Spandidos Publications Logo
  • About
    • About Spandidos
    • Aims and Scopes
    • Abstracting and Indexing
    • Editorial Policies
    • Reprints and Permissions
    • Job Opportunities
    • Terms and Conditions
    • Contact
  • Journals
    • All Journals
    • Oncology Letters
      • Oncology Letters
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Oncology
      • International Journal of Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular and Clinical Oncology
      • Molecular and Clinical Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Experimental and Therapeutic Medicine
      • Experimental and Therapeutic Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Molecular Medicine
      • International Journal of Molecular Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Biomedical Reports
      • Biomedical Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Oncology Reports
      • Oncology Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular Medicine Reports
      • Molecular Medicine Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • World Academy of Sciences Journal
      • World Academy of Sciences Journal
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Functional Nutrition
      • International Journal of Functional Nutrition
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Epigenetics
      • International Journal of Epigenetics
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Medicine International
      • Medicine International
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
  • Articles
  • Information
    • Information for Authors
    • Information for Reviewers
    • Information for Librarians
    • Information for Advertisers
    • Conferences
  • Language Editing
Spandidos Publications Logo
  • About
    • About Spandidos
    • Aims and Scopes
    • Abstracting and Indexing
    • Editorial Policies
    • Reprints and Permissions
    • Job Opportunities
    • Terms and Conditions
    • Contact
  • Journals
    • All Journals
    • Biomedical Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Experimental and Therapeutic Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Epigenetics
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Functional Nutrition
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Molecular Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Medicine International
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular and Clinical Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular Medicine Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Oncology Letters
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Oncology Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • World Academy of Sciences Journal
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
  • Articles
  • Information
    • For Authors
    • For Reviewers
    • For Librarians
    • For Advertisers
    • Conferences
  • Language Editing
Login Register Submit
  • This site uses cookies
  • You can change your cookie settings at any time by following the instructions in our Cookie Policy. To find out more, you may read our Privacy Policy.

    I agree
Search articles by DOI, keyword, author or affiliation
Search
Advanced Search
presentation
Molecular Medicine Reports
Join Editorial Board Propose a Special Issue
Print ISSN: 1791-2997 Online ISSN: 1791-3004
Journal Cover
July-2018 Volume 18 Issue 1

Full Size Image

Sign up for eToc alerts
Recommend to Library

Journals

International Journal of Molecular Medicine

International Journal of Molecular Medicine

International Journal of Molecular Medicine is an international journal devoted to molecular mechanisms of human disease.

International Journal of Oncology

International Journal of Oncology

International Journal of Oncology is an international journal devoted to oncology research and cancer treatment.

Molecular Medicine Reports

Molecular Medicine Reports

Covers molecular medicine topics such as pharmacology, pathology, genetics, neuroscience, infectious diseases, molecular cardiology, and molecular surgery.

Oncology Reports

Oncology Reports

Oncology Reports is an international journal devoted to fundamental and applied research in Oncology.

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine is an international journal devoted to laboratory and clinical medicine.

Oncology Letters

Oncology Letters

Oncology Letters is an international journal devoted to Experimental and Clinical Oncology.

Biomedical Reports

Biomedical Reports

Explores a wide range of biological and medical fields, including pharmacology, genetics, microbiology, neuroscience, and molecular cardiology.

Molecular and Clinical Oncology

Molecular and Clinical Oncology

International journal addressing all aspects of oncology research, from tumorigenesis and oncogenes to chemotherapy and metastasis.

World Academy of Sciences Journal

World Academy of Sciences Journal

Multidisciplinary open-access journal spanning biochemistry, genetics, neuroscience, environmental health, and synthetic biology.

International Journal of Functional Nutrition

International Journal of Functional Nutrition

Open-access journal combining biochemistry, pharmacology, immunology, and genetics to advance health through functional nutrition.

International Journal of Epigenetics

International Journal of Epigenetics

Publishes open-access research on using epigenetics to advance understanding and treatment of human disease.

Medicine International

Medicine International

An International Open Access Journal Devoted to General Medicine.

