issue 5–6, pp 449–459 | Cite as

Simultaneous evaluation of circulating chemokine and cytokine profiles in elderly subjects by multiplex technology: relationship with zinc status

  • Erminia MarianiEmail author
  • Luca Cattini
  • Simona Neri
  • Marco Malavolta
  • Eugenio Mocchegiani
  • Giovanni Ravaglia
  • Andrea Facchini
Research Article


Cellular components of both adaptive and innate immune systems produce different chemokines and cytokines, involved in different signalling pathways among cells, and modulate effector function during immune response, playing a key role in the regulation of the type and extent of the immune response in the elderly. We evaluated the circulating concentration of selected chemokines: MCP-1, MIP-1α, IL-8, RANTES together with IL-6 and TNF-α in plasma obtained from a group of healthy old subjects, in order to highlight possible differences in the synthesis of these factors, assuming that both the cytokine and the chemokine networks are remodelled with ageing. The simultaneous evaluation was performed by a multiplex analysis system. In addition, since micronutrient deficiency may underlie an inflammatory response, the association between chemokine levels and a nutritional element such as zinc was also evaluated, since the immune system is the first system to be affected by changing zinc levels, due to its high cell turnover. A progressive age-related increase of plasma concentrations of all soluble factors was observed. The increment was particularly evident for IL-6, IL-8, MCP-1 and TNF-α in the over 85-year-old group in concomitance with increasing percentages of subjects with low circulating levels of zinc. In conclusion, the remodelling of chemokine profiles, skewed to Th2 response by both advanced ages and circulating levels of zinc, might reflect different states of activation and/or responsiveness of the human immune cell/mediator network, thus influencing the ability to develop rapid innate and long-lasting adaptive immune responses with advancing age.


Chemokine profiles Inflammation Ageing Zinc Multiplex immunoassay Bio-Plex™ 



This work was partially supported by grants from Bologna University (60% fund), Ricerca Corrente IOR, Italian Health Ministry fund, and was performed under the aegis of the EU ZINCAGE project (FOOD-CT-2003-506850). The authors thank Mrs Patrizia Rappini and Graziella Salmi for typing assistance.


