Journal of Clinical Immunology

, Volume 22, Issue 4, pp 195–205 | Cite as

Age-Related Decrease in the Inducibility of Heat-Shock Protein 70 in Human Peripheral Blood Mononuclear Cells

  • R. Njemini
  • M. Vanden Abeele
  • C. Demanet
  • M. Lambert
  • S. Vandebosch
  • T. MetsEmail author


We have investigated the effect of age and of the presence of proinflammatory cytokines on Hsp 70 production in human peripheral blood mononuclear cells, using flow cytometry. Twenty-seven women and 23 men, all apparently healthy, participated in the study. At 37°C, the percentage of Hsp 70-producing monocytes and lymphocytes, as well as the level of Hsp 70 in monocytes, were negatively influenced by age. After exposure of the cells to 42°C, the increase of Hsp 70 production was more pronounced in monocytes than in lymphocytes; both the intensity of Hsp 70 production and the percentage of Hsp 70-producing cells were negatively influenced by the age of the subjects, as well for monocytes as for lymphocytes. There was a negative correlation between the intensity of Hsp 70 production by monocytes exposed to 42°C and the serum levels of tumor necrosis factor-α and interleukin-6. In conclusion, in human monocytes and lymphocytes, heat-induced Hsp 70 production is reduced with increasing age and is negatively influenced in monocytes by proinflammatory cytokines.

Peripheral mononuclear blood cells heat shock heat-shock protein 70 aging proinflammatory cytokines 