Journal Cover
July-2018 Volume 18 Issue 1

Full Size Image

Sign up for eToc alerts
Recommend to Library

  • Article
  • Citations
    • Cite This Article
    • Download Citation
    • Create Citation Alert
    • Remove Citation Alert
    • Cited By
  • Similar Articles
    • Related Articles (in Spandidos Publications)
    • Similar Articles (Google Scholar)
    • Similar Articles (PubMed)
  • Download PDF
  • Download XML
  • View XML
Article

Inhibition of granzyme B activity blocks inflammation induced by lipopolysaccharide through regulation of endoplasmic reticulum stress signaling in NK92 cells

  • Authors:
    • Lei Wang
    • Shaowei Jiang
    • Ling Xiao
    • Lin Chen
    • Yanyan Zhang
    • Jing Tong
  • View Affiliations / Copyright

    Affiliations: Department of Clinical Immunology, Institute of Laboratory Medicine, Hubei University of Chinese Medicine, Wuhan, Hubei 430065, P.R. China, Aristogenesis Genetic Laboratory, Wuhan Children's Hospital (Wuhan Maternal and Child Healthcare Hospital), Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430074, P.R. China
  • Pages: 580-586
    |
    Published online on: May 9, 2018
       https://doi.org/10.3892/mmr.2018.8995
  • Expand metrics +
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )
Cited By (CrossRef): 0 citations Loading Articles...

This article is mentioned in:



Abstract

Granzyme B (GrB) is a serine protease that is expressed in the lytic granules of natural killer (NK) cells and cytotoxic T lymphocytes (CTL), and which has been widely reported to serve a crucial role for target cell apoptosis. GrB may serve a non‑cytotoxic role in inflammation, but the evidence remains unclear. The present study aimed to establish an inflammatory cell model by using NK92 cells stimulated with lipopolysaccharide (LPS) to investigate whether GrB was involved in the development of inflammation. The extracellular levels of tumor necrosis factor‑α (TNF‑α), interleukin‑1β (IL‑1β) and GrB were examined by ELISA, and it was demonstrated that LPS treatment increased the extracellular levels of TNF‑α, IL‑1β and GrB, and these increased expression levels were inhibited by pretreatment with the GrB inhibitor serpin A3N (SA3N). The protein expression levels of glucose‑regulated protein 78 (GRP78), C/EBP homologous protein (CHOP), nuclear factor‑κB (NF‑κB), inhibitor of NF‑κB (IκBα) and GrB were examined by western blot analysis. The results demonstrated that LPS stimulation increased the expression levels of GRP78, CHOP, NF‑κB and GrB, and decreased the expression of IκBα, and these changes were inhibited by SA3N, which indicated that inhibition of GrB activity may suppress endoplasmic reticulum (ER) stress signaling. Therefore, it was suggested that GrB may be a potential pro‑inflammatory factor, and inhibition of GrB activity may aid the prevention of the development of inflammation by suppressing ER stress signaling.
View Figures

Figure 1

Figure 2

Figure 3

Figure 4

View References

1 

Bladergroen BA, Meijer CJ, ten Berge RL, Hack CE, Muris JJ, Dukers DF, Chott A, Kazama Y, Oudejans JJ, Berkum O, et al: Expression of the granzyme B inhibitor, protease inhibitor 9, by tumor cells in patients with non-Hodgkin and Hodgkin lymphoma: A novel protective mechanism for tumor cells to circumvent the immune system? Blood. 99:232–237. 2002. View Article : Google Scholar : PubMed/NCBI

2 

Thomas DA, Scorrano L, Putcha GV, Korsmeyer SJ and Ley TJ: Granzyme B can cause mitochondrial depolarization and cell death in the absence of BID, BAX, and BAK. Proc Natl Acad Sci USA. 98:14985–14990. 2001. View Article : Google Scholar : PubMed/NCBI

3 

Alimonti JB, Shi L, Baijal PK and Greenberg AH: Granzyme B induces BID-mediated cytochrome c release and mitochondrial permeability transition. J Biol Chem. 276:6974–6982. 2001. View Article : Google Scholar : PubMed/NCBI

4 

Mattila JT, Maiello P, Sun T, Via LE and Flynn JL: Granzyme B-expressing neutrophils correlate with bacterial load in granulomas from Mycobacterium tuberculosis-infected cynomolgus macaques. Cell Microbiol. 17:1085–1097. 2015. View Article : Google Scholar : PubMed/NCBI