  1. Antonelli A, Rotondi M, Fallahi P et al (2006) Increase of CXC chemokine CXCL10 and CC chemokine CCL2 serum levels in normal ageing. Cytokine 34:32–38PubMedCrossRefGoogle Scholar
  2. Baggiolini M (1998) Chemokines and leukocyte traffic. Nature 392:565–568PubMedCrossRefGoogle Scholar
  3. Brod SA (2000) Unregulated inflammation shortens human functional longevity. Inflamm Res 49:561–570PubMedCrossRefGoogle Scholar
  4. Cazzola M, Polosa R (2006) Anti-TNFα and Th1 cytokine-directed therapies for the treatment of asthma. Curr Opin Allergy Clin Immunol 6:43–50PubMedCrossRefGoogle Scholar
  5. Deo R, Khera A, McGuire DK et al (2004) Association among plasma levels of monocyte chemoattractant protein-1, traditional cardiovascular risk factors and subclinical atherosclerosis. J Am Coll Cardiol 44:1812–1818PubMedCrossRefGoogle Scholar
  6. Facchini A, Mariani E, Mariani AR et al (1987) Increased number of circulating Leu 11+ (CD 16) large granular lymphocytes and decreased NK activity during human ageing. Clin Exp Immunol 68:340–347PubMedGoogle Scholar
  7. Fraker PJ, King LE (2004) Reprogramming of the immune system during zinc deficiency. Annu Rev Nutr 24:277–298PubMedCrossRefGoogle Scholar
  8. Gabriel P, Cakman I, Rink L (2002) Overproduction of monokines by leukocytes after stimulation with lipopolysaccharide in the elderly. Exp Gerontol 37:235–247PubMedCrossRefGoogle Scholar
  9. Gerli R, Monti D, Bistoni O et al (2000) Chemokines, sTNF-Rs and sCD30 serum levels in healthy aged people and centenarians. Mech Ageing Dev 121:37–46PubMedCrossRefGoogle Scholar
  10. Gu L, Tseng S, Horner RM et al (2000) Control of Th2 polarization by the chemokine monocyte chemoattractant protein-1. Nature 404:407–411PubMedCrossRefGoogle Scholar
  11. Hambidge M (2000) Human zinc deficient. J Nutr 130:1344S–1349SPubMedGoogle Scholar
  12. Ibs KH, Rink L (2003) Zinc-altered immune function. J Nutr 133:1452S–1456SPubMedGoogle Scholar
  13. Inadera H, Egashira K, Takemoto M et al (1999) Increase in circulating levels of monocyte chemoattractant protein-1 with aging. J Interferon Cytokine Res 19:1179–1182PubMedCrossRefGoogle Scholar
  14. Licastro F, Candore G, Lio D et al (2005) Innate immunity and inflammation in ageing: a key for understanding age-related diseases. Immun Ageing 2:1–14CrossRefGoogle Scholar
  15. Ligthart GJ, Corberand JX, Fournier C et al (1984) Admission criteria for immunogerontological studies in man: the SENIEUR protocol. Mech Ageing Dev 28:47–55PubMedCrossRefGoogle Scholar
  16. Linton PJ, Dorshkind K (2004) Age-related changes in lymphocyte development and function. Nat Immunol 5:133–139PubMedCrossRefGoogle Scholar
  17. Malavolta M, Costarelli L, Giacconi R, Muti E, Bernardini G, Tesei S, Cipriano C, Mocchegiani E (2006) Single and three-colours flow cytometry assay for intracellular zinc ion availability in human lymphocytes with Zinpyr-1 and double immunofluorescence: relationship with metallothioneins. Cytometry A (in press)Google Scholar
  18. Mariani E, Facchini A (2003) Characterisation of NK cells in the elderly. In: Pawelec G (ed) Advances in cell ageing and gerontology series, vol. 13. Basic biology and clinical impact of immunosenescence. Elsevier BV, Amsterdam, pp. 133–157Google Scholar
  19. Mariani E, Roda P, Mariani AR et al (1990) Age-associated changes in CD8+ and CD16+ cell reactivity: clonal analysis. Clin Exp Immunol 81:479–484PubMedCrossRefGoogle Scholar
  20. Matsusaka T, Fujikawa K, Nishio Y et al (1993) Transcription factors NK-IL6 and NK-kappa B synergistically activate transcription of the inflammatory cytokines, interleukin 6 and interleukin 8. Proc Natl Acad Sci USA 90:10193–10197PubMedCrossRefGoogle Scholar
  21. Maugeri D, Russo MS, Franze C et al (1998) Correlations between C-reactive protein, interleukin-6, tumor necrosis factor-alpha and body mass index during senile osteoporosis. Arch Gerontol Geriatr 27:159–163CrossRefPubMedGoogle Scholar
  22. Maurer M, von Stebut E (2004) Molecules in focus. Macrophage inflammatory protein-1. Int J Biochem Cell Biol 36:1882–1886PubMedCrossRefGoogle Scholar
  23. Menten P, Wuyts A, Van Damme J (2002) Macrophage inflammatory protein-1. Cytokine Growth Factor Rev 13:455–481PubMedCrossRefGoogle Scholar
  24. Moser B, Williman K (2004) Chemokines: role in inflammation and immune surveillance. Ann Rheum Dis 63:ii84–ii89PubMedCrossRefGoogle Scholar