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  1. 1.
    Lindquist S, Craige EA: The heat-shock proteins. Annu Rev Genet 22:631–677, 1988Google Scholar
  2. 2.
    Polla BS, Healy AM, Wojno WC, Krane SM: Hormone 1 alpha, 25-dihydroxyvitamin D3 modulates heat shock response in monocytes. Am J Physiol 252:C640–C649, 1987Google Scholar
  3. 3.
    Wu B, Hunt C, Morimoto R: Structure and expression of the human gene encoding major heat shock protein HSP70. Mol Cell Biol 5:330–341, 1985Google Scholar
  4. 4.
    Kingston RE, Schuetz TJ, Larin Z: Heat-inducible human factor that binds to a human hsp70 promoter. Mol Cell Biol 7:1530–1534, 1987Google Scholar
  5. 5.
    Moseley PL, Wallen ES, McCafferty JD, Flanagan S, Kern JA: Heat stress regulates the human 70-kDa heat-shock gene through the 3′-untranslated region. Am J Physiol 264:L533–L537, 1993Google Scholar
  6. 6.
    Gapen CJ, Moseley PL: Acidosis alters hyperthermic cytotoxicity and the cellular stress response. Therm Biol 20:321–325, 1995Google Scholar
  7. 7.
    Soti C, Csermely P: Molecular chaperones and the aging process. Biogerontol 1:225–233, 2000Google Scholar
  8. 8.
    Morimoto RI, Tissieres A, Georgopoulos C: Progress and perspectives on the biology of heat shock proteins and molecular chaperones. In The Biology of Heat Shock Proteins and Molecular Chaperones, RI Morimoto, A Tissieres, C Georgopoulos (eds). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1994 pp 1–30Google Scholar
  9. 9.
    Moore SA, Lopez A, Richardson A, Pahlavani MA: Effect of age and dietary restriction on expression of heat shock protein 70 in rat alveolar macrophages. Mech Aging Dev 104:59–73, 1998Google Scholar
  10. 10.
    Ashburner M, Bonner JJ: The induction of gene activity in drosophilia by heat shock. Cell 17:241–254, 1979Google Scholar
  11. 11.
    Burdon RH: Heat shock and the heat shock proteins. Biochem J 240:313–324, 1986Google Scholar
  12. 12.
    Lindquist S: The heat-shock response. Annu Rev Biochem 55: 1151–1191, 1986Google Scholar
  13. 13.
    Pelham HR: Speculations on the functions of the major heat shock and glucose-regulated proteins. Cell 46:959–961, 1986Google Scholar
  14. 14.
    Schlesinger MJ: Heat shock proteins: the search for functions. J Cell Biol 103:321–325, 1986Google Scholar
  15. 15.
    Dworniczak B, Mirault ME: Structure and expression of a human gene coding for a 71 kd heat shock “cognate” protein. Nucleic Acids Res 15:5181–5197, 1987Google Scholar
  16. 16.
    Ingolia TD, Craig EA: Drosophila gene related to the major heat shock-induced gene is transcribed at normal temperatures and not induced by heat shock. Proc Natl Acad Sci U S A 79:525–529, 1982Google Scholar
  17. 17.
    Lowe DG, Moran LA: Proteins related to the mouse L-cell major heat shock protein are synthesized in the absence of heat shock gene expression. Proc Natl Acad Sci USA 81:2317–2321, 1984Google Scholar
  18. 18.
    O'Malley K, Mauron A, Barchas JD, Kedes L: Constitutively expressed rat mRNA encoding a 70-kilodalton heat-shock-like protein. Mol Cell Biol 5:3476–3483, 1985Google Scholar
  19. 19.
    Sorger PK, Pelham HR: Cloning and expression of a gene encoding hsc73, the major hsp70-like protein in unstressed rat cells. EMBO J 6:993–998, 1987Google Scholar
  20. 20.
    Jaattela M, Wissing D: Heat shock proteins protect cells from monocyte cytotoxicity: Possible mechanism of self protection. J Exp Med 177:231–236, 1993Google Scholar
  21. 21.
    Chirico WJ, Waters MG, Blobel G: 70K heat shock related proteins stimulate protein translocation into microsomes. Nature 332:805–810, 1988Google Scholar
  22. 22.
    Moseley PL: Heat shock proteins and the inflammatory response. Ann NY Acad Sci 856:206–213, 1998Google Scholar
  23. 23.
    De Maio A: Heat shock proteins: facts, thoughts, and dreams. Shock 11:1–12, 1999Google Scholar
  24. 24.
    Sanchez ER, Housley PR, Pratt WB: The molybdate-stabilized glucocorticoid binding complex of L-cells contains a 98-100 kdalton steroid binding phosphoprotein and a 90 kdalton nonsteroid-binding phosphoprotein that is part of the murine heatshock complex. J Steroid Biochem 24:9–18, 1986Google Scholar
  25. 25.
