Skip to main content
SpringerLink
Log in

The regulation of TNFα production after heat and endotoxin stimulation is dependent on Annexin-A1 and HSP70

  • Original Paper
  • Published:
Cell Stress and Chaperones Aims and scope

Abstract

Febrile temperatures can induce stress responses which protect cells from damage and can reduce inflammation during infections and sepsis. However, the mechanisms behind the protective functions of heat in response to the bacterial endotoxin LPS are unclear. We have recently shown that Annexin-1 (ANXA1)-deficient macrophages exhibited higher TNFα levels after LPS stimulation. Moreover, we have previously reported that ANXA1 can function as a stress protein. Therefore in this study, we determined if ANXA1 is involved in the protective effects of heat on cytokine levels in macrophages after heat and LPS. Exposure of macrophages to 42 °C for 1 h prior to LPS results in an inhibition of TNFα production, which was not evident in ANXA1−/− macrophages. We show that this regulation involves primarily MYD88-independent pathways. ANXA1 regulates TNFα mRNA stability after heat and LPS, and this is dependent on endogenous ANXA1 expression and not exogenously secreted factors. Further mechanistic studies revealed the possible involvement of the heat shock protein HSP70 and JNK in the heat and inflammatory stress response regulated by ANXA1. This study shows that ANXA1, an immunomodulatory protein, is critical in the heat stress response induced after heat and endotoxin stimulation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

Abbreviations

LPS:

Lipopolysaccharide

TNF:

Tumor necrosis factor

IL:

Interleukin

TRIF:

TIR-domain-containing adapter-inducing interferon-β

ANXA1:

Annexin-A1

MYD88:

Myeloid differentiation primary response 88

HSP:

Heat shock protein

JNK:

Jun kinase

ERK:

Extracellular signal-regulated kinase

MAPK:

Mitogen-activated protein kinase

MKP1:

Mitogen-activated protein kinase phosphatase-1

References

  • Akira S, Takeda K, Kaisho T (2001) Toll-like receptors: critical proteins linking innate and acquired immunity. Nat Immunol 2:675–680. doi:10.1038/90609

    Article  CAS  PubMed  Google Scholar 

  • Alldridge LC, Harris HJ, Plevin R, Hannon R, Bryant CE (1999) The annexin protein lipocortin 1 regulates the MAPK/ERK pathway. J Biol Chem 274:37620–37628

    Article  CAS  PubMed  Google Scholar 

  • Asea A et al (2002) Novel signal transduction pathway utilized by extracellular HSP70: role of toll-like receptor (TLR) 2 and TLR4. J Biol Chem 277:15028–15034. doi:10.1074/jbc.M200497200

    Article  CAS  PubMed  Google Scholar 

  • Bist P et al (2013) Annexin-A1 regulates TLR-mediated IFN-beta production through an interaction with TANK-binding kinase 1. J Immunol 191:4375–4382. doi:10.4049/jimmunol.1301504

    Article  CAS  PubMed  Google Scholar 

  • Biswas SK, Lopez-Collazo E (2009) Endotoxin tolerance: new mechanisms, molecules and clinical significance. Trends Immunol 30:475–487. doi:10.1016/j.it.2009.07.009

    Article  CAS  PubMed  Google Scholar 

  • Camps M, Nichols A, Arkinstall S (2000) Dual specificity phosphatases: a gene family for control of MAP kinase function. FASEB J 14:6–16

    CAS  PubMed  Google Scholar 

  • Cargnello M, Roux PP (2011) Activation and function of the MAPKs and their substrates, the MAPK-activated protein kinases. Microbiol Mol Biol Rev 75:50–83. doi:10.1128/MMBR. 00031-10

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Castro-Caldas M, Duarte CB, Carvalho AP, Lopes MC (2002) Dexamethasone induces the secretion of annexin I in immature lymphoblastic cells by a calcium-dependent mechanism. Mol Cell Biochem 237:31–38

    Article  CAS  PubMed  Google Scholar 

  • Chi H, Barry SP, Roth RJ, Wu JJ, Jones EA, Bennett AM, Flavell RA (2006) Dynamic regulation of pro- and anti-inflammatory cytokines by MAPK phosphatase 1 (MKP-1) in innate immune responses. Proc Natl Acad Sci U S A 103:2274–2279. doi:10.1073/pnas.0510965103

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Cooper ZA, Singh IS, Hasday JD (2010) Febrile range temperature represses TNF-alpha gene expression in LPS-stimulated macrophages by selectively blocking recruitment of Sp1 to the TNF-alpha promoter. Cell Stress Chaperones 15:665–673. doi:10.1007/s12192-010-0179-9