  25. Niemir ZI, Stein H, Ciechanowicz A (2004) The in situ expression of interleukin-8 in the normal human kidney and in different morphological forms of glomerulonephritis. Am J Kidney Dis 43:983–998PubMedCrossRefGoogle Scholar
  26. Olivieri F, Bonafè M, Cavallone L et al (2002) The-174 C/G locus affects in vitro/in vivo IL-6 production during aging. Exp Gerontol 37:309–314PubMedCrossRefGoogle Scholar
  27. Ono JS, Nakamura T, Miyazaki D et al (2003) Chemokines: roles in leukocyte development, trafficking and effector function. J Allergy Clin Immunol 111:1185–1199PubMedCrossRefGoogle Scholar
  28. Paganelli R, Scala E, Rosso R et al (1996) A shift to Th0 cytokine production by CD4+ cells in human longevity: studies on two healthy centenarians. J Immunol 26:2030–2034Google Scholar
  29. Paganelli R, Di Iorio A, Patricelli L et al (2002) Proinflammatory cytokines in sera of elderly patients with dementia: levels in vascular injury are higher than those of mild–moderate Alzheimer’s disease patients. Exp Gerontol 37:257–263PubMedCrossRefGoogle Scholar
  30. Pawelec G, Solana R, Remarque E et al (1998) Impact of aging on innate immunity. J Leukoc Biol 64:703–712PubMedGoogle Scholar
  31. Pawelec G, Barnett Y, Forsey R et al (2002) T cells and ageing, January 2002 update. Front Biosci 7:d1056–d1183PubMedGoogle Scholar
  32. Plowden J, Renshaw-Hoelscher M, Engleman C et al (2004) Innate immunity in ageing: impact on macrophage function. Aging Cell 3:161–167PubMedCrossRefGoogle Scholar
  33. Podkowka R, Wisniewska J, Korybalska K et al (2002) Plasma level of interleukin-6 and interleukin-8 in the elderly. Przegl Lek 59:230–233PubMedGoogle Scholar
  34. Prasad AS (2000) Effects of zinc deficiency on Th1 and Th2 cytokine shifts. J Infect Dis 182:S62–S68PubMedCrossRefGoogle Scholar
  35. Pulsatelli L, Meliconi R, Mazzetti I et al (2000) Chemokine production by peripheral blood mononuclear cells in elderly subjects. Mech Ageing Dev 121:89–100PubMedCrossRefGoogle Scholar
  36. Ravaglia G, Forti P, Maioli F et al (2000) Effect of micronutrient status on natural killer cell immune function in healthy free-living subjects aged ≥90 y1–3. Am J Clin Nutr 71:590–598PubMedGoogle Scholar
  37. Rink L, Gabriel P (2000) Zinc and the immune system. Proc Nutr Soc 59:541–552PubMedCrossRefGoogle Scholar
  38. Sanjiv AL, Cyster JG (2001) Chemokines as regulators of T cell differentiation. Nat Immunol 2:102–107CrossRefGoogle Scholar
  39. Sarkar D, Fisher PB (2006) Molecular mechanisms of aging-associated inflammation. Cancer Lett 236:13–23PubMedCrossRefGoogle Scholar
  40. Shankar AH, Prasad AS (1998) Zinc and immune function: the biological basis of altered resistance to infection. Am J Clin Nutr 68:447S–463SPubMedGoogle Scholar
  41. Solana R, Mariani E (2000) NK and NK/T cells in human senescence. Vaccine 18:1613–1620PubMedCrossRefGoogle Scholar
  42. Stellwagen D, Malenka RC (2006) Synaptic scaling mediated by glial TNF-alpha. Nature 440:1054–1059PubMedCrossRefGoogle Scholar
  43. Tieri P, Valensin S, Latora V et al (2005) Quantifying the relevance of different mediators in the human immune cell network. Bioinformatics 21:1639–1643PubMedCrossRefGoogle Scholar
  44. Wieczorowska-Tobis K, Niemir ZI, Podkowka R et al (2006) Can an increased level of circulating IL-8 be a predictor of human longevity? Med Sci Monit 12:CR118–CR121PubMedGoogle Scholar
  45. Wong MM, Fish EN (2003) Chemokines: attractive mediators of the immune response. Semin Immunol 15:5–14PubMedCrossRefGoogle Scholar
  46. World Health Organization (1996) Zinc. In: trace elements in human nutrition and health. WHO, Geneve, pp. 72-104Google Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Erminia Mariani
    • 1
    • 2
    Email author
  • Luca Cattini
    • 1
  • Simona Neri
    • 1
  • Marco Malavolta
    • 3
  • Eugenio Mocchegiani
    • 3
  • Giovanni Ravaglia
    • 4
  • Andrea Facchini
    • 1
    • 2
  1. 1.Laboratorio di Immunologia e GeneticaIstituto di Ricerca Codivilla-Putti, IOR, Via di Barbiano 1/10BolognaItaly
  2. 2.Dipartimento di Medicina Interna e GastroenterologiaUniversity of BolognaBolognaItaly
  3. 3.Immunology Center (Nutrition, Immunity and Ageing Section)INRCAAnconaItaly
  4. 4.Dipartimento di Medicina Interna Cardioangiologia ed EpatologiaUniversity of BolognaBolognaItaly

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