    Welch WJ: Heat shock proteins functioning as molecular chaperones: Their roles in normal and stressed cells. Philos Trans R Soc Lond B Biol Sci 339:327–333, 1993Google Scholar
  26. 26.
    Bienz, M: Transient and developmental activation of heat shock genes. Trends Biochem Sci 10:157–161, 1985Google Scholar
  27. 27.
    Riabowol KT, Mizzen LA, Welch WJ: Heat shock is lethal to fibroblasts microinjected with antibodies against hsp70. Science 242:433–436, 1988Google Scholar
  28. 28.
    Johnston RN, Kucey BL: Competitive inhibition of hsp70 gene expression causes thermosensitivity. Science 242:1551–1554, 1988Google Scholar
  29. 29.
    Angelidis CE, Lazaridis I, Pagoulatos GN: Constitutive expression of heat-shock protein 70 in mammalian cells confers thermoresistance. Eur J Biochem 199:35–39, 1991Google Scholar
  30. 30.
    Li GC, Li LG, Liu YK, Mak JY, Chen LL, Lee WM: Thermal response of rat fibroblasts stably transfected with the human 70-kDa heat shock protein-encoding gene. Proc Natl Acad Sci USA 88:1681–1685, 1991Google Scholar
  31. 31.
    Fincato G, Polentarutti N, Sica A, Mantovani A, Colotta F: Expression of a heat-inducible gene of the HSP70 family in human myelomonocytic cells: regulation by bacterial products and cytokines. Blood 77:579–586, 1991Google Scholar
  32. 32.
    Polla BS, Perin M, Pizurki L: Regulation and functions of stress proteins in allergy and inflammation. Clin Exp Allergy 23:548–556, 1993Google Scholar
  33. 33.
    Yoo CG, Lee S, Lee CT, Kim YW, Han SK, Shim YS: Antiinflammatory effect of heat shock protein induction is related to stabilization of I kappa B alpha through preventing I kappa B kinase activation in respiratory epithelial cells. J Immunol 164: 5416–5423, 2000Google Scholar
  34. 34.
    LoCicero J 3rd, Xu X, Zhang L: Heat shock protein suppresses the senescent lung cytokine response to acute endotoxemia. Ann Thorac Surg 68:1150–1153, 1999Google Scholar
  35. 35.
    Hall TJ: Role of hsp70 in cytokine production. Experientia 50:1048–1053, 1994Google Scholar
  36. 36.
    Jaattela M: Overexpression of major heat shock protein hsp70 inhibits tumor necrosis factor-induced activation of phospholipase A2. J Immunol 151:4286–4294, 1993Google Scholar
  37. 37.
    Klostergaard J, Barta M, Tomasovic SP: Hyperthermic modulation of respiratory inhibition factor-and iron releasing factor-dependent macrophage murine tumor cytotoxicity. Cancer Res 49:6252–6257, 1989Google Scholar
  38. 38.
    Rao DV, Watson K, Jones GL: Age-related attenuation in the expression of the major heat shock proteins in human peripheral lymphocytes. Mech Aging Dev 107:105–118, 1999Google Scholar
  39. 39.
    Blake MJ, Udelsman R, Feulner GJ, Norton DD, Holbrook NJ: Stress-induced heat shock protein 70 expression in adrenal cortex: An adrenocorticotropic hormone-sensitive, age-dependent response. Proc Natl Acad Sci USA 88:9873–9877, 1991Google Scholar
  40. 40.
    Reiser KM: Nonenzymatic glycation of collagen in aging and diabetes. Proc Soc Exp Biol Med 218:23–37, 1998Google Scholar
  41. 41.
    Berlett BS, Stadtman ER: Protein oxidation in aging, disease, and oxidative stress. J Biol Chem 272:20313–20316, 1997Google Scholar
  42. 42.
    Rattan SI: Synthesis, modifications, and turnover of proteins during aging. Exp Gerontol 31:33–47, 1996Google Scholar
  43. 43.
    Cabiscol E, Levine RL: Carbonic anhydrase III. Oxidative modi-fication in vivo and loss of phosphatase activity during aging. J Biol Chem 270:14742–14747, 1995Google Scholar
  44. 44.
    Zuniga A, Gafni A: Age-related modifications in rat cardiac phosphoglycerate kinase. Rejuvenation of the old enzyme by unfolding-refolding. Biochim Biophys Acta 955:50–57, 1988Google Scholar
  45. 45.
    Bukau B, Horwich AL: The Hsp70 and Hsp60 chaperone machines. Cell 92:351–366, 1998Google Scholar
  46. 46.
    Muller K, Gram J, Bollerslev J, Diamant M, Barington T, Hansen MB, Bendtzen K: Down-regulation of monocyte function by treatment of healthy adults with 1α, 25 dihydroxyvitamin D3. Int J Immunopharmacol 13:525–530, 1991Google Scholar
  47. 47.
    Feder ME, Hofmann GE: Heat-shock proteins, molecular chaperones, and the stress response: evolutionary and ecological physiology. Annu Rev Physiol 61:243–282, 1999Google Scholar
  48. 48.
    Heydari AR, Takahashi R, Gutsmann A, You S, Richardson A: Hsp70 and aging. Experientia 50:1092–1098, 1994Google Scholar
