Molecular Biology Reports

, Volume 46, Issue 1, pp 83–95 | Cite as

Reciprocal regulation of pro-inflammatory Annexin A2 and anti-inflammatory Annexin A1 in the pathogenesis of rheumatoid arthritis

  • Vikram Haridas
  • Praveenkumar ShettyEmail author
  • E. Sarathkumar
  • Anil Bargale
  • Jamboor K. Vishwanatha
  • Vidya Patil
  • U. S. Dinesh
Original Article


Annexin A2 has been implicated in several immune modulated diseases including Rheumatoid arthritis (RA) pannus formation. The most relied treatment option for RA pathogenesis is glucocorticoids. Glucocorticoids regulate the synthesis, phosphorylation and cellular deposition of Annexin A1. This annexin mediates the anti-inflammatory actions of glucocorticoids. These two first characterized members of annexin superfamily proteins acts reciprocally, one as an anti-inflammatory and the other proinflammatory in nature. The possibility of these molecules as soluble biomarkers and as an upstream regulator of major cytokine devastation at RA microenvironment has not been previously explored. Current study elucidates the reciprocal regulation of these two annexins in RA pathogenesis. These Annexin A2/A1 and downstream cytokines in RA serum were analysed by ELISA. Western blot, Immunocytochemistry, immunoprecipitation and Immunohistochemistry were adapted to analyse these molecules in tissue and synovial fibroblasts and also in different experimental conditions. Significant increase in the level of Annexin A2 was noticed in naïve RA patients compared to controls (14.582 ± 1.766 ng/ml vs. 7.37 ± 1.450 ng/ml; p ≤ 0.001). In remission cases significant low levels was detected. On the contrary, significant decrease in the level of Annexin A1 was noticed in naïve RA patients compared to healthy controls (12.322 ± 2.91 vs. 16.998 ± 4.298 ng/ml; p ≤ 0.001), wherein remission cases serum Annexin A1 was significantly high. The knockdown of proinflammatory Annexin A2 by siRNA/antibody treatment could mimic the glucocorticoid treatment as which induced cellular Annexin A1 and membrane translocation resulting in the terminal action. Current data elucidating the regulatory interplay between Annexin A2 and Annexin A1 in RA pathogenesis.


Rheumatoid arthritis Proinflammatory mediators Annexin A2 Annexin A1 Inflammatory cytokines 



Authors would like to acknowledge the technical help rendered by Leonard Clinton D’souza and Dr. Vishwas Kaveeshwara.

Author contributions

Conceived the idea and designed the experiments: VH and PKS. Helped in clinical sample collection and correlated the clinical relevance to the study: VH, JKV and USD. Performed the experiments: SE, AB and PKS. Analysed the data (pathology): USD. Analysed the data (statistical analysis): PKS and AB. Wrote the paper: VH, JKV and PKS. Supervised the overall study: PKS.


This work was supported by funding from Arthritis Super speciality center Hubli, Karnataka. India.

Compliance with ethical standards

Conflict of interest

The authors have declared that no competing interests exist.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. The current study protocol was approved by SDM College of Medical Sciences and Hospital, Institutional Ethical Board (Approval No. 0507, 2015–2016).

Informed consent

Even though spare samples were used for the study, informed consent was obtained from all individual participants included in the study.


