Cell Stress and Chaperones

, Volume 23, Issue 3, pp 373–384 | Cite as

Extracellular Hsp70 induces inflammation and modulates LPS/LTA-stimulated inflammatory response in THP-1 cells

  • Andrea Hulina
  • Marija Grdić Rajković
  • Daniela Jakšić Despot
  • Dubravko Jelić
  • Ana Dojder
  • Ivana Čepelak
  • Lada Rumora
Original Paper
  • 240 Downloads

Abstract

Extracellular Hsp70 (eHsp70) can act as damage-associated molecular pattern (DAMP) via Toll-like receptors TLR2 and TLR4, and stimulate immune and inflammatory responses leading to sterile inflammation and propagation of already existing inflammation. It was found elevated in the blood of patients with chronic obstructive pulmonary disease (COPD), who might suffer occasional bacterial colonizations and infections. We used a monocytic THP-1 cell line as a cellular model of systemic compartment of COPD to assess inflammatory effects of eHsp70 when present alone or together with bacterial products lypopolysaccharide (LPS) and lypoteichoic acid (LTA). THP-1 cells were differentiated into macrophage-like cells and treated with various concentrations of recombinant human Hsp70 protein (rhHsp70), LPS (TLR4 agonist), LTA (TLR2 agonist), and their combinations for 4, 12, 24, and 48 h. Concentrations of IL-1α, IL-6, IL-8, and TNF-α were determined by ELISA. Cell viability was assessed by MTS assay, and mode of cell death by luminometric measurements of caspases-3/7, -8, and -9 activities. rhHsp70 showed cell protecting effect by suppressing caspases-3/7 activation, while LPS provoked cytotoxicity through caspases-8 and -3/7 pathway. Regarding inflammatory processes, rhHsp70 alone induced secretion of IL-1α and IL-8, but had modulatory effects on release of all four cytokines when applied together with LPS or LTA. Combined effect with LPS was mainly synergistic, and with LTA mainly antagonistic, although it was cytokine- and time-dependent. Our results confirmed pro-inflammatory function of extracellular Hsp70, and suggest its possible implication in COPD exacerbations caused by bacterial infection through desensitization or inappropriate activation of TLR2 and TLR4 receptors.

Keywords

THP-1 cells Extracellular Hsp70 LPS LTA COPD 

Notes

Acknowledgements

This work has been fully supported by the Croatian Science Foundation under the project number IP-2014-09-1247.

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Copyright information

© Cell Stress Society International 2017

Authors and Affiliations

  1. 1.Department of Medical Biochemistry and Hematology, Faculty of Pharmacy and BiochemistryUniversity of ZagrebZagrebCroatia
  2. 2.Department of Microbiology, Faculty of Pharmacy and BiochemistryUniversity of ZagrebZagrebCroatia
  3. 3.Fidelta d.o.oZagrebCroatia

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