  49. 49.
    Blake MJ, Fargnoli J, Gershon D, Holbrook NJ: Concomitant decline in heat-induced hyperthermia and HSP70 mRNA expression in aged rats. Am J Physiol 260:R663–R667, 1991Google Scholar
  50. 50.
    Wu B, Gu MJ, Heydari AR, Richardson A: The effect of age on the synthesis of two heat shock proteins in the hsp70 family. J Gerontol 48:B50–B56, 1993Google Scholar
  51. 51.
    Liu AY, Lin Z, Choi HS, Sorhage F, Li B: Attenuated induction of heat shock gene expression in aging diploid fibroblasts. J Biol Chem 264:12037–12045, 1989Google Scholar
  52. 52.
    Cristofalo VJ, Doggett DL, Brooks-Frederich KM, Phillips PD: Growth factors as probes of cell aging. Exp Gerontol 24:367–374, 1989Google Scholar
  53. 53.
    Miyaishi O, Ito Y, Kozaki K, Sato T, Takechi H, Nagata K, Saga S: Age-related attenuation of HSP47 heat response in fibroblasts. Mech Aging Dev 77:213–226, 1995Google Scholar
  54. 54.
    Gutsmann-Conrad A, Heydari AR, You S, Richardson A: The expression of heat shock protein 70 decreases with cellular senescence in vitro and in cells derived from young and old human subjects. Exp Cell Res 241:404–413, 1998Google Scholar
  55. 55.
    Fargnoli J, Kunisada T, Fornace AJ Jr, Schneider EL, Holbrook NJ: Decreased expression of heat shock protein 70 mRNA and protein after heat treatment in cells of aged rats. Proc Natl Acad Sci USA 87:846–850, 1990Google Scholar
  56. 56.
    Pahlavani MA, Harris MD, Moore SA, Weindruch R, Richardson A: The expression of heat shock protein 70 decreases with age in lymphocytes from rats and rhesus monkeys. Exp Cell Res 218: 310–318, 1995Google Scholar
  57. 57.
    Locke M, Tanguay RM: Diminished heat shock response in the aged myocardium. Cell Stress Chaperones 1:251–260, 1996Google Scholar
  58. 58.
    Choi HS, Lin Z, Li BS, Liu AY: Age-dependent decrease in the heat-inducible DNA sequence-specific binding activity in human diploid fibroblasts. J Biol Chem 265:18005–18011, 1990Google Scholar
  59. 59.
    Effros RB, Zhu X, Walford RL: Stress response of senescent T lymphocytes: Reduced hsp70 is independent of the proliferative block. J Gerontol 49:B65–B70, 1994Google Scholar
  60. 60.
    Bonelli MA, Alfieri RR, Petronini PG, Brigotti M, Campanini C, Borghetti AF: Attenuated expression of 70-kDa heat shock protein in WI-38 human fibroblasts during aging in vitro. Exp Cell Res 252:20–32, 1999Google Scholar
  61. 61.
    Bachelet M, Mariethoz E, Banzet N, Souil E, Pinot F, Polla CZ, Durand P, Bouchaert I, Polla BS: Flow cytometry is a rapid and reliable method for evaluating heat shock protein 70 expression in human monocytes. Cell Stress Chaperones 3:168–176, 1998Google Scholar
  62. 62.
    Udelsman R, Blake MJ, Stagg CA, Li DG, Putney DJ, Holbrook NJ: Vascular heat shock protein expression in response to stress. Endocrine and autonomic regulation of this age-dependent response. J Clin Invest 91:465–473, 1993Google Scholar
  63. 63.
    Fehrenbach E, Passek F, Niess AM, Pohla H, Weinstock C, Dickhuth HH, Northoff H: HSP expression in human leukocytes is modulated by endurance exercise. Med Sci Sports Exerc 32:592–600, 2000Google Scholar
  64. 64.
    Muramatsu T, Hatoko M, Tada H, Shirai T, Ohnishi T: Age-related decrease in the inductability of heat shock protein 72 in normal human skin. Br J Dermatol 134:1035–1038, 1996Google Scholar
  65. 65.
    Gutsmann-Conrad A, Pahlavani MA, Heydari AR, Richardson A: Expression of heat shock protein 70 decreases with age in hepatocytes and splenocytes from female rats. Mech Aging Dev 107:255–270, 1999Google Scholar
  66. 66.
    Unno K, Asakura H, Shibuya Y, Kaiho M, Okada S, Oku N: Increase in basal level of Hsp70, consisting chiefly of constitutively expressed Hsp70 (Hsc70) in aged rat brain. J Gerontol A Biol Sci Med Sci 55:B329–B335, 2000Google Scholar
  67. 67.
    Gandour-Edwards R, McClaren M, Isseroff RR: Immunolocalization of low-molecular-weight stress protein HSP 27 in normal skin and common cutaneous lesions. Am J Dermatopathol 16:504–509, 1994Google Scholar
  68. 68.
    Sphund S, Gershon D: Alterations in the chaperone activity of hsp 70 in aging organisms. Arch. Gerontol 24:125–132, 1997Google Scholar
  69. 69.