  1. 1.
    Firestein GS (2003) Evolving concepts of rheumatoid arthritis. Nature 423:356–361CrossRefPubMedGoogle Scholar
  2. 2.
    McInnes IB, Schett G (2007) Cytokines in the pathogenesis of rheumatoid arthritis. Nat Rev Immunol 7(6):429–442CrossRefPubMedGoogle Scholar
  3. 3.
    Albani S, Carson DA (1997) Etiology and pathogenesis of rheumatoid arthritis. In: Koopman WJ (ed) Arthritis and allied conditions. Lippincott Williams & Wilkins, Baltimore, p 979Google Scholar
  4. 4.
    Chomarat P, Kjeldsen-Kragh J, Quayle AJ, Natvig JB, Miossec P (1994) Different cytokine production profiles of γδ T cell clones: relation to inflammatory arthritis. Eur J Immunol 24(9):2087–2091CrossRefPubMedGoogle Scholar
  5. 5.
    Grassi W, De Angelis R, Lamanna G, Cervini C (1998) The clinical features of rheumatoid arthritis. Eur J Radiol 27(1):S18–S24CrossRefPubMedGoogle Scholar
  6. 6.
    Wang JG, Xu WD, Zhai WT, Li Y, Hu JW, Hu B,et al (2012) Disorders in angiogenesis and redox pathways are main factors contributing to the progression of rheumatoid arthritis. Arthritis Rheum 64(4):993–1004CrossRefPubMedGoogle Scholar
  7. 7.
    Bharadwaj A, Bydoun M, Holloway R, Waisman D (2013) Annexin A2 heterotetramer: structure and function. Int J MolSci 14(3):6259–6305CrossRefGoogle Scholar
  8. 8.
    Iaccarino L, Ghirardello A, Canova M, Zen M, Bettio S, Nalotto L et al (2011) Anti-annexins autoantibodies: their role as biomarkers of autoimmune diseases. Autoimmun Rev 10(9):553–558CrossRefPubMedGoogle Scholar
  9. 9.
    Li Q, Laumonnier Y, Syrovets T, Simmet T (2007) Plasmin triggers cytokine induction in human monocyte-derived macrophages. Arterioscler Thromb Vasc Biol 27(6):1383–1389CrossRefPubMedGoogle Scholar
  10. 10.
    Esposito I, Penzel R, Chaib-Harrireche M, Barcena U, Bergmann F, Riedl S et al (2006) Tenascin C and annexin II expression in the process of pancreatic carcinogenesis. J Pathol 208(5):673–685CrossRefPubMedGoogle Scholar
  11. 11.
    Genetos DC, Wong A, Watari S, Yellowley CE (2010) Hypoxia increases annexin A2 expression in osteoblastic cells via VEGF and ERK. Bone 47(6):1013–1019CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Yi J, Zhu Y, Jia Y, Jiang H, Zheng X, Liu D et al (2016) The annexin A2 promotes development in arthritis through neovascularization by amplification of hedgehog pathway. PLoS ONE 11(3):e0150363CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Annalisa P, Elise ED, Jameson TC, Sheila A, Klemen S, Catherine EC et al (2015) Annexin A2 is a target of autoimmune T and B cell responses associated with synovial fibroblast proliferation in patients with antibiotic-refractory lyme arthritis. Clin Immunol 160(2):336–341CrossRefGoogle Scholar
  14. 14.
    Peng C, Hai Y, Yaping T, Yiping X, Lili S, Ran B et al (2015) Annexin A2 as a target endothelial cell membrane autoantigen in Behçet’s disease. Sci Rep 5:8162CrossRefGoogle Scholar
  15. 15.
    Cesarman-Maus G, Rios-Luna NP, Deora AB, Huang B, Villa R, delCravioto MC et al (2006) Autoantibodies against the fibrinolytic receptor, annexin 2, in antiphospholipid syndrome. Blood 107:4375–4382CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Schuliga MYX, Langenbach S, Harris T, Stewart AG (2014) Extracellular annexin A2 mediates lung fibroblast cytokine production and proliferation: a potential role in pulmonary fibrosis. Am J Respir Crit Care Med 189:A2031Google Scholar
  17. 17.
    Zhang L, Peng X, Zhang Z, Feng Y, Jia X, Shi Y et al (2010) Subcellular proteome analysis unraveled annexin A2 related to immune liver fibrosis. J Cell Biochem 110:219–228PubMedGoogle Scholar
  18. 18.
    Zhao W, Zhang C, Shi M, Zhang J, Li M, Xue X et al (2014) The discoidin domain receptor 2/annexin A2/matrix metalloproteinase 13 loop promotes joint destruction in arthritis through promoting migration and invasion of fibroblast-likesynoviocytes. Arthritis Rheum 66:2355–2367CrossRefGoogle Scholar
  19. 19.
    Hetal BP, Kristin NK, Andre ÕLFS, Fulvio DÕA, Michael PS, Ana PG et al (2012) The impact of endogenous annexin A1 on glucocorticoid control of inflammatory arthritis. Ann Rheum Dis 71:1872–1880CrossRefGoogle Scholar
  20. 20.
    Ana PG, Kallyne KOM, Carine CD, Simone MB, Egle S, Sandra HPF et al (2013) Anti-inflammatory mechanisms of the annexin A1 protein and its mimetic peptide Ac2-26 in models of ocular inflammation in vivo and in vitro. J Immunol 190:5689–5701CrossRefGoogle Scholar
  21. 21.
    Yuan j, Eric FM, Wuqi S, Qiang C, Alastair S, Yuan HY (2013) Regulation of lung fibroblast activation by annexin A1. J Cell Physiol 228:476–484CrossRefGoogle Scholar
  22. 22.
    Annaleise VS, Paul H, Eric FM (2000) Annexin I and dexamethasone effects on phospholipase and cyclooxygenase activity in human synoviocytes. Mediators Inflamm 9:125–132CrossRefGoogle Scholar
  23. 23.
