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Features of Stress-Induced Changes of HSP70 Expression in Populations of Immunocompetent Cells

  • Anna A. Boyko
  • Natalya I. Troyanova
  • Julia D. Teterina
  • Tatyana L. Azhikina
  • Sergey S. Vetchinin
  • Elena I. Kovalenko
  • Alexander M. SapozhnikovEmail author
Chapter
Part of the Heat Shock Proteins book series (HESP, volume 15)

Abstract

The results of intracellular protein measurement carrying out with the use of monoclonal antibodies, strongly depend on the localization of antibody binding epitopes in the structure of the molecular target. Our previous study has shown pronounced differences in the pattern of cellular stress response detected by monitoring of intracellular HSP70 content, which was assessed using flow cytometry and monoclonal antibodies interacting with various domains of these molecules. In the present work we investigate the features of stress-induced changes in the expression of HSP70 in populations of mononuclear and granulocytic cells from human peripheral blood using standard methods: PCR, Western-blot and flow-cytometric analysis. A particular feature of this study is an application of a number of antibodies including monoclonal antibodies specific for the total pool of HSP70, separately for the inducible or constitutive form of this protein, as well as for the different domains of the HSP70 molecule. The results provide new additional information on HSP70-related stress-induced processes in populations of immunocompetent cells.

Keywords

Anti-HSP70 antibodies Heat shock Heat shock proteins HSP70 expression Human leukocytes 

Abbreviations

HS

Heat shock

Hsc70

Constitutive form of HSP70

HSP

Heat shock proteins

HSP70

Heat shock protein 70 kDa

Hsp70

Inducible form of HSP70

MFI

Means of fluorescence intensity

PBMC

Peripheral blood mononuclear cells

PMN

Polymorph nuclear leukocytes

Notes

Acknowledgements

This work was supported by Russian Science Foundation, grant # 16-15-10404.