    Friedman A, Beller DI: The effect of adherence on the in vitro induction of cytocidal activity by macrophages. Immunology 61:469–474, 1987Google Scholar
  70. 70.
    Sporn SA, Eierman DF, Johnson CE, Morris J, Martin G, Ladner M, Haskill S: Monocyte adherence results in selective induction of novel genes sharing homology with mediators of inflammation and tissue repair. J Immunol 144:4434–4441, 1990Google Scholar
  71. 71.
    Clerget M, Polla BS: Erythrophagocytosis induces heat shock protein synthesis by human monocytes-macrophages. Proc Natl Acad Sci USA 87:1081–1085, 1990Google Scholar
  72. 72.
    Flohe S, Dominguez Fernandez E, Ackermann M, Hirsch T, Borgermann J, Schade FU: Endotoxin tolerance in rats: Expression of TNF-alpha, IL-6, IL-10, VCAM-1 AND HSP 70 in lung and liver during endotoxin shock. Cytokine 11:796–804, 1999Google Scholar
  73. 73.
    Eizirik DL, Welsh M, Strandell E, Welsh N, Sandler S: Interleukin-1 beta depletes insulin messenger ribonucleic acid and increases the heat shock protein hsp70 in mouse pancreatic islets without impairing the glucose metabolism. Endocrinology 127: 2290–2297, 1990Google Scholar
  74. 74.
    Margulis BA, Sandler S, Eizirik DL, Welsh N, Welsh M: Liposomal delivery of purified heat shock protein hsp70 into rat pancreatic islets as protection against interleukin 1 beta-induced impaired beta-cell function. Diabetes 40:1418–1422, 1991Google Scholar
  75. 75.
    Housby JN, Cahill CM, Chu B, Prevelige R, Bickford K, Stevenson MA, Calderwood SK: Non-steroidal anti-inflammatory drugs inhibit the expression of cytokines and induce HSP70 in human monocytes. Cytokine 11:347–358, 1999Google Scholar
  76. 76.
    Cahill CM, Waterman WR, Xie Y, Auron PE, Calderwood SK: Transcriptional repression of the prointerleukin 1beta gene by heat shock factor 1. J Biol Chem 271:24874–24879, 1996Google Scholar
  77. 77.
    Singh IS, Viscardi RM, Kalvakolanu I, Calderwood S, Hasday JD: Inhibition of tumor necrosis factor-α transcription in macrophages exposed to febrile range temperature. A possible role for heat shock factor-1 as a negative transcriptional regulator. J Biol Chem 275:9841–9848, 2000Google Scholar
  78. 78.
    Singh IS, He JR, Calderwood S, Hasday JD: A high affinity HSF-1 binding site in the 5′-untranslated region of the murine tumor necrosis factor-α gene is a transcriptional repressor. J Biol Chem 277:4981–4988, 2002Google Scholar
  79. 79.
    Stephanou A, Isenberg DA, Akira S, Kishimoto T, Latchman DS: The nuclear factor interleukin-6 (NF-IL6) and signal transducer and activator of transcription-3 (STAT-3) signalling pathways co-operate to mediate the activation of the hsp90β gene by inteleukin-6 but have opposite effects on its inducibility by heat shock. Biochem J 330:189–195, 1998Google Scholar
  80. 80.
    Stephanou A, Isenberg DA, Nakajima K, Latchman DS: Signal transducer and activator of transcription-1 and heat shock factor-1 interact and activate the transcription of the Hsp-70 an Hsp-90β gene promoters. J Biol Chem 274:1723–1728, 1999Google Scholar
  81. 81.
    Dai R, Frejtag W, He B, Zhang Y, Mivechi NF: c-Jun NH2-terminal kinase targeting and phosphorylation of heat shock factor-1 suppress its transcriptional activity. J Biol Chem 275: 18210–18218, 2000Google Scholar
  82. 82.
    Straub RH, Konecna L, Hrach S, Rothe G, Kreutz M, Scholmerich J, Falk W, Lang B: Serum dehydroepiandrosterone (DHEA) and DHEA sulfate are negatively correlated with serum interleukin-6 (IL-6), and DHEA inhibits IL-6 secretion from mononuclear cells in man in vitro: Possible link between endocrinosenescence and immunosenescence. J Clin Endocrinol Metab 83:2012–2017, 1998Google Scholar
  83. 83.
    Ershler WB: Interleukin-6: a cytokine for gerontologists. J Am Geriatr Soc 41:176–181, 1993wGoogle Scholar

Copyright information

© Plenum Publishing Corporation 2002

Authors and Affiliations

  • R. Njemini
    • 1
  • M. Vanden Abeele
    • 1
  • C. Demanet
    • 2
  • M. Lambert
    • 1
  • S. Vandebosch
    • 1
  • T. Mets
    • 1
    Email author
  1. 1.Geriatric Unit, Academic HospitalFree University Brussels (VUB)Belgium
  2. 2.HLA and Immunology Laboratory, Academic HospitalFree University Brussels (VUB)Belgium

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