    Yuanhang Y, Paul H, Eric FM (1999) Inhibitory effect of annexin 1 on synovial inflammation in rat adjuvant arthritis. Arthritis Rheum 42(7):1538–1544CrossRefGoogle Scholar
  24. 24.
    Yang Y, Jia Y, Song W, Ngo D, Li Z, Morand EF (2009) Annexin 1 regulates inflammation and glucocorticoid sensitivity of human RA synovial fibroblasts. Arthritis Rheum 60(10):40Google Scholar
  25. 25.
    Shetty PK, Thamake SI, Biswas S et al (2012) Reciprocal regulation of annexin A2 and EGFR with Her-2 in Her-2 negative and herceptin-resistant breast cancer. PLoS ONE 7(9):e44299CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Valapala M, Vishwanatha JK (2011) Lipid raft endocytosis and exosomal transport facilitate extracellular trafficking of annexin A2. J Biol Chem 286:30911–30925CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Shetty PK, Anil B, Basavraj RP, Rajashekar M, Dinesh US, Jamboor KV et al (2016) Cell surface interaction of annexin A2 and galectin-3 modulates epidermal growth factor receptor signaling in Her-2 negative breast cancer cells. Mol Cell Biochem 411(1):221–233CrossRefPubMedGoogle Scholar
  28. 28.
    Iain BM, Georg S (2011) The pathogenesis of rheumatoid arthritis. N Engl J Med 365:2205–2219CrossRefGoogle Scholar
  29. 29.
    Solomon S, Kassahn D, Illges H (2005) The role of the complement and the Fcγ R system in the pathogenesis of arthritis. Arthritis Res Ther 7(4):129–135CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Swisher JF, Khatri U, Feldman GM (2007) Annexin A2 is a soluble mediator of macrophage activation. J Leukoc 82:1174–1184CrossRefGoogle Scholar
  31. 31.
    Iaccarino L et al (2011) Anti-annexins autoantibodies: their role as biomarkers of autoimmune diseases. Autoimmun Rev 10:553–558CrossRefPubMedGoogle Scholar
  32. 32.
    Mor A, Abramson SB, Pillinger MH (2005) The fibroblast-like synovial cell in rheumatoid arthritis: a key player in inflammation and joint destruction. Clin Immunol 115:118–128CrossRefPubMedGoogle Scholar
  33. 33.
    Yoshizawa T, Hammaker D, Boyle DL, Corr M, Flavell R, Davis R et al (2009) Role of MAPK kinase 6 in arthritis: distinct mechanism of action in inflammation and cytokine expression. J Immunol 183:1360–1367CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Ahmad MK, Roderick JF, Mauro P (2005) An overview of the effects of annexin 1 on cells involved in the inflammatory process. Mem Inst Oswaldo CruzRio de Janeiro 100(I):39–48Google Scholar
  35. 35.
    Kao W, Gu R, Jia Y, Xuemin W, Fan H, Harris J et al (2014) A formyl peptide receptor agonist suppresses inflammation and bone damage in arthritis. Br J Pharmacol 171:4087–4096CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Croxtall JD, Waheed S, Choudhury Q, Anand R, Flower RJ (1993) N-terminal peptide fragments of lipocortin-1 inhibit A549 cell growth and block EGF-induced stimulation of proliferation. Int J Cancer 54:153–158CrossRefPubMedGoogle Scholar
  37. 37.
    Hannon R, Croxtall JD, Getting SJ, Roviezzo F, Yona S, Paul-Clark MJ et al (2003) Aberrant inflammation and resistance to glucocorticoids in annexin 1−/− mouse. FASEB J 17:253–255CrossRefPubMedGoogle Scholar
  38. 38.
    Pepinsky RB, Sinclair LK (1986) Epidermal growth factor dependent phosphorylation of lipocortin. Nature 321(6065):81–84CrossRefPubMedGoogle Scholar
  39. 39.
    Ferlazzo V, D’agostino P, Milano S, Caruso R, Feo S, Cillari E et al (2003) Anti-inflammatory effects of annexin-1: stimulation of IL-10 release and inhibition of nitric oxide synthesis. IntImmunopharmacology 3:1363–1369CrossRefGoogle Scholar
  40. 40.
    Simon Y, Sigrid EM, Heinsbroek LP, Siamon G, Mauro P, Roderick JF (2006) Impaired phagocytic mechanism in annexin 1 null macrophages. Br J Pharmacol 148:469–477Google Scholar
  41. 41.
    Jesmond D, Lucy VN, Derek R, Dianne C, Kit-Yi L, Manno P (2008) Annexin 1 mediates the rapid anti-inflammatory effects of neutrophil derived microparticles. Blood 112:2512–2519CrossRefGoogle Scholar
  42. 42.
    Goulet F, Moore KG, Sartorelli AC (1992) Glycosylation of annexinI and annexin II. Biochem Biophys Res Commun 188(2):554–558CrossRefPubMedGoogle Scholar
  43. 43.
    Yangguang L, Ti W, Minzhi Z, Lixin L, Jun W, Xiaoli W et al (2013) Glycoproteomic analysis of tissues from patients with colon cancer using lectin microarrays and nano LC-MS/MS. Mol BioSyst 9:1877CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Arthritis Superspeciality CenterHubliIndia
  2. 2.Department of MedicineSDM College of Medical Sciences and HospitalDharwadIndia
  3. 3.Central Research LabSDM College of Medical Sciences and HospitalDharwadIndia
  4. 4.Department of BiochemistrySDM College of Medical Sciences and HospitalDharwadIndia
  5. 5.Department of Microbiology, Immunology and GeneticsUniversity of North Texas Health Science CenterFort WorthUSA
  6. 6.Department of PathologySDM College of Medical Sciences and HospitalDharwadIndia

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