5 

Classen CF, Bird PI and Debatin KM: Modulation of the granzyme B inhibitor proteinase inhibitor 9 (PI-9) by activation of lymphocytes and monocytes in vitro and by Epstein-Barr virus and bacterial infection. Clin Exp Immunol. 143:534–542. 2006. View Article : Google Scholar : PubMed/NCBI

6 

Rousalova I and Krepela E: Granzyme B-induced apoptosis in cancer cells and its regulation (review). Int J Oncol. 37:1361–1378. 2010.PubMed/NCBI

7 

Lord SJ, Rajotte RV, Korbutt GS and Bleackley RC: Granzyme B: A natural born killer. Immunol Rev. 193:31–38. 2003. View Article : Google Scholar : PubMed/NCBI

8 

Yin XM: Bid, a BH3-only multi-functional molecule, is at the cross road of life and death. Gene. 369:7–19. 2006. View Article : Google Scholar : PubMed/NCBI

9 

Afonina IS, Cullen SP and Martin SJ: Cytotoxic and non-cytotoxic roles of the CTL/NK protease granzyme B. Immunol Rev. 235:105–116. 2010. View Article : Google Scholar : PubMed/NCBI

10 

Froelich CJ, Pardo J and Simon MM: Granule-associated serine proteases: Granzymes might not just be killer proteases. Trends in Immunol. 30:117–123. 2009. View Article : Google Scholar

11 

Afonina IS, Tynan GA, Logue SE, Cullen SP, Bots M, Lüthi AU, Reeves EP, McElvaney NG, Medema JP, Lavelle EC and Martin SJ: Granzyme B-dependent proteolysis acts as a switch to enhance the proinflammatory activity of IL-1α. Mol Cell. 44:265–278. 2011. View Article : Google Scholar : PubMed/NCBI

12 

Kim HJ, Jeong JS, Kim SR, Park SY, Chae HJ and Lee YC: Inhibition of endoplasmic reticulum stress alleviates lipopolysaccharide-induced lung inflammation through modulation of NF-κB/HIF-1α signaling pathway. Sci Rep. 3:11422013. View Article : Google Scholar : PubMed/NCBI

13 

Tjondrokoesoemo A, Schips T, Kanisicak O, Sargent MA and Molkentin JD: Genetic overexpression of Serpina3n attenuates muscular dystrophy in mice. Hum Mol Genet. 25:1192–1202. 2016. View Article : Google Scholar : PubMed/NCBI

14 

Vicuña L, Strochlic DE, Latremoliere A, Bali KK, Simonetti M, Husainie D, Prokosch S, Riva P, Griffin RS, Njoo C, et al: The serine protease inhibitor SerpinA3N attenuates neuropathic pain by inhibiting T cell-derived leukocyte elastase. Nat Med. 21:518–523. 2015. View Article : Google Scholar : PubMed/NCBI

15 

Ang LS, Boivin WA, Williams SJ, Zhao H, Abraham T, Carmine-Simmen K, McManus BM, Bleackley RC and Granville DJ: Serpina3n attenuates granzyme B-mediated decorin cleavage and rupture in a murine model of aortic aneurysm. Cell Death Dis. 2:e2092011. View Article : Google Scholar : PubMed/NCBI

16 

Haile Y, Carmine-Simmen K, Olechowski C, Kerr B, Bleackley RC and Giuliani F: Granzyme B-inhibitor serpina3n induces neuroprotection in vitro and in vivo. 12:1572015.

17 

Duan L, Zhang X, Panpan YU, Feng YU, Liu W and Song N: Effects of cholecystokinin-octopeptide on endoplasmic reticulum stress induced by LPS in mouse monocyte RAW264.7 cells. Journal of Shandong University. 53:16–20. 2015.