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • English J, Pearson G, Wilsbacher J, Swantek J, Karandikar M, Xu S, Cobb MH (1999) New insights into the control of MAP kinase pathways. Exp Cell Res 253:255–270. doi:10.1006/excr.1999.4687

    Article  CAS  PubMed  Google Scholar 

  • Ensor JE, Wiener SM, McCrea KA, Viscardi RM, Crawford EK, Hasday JD (1994) Differential effects of hyperthermia on macrophage interleukin-6 and tumor necrosis factor-alpha expression. Am J Physiol 266:C967–974

    CAS  PubMed  Google Scholar 

  • Gabai VL, Meriin AB, Mosser DD, Caron AW, Rits S, Shifrin VI, Sherman MY (1997) Hsp70 prevents activation of stress kinases. A novel pathway of cellular thermotolerance. J Biol Chem 272:18033–18037

    Article  CAS  PubMed  Google Scholar 

  • Garekar S, Heidemann SM, Glibetic M (2006) Heat stress response results in increased macrophage inflammatory protein-2 concentration in a lipopolysaccharide-exposed macrophage cell line. J Endotoxin Res 12:87–92. doi:10.1179/096805106X89071

    Article  CAS  PubMed  Google Scholar 

  • Gupta A, Cooper ZA, Tulapurkar ME, Potla R, Maity T, Hasday JD, Singh IS (2013) Toll-like receptor agonists and febrile range hyperthermia synergize to induce heat shock protein 70 expression and extracellular release. J Biol Chem 288:2756–2766. doi:10.1074/jbc.M112.427336

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Hoebe K et al (2003) Identification of Lps2 as a key transducer of MyD88-independent TIR signalling. Nature 424:743–748. doi:10.1038/nature01889

    Article  CAS  PubMed  Google Scholar 

  • Hunt C, Calderwood S (1990) Characterization and sequence of a mouse hsp70 gene and its expression in mouse cell lines. Genetics 87:199–204

    CAS  Google Scholar 

  • Kim GY, Lee HB, Lee SO, Rhee HJ, Na DS (1997) Chaperone-like function of lipocortin 1. Biochem Mol Biol Int 43:521–528

    CAS  PubMed  Google Scholar 

  • Kyriakis JM, Avruch J (2001) Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress and inflammation. Physiol Rev 81:807–869

    CAS  PubMed  Google Scholar 

  • Kyriakis JM et al (1994) The stress-activated protein kinase subfamily of c-Jun kinases. Nature 369:156–160. doi:10.1038/369156a0

    Article  CAS  PubMed  Google Scholar 

  • Lee KH, Lee CT, Kim YW, Han SK, Shim YS, Yoo CG (2005) Preheating accelerates mitogen-activated protein (MAP) kinase inactivation post-heat shock via a heat shock protein 70-mediated increase in phosphorylated MAP kinase phosphatase-1. J Biol Chem 280:13179–13186. doi:10.1074/jbc.M410059200

    Article  CAS  PubMed  Google Scholar 

  • Li CL, Wang XY, Shao J, Zhang JS, Feng WG, Wang YB, Chang ZL (2001) Heat shock inhibits IL-12 p40 expression through NF-kappa B signalling pathway in murine macrophages. Cytokine 16:153–159. doi:10.1006/cyto.2001.0971

    Article  CAS  PubMed  Google Scholar 

  • Lim LH, Pervaiz S (2007) Annexin 1: the new face of an old molecule. FASEB J 21:968–975. doi:10.1096/fj.06-7464rev

    Article  CAS  PubMed  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25:402–408. doi:10.1006/meth.2001.1262

    Article  CAS  PubMed  Google Scholar 

  • Meng X, Harken AH (2002) The interaction between Hsp70 and TNF-alpha expression: a novel mechanism for protection of the myocardium against post-injury depression. Shock 17:345–353

    Article  PubMed  Google Scholar 

  • Morimoto RI (1998) Regulation of the heat shock transcriptional response: cross talk between a family of heat shock factors, molecular chaperones, and negative regulators. Genes & Development 12:3788–3796. doi:10.1101/gad.12.24.3788

    Article  CAS  Google Scholar 

  • Nair S, Hande MP, Lim LH (2010) Annexin-1 protects MCF7 breast cancer cells against heat-induced growth arrest and DNA damage. Cancer Lett 294:111–117. doi:10.1016/j.canlet.2010.01.026