References

  1. Angelidis CE, Lazaridis I, Pagoulatos GN (1999) Aggregation of hsp70 and hsc70 in vivo is distinct and temperature- dependent and their chaperone function is directly related to non-aggregated forms. Eur J Biochem 259:505–512CrossRefGoogle Scholar
  2. Boyko AA, Vetchinin SS, Sapozhnikov AM, Kovalenko EI (2014) Changes in the heat shock 70 kDa protein level in human neutrophils induced by heat shock. Russ J Bioorg Chem 40:488–498CrossRefGoogle Scholar
  3. Boyko AA, Azhikina TL, Streltsova MA, Sapozhnikov AM, Kovalenko EI (2017) HSP70 in human polymorphonuclear and mononuclear leukocytes: comparison of the protein content and transcriptional activity of HSPA genes. Cell Stress and Chaperones 22:67–76CrossRefGoogle Scholar
  4. Calderwood SK, Murshid A, Prince T (2009) The shock of aging: molecular chaperones and the heat shock response in longevity and aging. Gerontology 55:550–558CrossRefGoogle Scholar
  5. Cuanalo-Contreras K, Mukherjee A, Soto C (2013) Role of protein misfolding and proteostasis deficiency in protein misfolding diseases and aging. Int J Cell Biol 2013:638083CrossRefGoogle Scholar
  6. Eid NS, Kravath RE, Lanks KW (1987) Heat-shock protein synthesis by human polymorphonuclear cells. J Exp Med 165:1448–1452CrossRefGoogle Scholar
  7. Ferat-Osorio E, Sánchez-Anaya A, Gutiérrez-Mendoza M, Boscó-Gárate I, Wong-Baeza I, Pastelin-Palacios R, Pedraza-Alva G, Bonifaz LC, Cortés-Reynosa P, Pérez-Salazar E, Arriaga-Pizano L, López-Macías C, Rosenstein Y, Isibasi A (2014) Heat shock protein 70 down-regulates the production of toll-like receptor-induced pro-inflammatory cytokines by a heat shock factor-1/constitutive heat shock element-binding factor-dependent mechanism. J Inflamm (Lond) 11:19CrossRefGoogle Scholar
  8. Kovalenko EI, Boyko AA, Semenkov VF, Lutsenko GV, Grechikhina MV, Kanevskiy LM, Azhikina TL, Telford WG, Sapozhnikov AM (2014) ROS production, intracellular HSP70 levels and their relationship in human neutrophils: effects of age. Oncotarget 5:11800–11812CrossRefGoogle Scholar
  9. Mathur SK, Sistonen L, Brown IR, Murphy SP, Sarge KD, Morimoto RI (1994) Deficient induction of human hsp70 heat shock gene transcription in Y79 retinoblastoma cells despite activation of heat shock factor 1. Proc Natl Acad Sci U S A 91:8695–8699CrossRefGoogle Scholar
  10. McLeod IX, Jia W, He YW (2012) The contribution of autophagy to lymphocyte survival and homeostasis. Immunol Rev 249:195–204CrossRefGoogle Scholar
  11. Multhoff G (2007) Heat shock protein 70 (Hsp70): membrane location, export and immunological relevance. Methods 43:229–237CrossRefGoogle Scholar
  12. Muralidharan S, Mandrekar P (2013) Cellular stress response and innate immune signaling: integrating pathways in host defense and inflammation. J Leukoc Biol 94:1167–1184CrossRefGoogle Scholar
  13. Njemini R, Abeele MV, Demanet C, Lambert M, Vandebosch S, Mets T (2002) Age-related decrease in the inducibility of heat-shock protein 70 in human peripheral blood mononuclear cells. J Clin Immunol 22:195–205CrossRefGoogle Scholar
  14. Polla BS, Stubbe H, Kantengwa S, Maridonneau-Parini I, Jacquier-Sarlin MR (1995) Differential induction of stress proteins and functional effects of heat shock in human phagocytes. Inflammation 19:363–378CrossRefGoogle Scholar
  15. Schneider EM, Niess AM, Lorenze I, Northoff H, Fehrenbach E (2002) Inducible hsp70 expression analysis after heat and physical exercise: transcriptional, protein expression, and subcellular localization. Ann N Y Acad Sci 973:8–12CrossRefGoogle Scholar
  16. Tan H, Xu Y, Xu J, Wang F, Nie S, Yang M, Yuan J, Tanguay RM, Wu T (2007) Association of increased heat shock protein 70 levels in the lymphocyte with high risk of adverse pregnancy outcomes in early pregnancy: a nested case-control study. Cell Stress Chaperones 12:230–236CrossRefGoogle Scholar
  17. Theodorakis NG, Morimoto RI (1987) Posttranscriptional regulation of hsp70 expression in human cells: effects of heat shock, inhibition of protein synthesis, and adenovirus infection on translation and mRNA stability. Mol Cell Biol 7(12):4357–4368CrossRefGoogle Scholar
  18. Wang S, Diller KR, Aggarwal SJ (2003) Kinetics study of endogenous heat shock protein 70 expression. J Biomech Eng 125:794–797CrossRefGoogle Scholar
  19. Zhang X, Kluger Y, Nakayama Y, Poddar R, Whitney C, DeTora A, Weissman SM, Newburger PE (2004) Gene expression in mature neutrophils: early responses to inflammatory stimuli. J Leukoc Biol 75:358–372CrossRefGoogle Scholar
  20. Zhuravleva A, Clerico EM, Gierasch LM (2012) An interdomain energetic tug-of-war creates the allosterically active state in Hsp70 molecular chaperones. Cell 151:1296–1307CrossRefGoogle Scholar

Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  • Anna A. Boyko
    • 1
  • Natalya I. Troyanova
    • 1
  • Julia D. Teterina
    • 1
  • Tatyana L. Azhikina
    • 1
  • Sergey S. Vetchinin
    • 2
  • Elena I. Kovalenko
    • 1
  • Alexander M. Sapozhnikov
    • 1
    Email author
  1. 1.Laboratory of Cell Interactions, Department of ImmunologyShemyakin - Ovchinnikov Institute of Bioorganic ChemistryMoscowRussia
  2. 2.The State Research Center for Applied Microbiology and BiotechnologyObolenskRussia

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