18 

Afonina IS, Cullen SP and Martin SJ: Cytotoxic and non-cytotoxic roles of the CTL/NK protease granzyme B. Immunol Rev. 235:105–116. 2010. View Article : Google Scholar : PubMed/NCBI

19 

Tak PP, Kummer JA, Hack CE, Daha MR, Smeets TJ, Erkelens GW, Meinders AE, Kluin PM and Breedveld FC: Granzyme-positive cytotoxic cells are specifically increased in early rheumatoid synovial tissue. Arthritis Rheum. 37:1735–1743. 1994. View Article : Google Scholar : PubMed/NCBI

20 

Goldbach-Mansky R, Suson S, Wesley R, Hack CE, El-Gabalawy HS and Tak PP: Raised granzyme B levels are associated with erosions in patients with early rheumatoid factor positive rheumatoid arthritis. Ann Rheum Dis. 64:715–721. 2005. View Article : Google Scholar : PubMed/NCBI

21 

Metkar SS, Menaa C, Pardo J, Wang B, Wallich R, Freudenberg M, Kim S, Raja SM, Shi L, Simon MM and Froelich CJ: Human and mouse granzyme A induce a proinflammatory cytokine response. Immunity. 29:720–733. 2008. View Article : Google Scholar : PubMed/NCBI

22 

Rossol M, Heine H, Meusch U, Quandt D, Klein C, Sweet MJ and Hauschildt S: LPS-induced cytokine production in human monocytes and macrophages. Crit Rev Immunol. 31:379–446. 2011. View Article : Google Scholar : PubMed/NCBI

23 

Ashbrook MJ, McDonough KL, Pituch JJ, Christopherson PL, Cornell TT, Selewski DT, Shanley TP and Blatt NB: Citrate modulates lipopolysaccharide-induced monocyte inflammatory responses. Clin Exp Immunol. 180:520–530. 2015. View Article : Google Scholar : PubMed/NCBI

24 

Kwon SH, Ma SX, Hong SI, Lee SY and Jang CG: Lonicera japonica THUNB. Extract Inhibits Lipopolysaccharide-Stimulated Inflammatory Responses by Suppressing NF-κB Signaling in BV-2 Microglial Cells. J Med Food. 18:762–775. 2015. View Article : Google Scholar : PubMed/NCBI

25 

Haile Y, Carmine-Simmen K, Olechowski C, Kerr B, Bleackley RC and Giuliani F: Granzyme B-inhibitor serpina3n induces neuroprotection in vitro and in vivo. J Neuroinflammation. 12:1572015. View Article : Google Scholar : PubMed/NCBI

26 

Liu H, Yin JJ, Cao MM, Liu GD, Su Y and Li YB: Endoplasmic reticulum stress induced by lipopolysaccharide is involved in the association between inflammation and autophagy in INS-1 cells. Mol Med Rep. 16:5787–5792. 2017. View Article : Google Scholar : PubMed/NCBI

27 

Prell T, Lautenschläger J, Weidemann L, Ruhmer J, Witte OW and Grosskreutz J: Endoplasmic reticulum stress is accompanied by activation of NF-κB in amyotrophic lateral sclerosis. J Neuroimmunol. 270:29–36. 2014. View Article : Google Scholar : PubMed/NCBI

28 

Liu H, Qian J, Wang F, Sun X, Xu X, Xu W and Zhang X and Zhang X: Expression of two endoplasmic reticulum stress markers, GRP78 and GADD153, in rat retinal detachment model and its implication. Eye (Lond). 24:137–144. 2010. View Article : Google Scholar : PubMed/NCBI

29 

Lee AS: The ER chaperone and signaling regulator GRP78/BiP as a monitor of endoplasmic reticulum stress. Methods. 35:373–381. 2005. View Article : Google Scholar : PubMed/NCBI

30 

Chen G, Li X, Huang M, Li M, Zhou X, Li Y and Bai J: Thioredoxin-1 increases survival in sepsis by inflammatory response through suppressing endoplasmic reticulum stress. Shock. 46:67–74. 2016. View Article : Google Scholar : PubMed/NCBI

31 

Lin WC, Chuang YC, Chang YS, Lai MD, Teng YN, Su IJ, Wang CC, Lee KH and Hung JH: Endoplasmic reticulum stress stimulates p53 expression through NF-κB activation. PLoS One. 7:e391202012. View Article : Google Scholar : PubMed/NCBI

32 

Ghosh S and Karin M: Missing pieces in the NF-kappaB puzzle. Cell. 109 Suppl:S81–S96. 2002. View Article : Google Scholar : PubMed/NCBI