    Article  CAS  PubMed  Google Scholar 

  • Perretti M, Croxtall JD, Wheller SK, Goulding NJ, Hannon R, Flower RJ (1996) Mobilizing lipocortin 1 in adherent human leukocytes downregulates their transmigration. Nat Med 2:1259–1262

    Article  CAS  PubMed  Google Scholar 

  • Rattan SI, Eskildsen-Helmond YE, Beedholm R (2004) Molecular mechanisms of anti-aging hormetic effects of mild heat stress on human cells. Nonlinearity Biol Toxicol Med 2:105–116. doi:10.1080/15401420490464376

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Raynal P, Pollard HB (1994) Annexins: the problem of assessing the biological role for a gene family of multifunctional calcium- and phospholipid-binding proteins. Biochim Biophys Acta 1197:63–93

    Article  CAS  PubMed  Google Scholar 

  • Rhee HJ, Kim GY, Huh JW, Kim SW, Na DS (2000) Annexin I is a stress protein induced by heat, oxidative stress and a sulfhydryl-reactive agent. Eur J Biochem 267:3220–3225

    Article  CAS  PubMed  Google Scholar 

  • Shi Y et al (2006) The inhibition of LPS-induced production of inflammatory cytokines by HSP70 involves inactivation of the NF-kappaB pathway but not the MAPK pathways. Shock 26:277–284. doi:10.1097/01.shk.0000223134.17877.ad

    Article  CAS  PubMed  Google Scholar 

  • Snyder YM, Guthrie L, Evans GF, Zuckerman SH (1992) Transcriptional inhibition of endotoxin-induced monokine synthesis following heat shock in murine peritoneal macrophages. J Leukoc Biol 51:181–187

    CAS  PubMed  Google Scholar 

  • Takeda K, Akira S (2004) TLR signaling pathways. Semin Immunol 16:3–9. doi:10.1016/j.smim.2003.10.003

    Article  CAS  PubMed  Google Scholar 

  • Toshchakov V et al (2002) TLR4, but not TLR2, mediates IFN-beta-induced STAT1alpha/beta-dependent gene expression in macrophages. Nat Immunol 3:392–398. doi:10.1038/ni774

    Article  CAS  PubMed  Google Scholar 

  • van der Bruggen T, Nijenhuis S, van Raaij E, Verhoef J, van Asbeck BS (1999) Lipopolysaccharide-induced tumor necrosis factor alpha production by human monocytes involves the raf-1/MEK1-MEK2/ERK1-ERK2 pathway. Infect Immun 67:3824–3829

    PubMed Central  PubMed  Google Scholar 

  • Yang YH et al (2006) Annexin 1 negatively regulates IL-6 expression via effects on p38 MAPK and MAPK phosphatase-1. J Immunol 177:8148–8153

    Article  CAS  PubMed  Google Scholar 

  • Yu H, Sun Y, Haycraft C, Palanisamy V, Kirkwood KL (2011) MKP-1 regulates cytokine mRNA stability through selectively modulation subcellular translocation of AUF1. Cytokine 56:245–255

    Article  CAS  PubMed Central  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from Biomedical Research Council (BMRC) of Singapore (07/1/21/19/507) and NUHS (R-185-000-239-750) to LL.

Conflict of interest

The authors declare no conflict of interest.

Author contributions

SN and LL conceived and designed the experiments; SN, SA, and LHL performed the experiments; SN and LL analyzed the data; JYL contributed reagents/tools; and SN and LL wrote the manuscript.

Author information

Authors and Affiliations

  1. Department of Physiology, Yong Loo Lin School of Medicine, National University Health System, National University of Singapore, 28 Medical Drive, Singapore, 117456, Singapore

    Sunitha Nair, Suruchi Arora, Lay Hoon Lee & Lina H.K. Lim

  2. NUS Immunology Program, Life Sciences Institute, National University of Singapore, Singapore, 117456, Singapore

    Sunitha Nair, Suruchi Arora, Lay Hoon Lee & Lina H.K. Lim

  3. Singapore Immunology Network (SigN), 8A Biomedical Grove, Immunos, Singapore, 138648, Singapore

    Jyue Yuan Lim

Authors
  1. Sunitha Nair
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Suruchi Arora
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jyue Yuan Lim
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Lay Hoon Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Lina H.K. Lim
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lina H.K. Lim.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Nair, S., Arora, S., Lim, J. et al. The regulation of TNFα production after heat and endotoxin stimulation is dependent on Annexin-A1 and HSP70. Cell Stress and Chaperones 20, 583–593 (2015). https://doi.org/10.1007/s12192-015-0580-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12192-015-0580-5

Keywords

Search

Navigation