33 

Willy JA, Young SK, Stevens JL, Masuoka HC and Wek RC: CHOP links endoplasmic reticulum stress to NF-κB activation in the pathogenesis of nonalcoholic steatohepatitis. Mol Biol Cell. 26:2190–2204. 2015. View Article : Google Scholar : PubMed/NCBI

34 

Legler DF, Micheau O, Doucey MA, Tschopp J and Bron C: Recruitment of TNF Receptor 1 to Lipid Rafts Is Essential for TNFalpha-Mediated NF-kappaB Activation. Immunity. 18:655–664. 2003. View Article : Google Scholar : PubMed/NCBI

35 

Palacios MM and Bleackley C: Characterization of Critical Residues of the Granzyme B Inhibitor, Serpina3n (50.43). J Immunol. 182:43–50. 2009.

36 

Han YP, Yan C and Garner WL: Proteolytic activation of matrix metalloproteinase-9 in skin wound healing is inhibited by alpha-1-Antichymotrypsin. J Invest Dermatol. 128:2334–2342. 2008. View Article : Google Scholar : PubMed/NCBI

Related Articles

  • Abstract
  • View
  • Download
  • Twitter
Copy and paste a formatted citation
Spandidos Publications style
Wang L, Jiang S, Xiao L, Chen L, Zhang Y and Tong J: Inhibition of granzyme B activity blocks inflammation induced by lipopolysaccharide through regulation of endoplasmic reticulum stress signaling in NK92 cells. Mol Med Rep 18: 580-586, 2018.
APA
Wang, L., Jiang, S., Xiao, L., Chen, L., Zhang, Y., & Tong, J. (2018). Inhibition of granzyme B activity blocks inflammation induced by lipopolysaccharide through regulation of endoplasmic reticulum stress signaling in NK92 cells. Molecular Medicine Reports, 18, 580-586. https://doi.org/10.3892/mmr.2018.8995
MLA
Wang, L., Jiang, S., Xiao, L., Chen, L., Zhang, Y., Tong, J."Inhibition of granzyme B activity blocks inflammation induced by lipopolysaccharide through regulation of endoplasmic reticulum stress signaling in NK92 cells". Molecular Medicine Reports 18.1 (2018): 580-586.
Chicago
Wang, L., Jiang, S., Xiao, L., Chen, L., Zhang, Y., Tong, J."Inhibition of granzyme B activity blocks inflammation induced by lipopolysaccharide through regulation of endoplasmic reticulum stress signaling in NK92 cells". Molecular Medicine Reports 18, no. 1 (2018): 580-586. https://doi.org/10.3892/mmr.2018.8995
Copy and paste a formatted citation
x
Spandidos Publications style
Wang L, Jiang S, Xiao L, Chen L, Zhang Y and Tong J: Inhibition of granzyme B activity blocks inflammation induced by lipopolysaccharide through regulation of endoplasmic reticulum stress signaling in NK92 cells. Mol Med Rep 18: 580-586, 2018.
APA
Wang, L., Jiang, S., Xiao, L., Chen, L., Zhang, Y., & Tong, J. (2018). Inhibition of granzyme B activity blocks inflammation induced by lipopolysaccharide through regulation of endoplasmic reticulum stress signaling in NK92 cells. Molecular Medicine Reports, 18, 580-586. https://doi.org/10.3892/mmr.2018.8995
MLA
Wang, L., Jiang, S., Xiao, L., Chen, L., Zhang, Y., Tong, J."Inhibition of granzyme B activity blocks inflammation induced by lipopolysaccharide through regulation of endoplasmic reticulum stress signaling in NK92 cells". Molecular Medicine Reports 18.1 (2018): 580-586.
Chicago
Wang, L., Jiang, S., Xiao, L., Chen, L., Zhang, Y., Tong, J."Inhibition of granzyme B activity blocks inflammation induced by lipopolysaccharide through regulation of endoplasmic reticulum stress signaling in NK92 cells". Molecular Medicine Reports 18, no. 1 (2018): 580-586. https://doi.org/10.3892/mmr.2018.8995
Follow us
  • Twitter
  • LinkedIn
  • Facebook
About
  • Spandidos Publications
  • Careers
  • Cookie Policy
  • Privacy Policy
How can we help?
  • Help
  • Live Chat
  • Contact
  • Email to our Support Team