Anti-annexin A5 antibodies and 25-hydroxy-cholecalciferol in female patients with primary antiphospholipid syndrome

  • Mirjana Bećarević
  • Marija Sarić
  • Ljudmila Stojanovich
  • Duško Mirković
  • Violeta Dopsaj
  • Svetlana Ignjatović
Original Article


Vascular antiphospholipid syndrome (VAPS) and obstetric (OAPS) are different entities because some patients only develop thrombosis (without recurrent pregnancy losses) and vice versa. Only two articles have reported that low 25-hydroxy-cholecalciferol (vitamin D3, VD3) levels were not correlated with the presence of conventional antiphospholipid antibodies (aPL Abs: anticardiolipin (aCL), anti-beta2glycoprotein I (aβ2gpI), and lupus anticoagulant (LA)), but no article analyzed the association of VD3 and anti-annexin A5 (aanxA5) Abs. The aim of our study was to investigate the association between VD3, multiple positivity of conventional aPL and aanxA5 Abs levels only in female OAPS vs. VAPS. Our study included 62 consecutive female PAPS patients. Concentrations of Abs were measured by ELISA, while VD3 levels were determined by immunochemiluminescence. Only 10/62 (16.13%) had sufficient (≥ 30 ng/ml) VD3 levels, while 48/62 (77.42%) and 4/62 (6.45%) had insufficiency and VD3 deficiency, respectively. Statistically significant VD3 deficiency was noticed in VAPS (vs. OAPS, P = 0.013). A negative correlation between VD3 levels and the age of patients was noticed (r = − 0.493, P = 0.032) only in VAPS subgroup. Multiple positivity of aPL and aanxA5 Abs was not associated with VD3 deficiency. Newly emerging aPL Abs, such as aanxA5 Abs, or their combinations with classical aPL Abs are not associated with VD3 deficiency in neither OAPS nor VAPS patients. Due to its immunomodulatory roles in B-Ly homeostasis, supplementation with VD3 should be considered in APS, at least in subgroup with severe form of the disease, i.e., VAPS.


25-hydroxy-cholecalciferol Anti-annexin A5 antibodies Antiphospholipid antibodies Antiphospholipid syndrome Vitamin D3 



We would like to thank all patients who participated in this study and our colleagues from numerous clinics in Serbia.


The present work was supported by the Ministry of Science, Education and Technological Development of the Republic of Serbia on the basis of contract No. 175036. In addition, our work was supported by the research grant numbers 175041 and TR 32040 (for 2011–2018), issued by the Ministry of Science of the Republic of Serbia.

The funding source had no involvement in study design; in the collection, analysis and interpretation of data; in the writing of the report; and in the decision to submit this article for publication.

Compliance with ethical standards

Ethical standards

All procedures performed in our study were in accordance with Helsinki declaration (and its later amendments) and with the ethical standards of the institutional ethical committee. Informed consent was obtained from all individual participants included in the study.



Informed consent

Informed consent was obtained from all individual participants included in the study.


None of the authors has any potential financial conflict of interest related to this manuscript. The authors alone are responsible for the content and writing of the paper.


  1. 1.
    Miyakis S, Lockshin MD, Atsumi T et al (2006) International consensus statement on an update of the classification criteria for definite antiphospholipid syndrome. J Thromb Haemost 4:295–306CrossRefPubMedGoogle Scholar
  2. 2.
    Bertolaccini ML, Ammengual O, Andreolii L et al (2014) 14th international congress on antiphospholipid antibodies task force. Report on antiphospholipid syndrome laboratory diagnostics and trends. Autoimmun Rev 13:917–930CrossRefPubMedGoogle Scholar
  3. 3.
    Bećarević M (2017) Antibodies against complement components: relevance for the antiphospholipid syndrome-biomarkers of the disease and biopharmaceuticals. Curr Rheumatol Rep 19(40):1–9. CrossRefGoogle Scholar
  4. 4.
    Bećarević M (2017) Detrimental roles of TNF-alpha in the antiphospholipid syndrome and de novo synthesis of antiphospholipid antibodies induced by biopharmaceuticals against TNF-alpha. J Thromb Thrombolysis 44:565–570CrossRefPubMedGoogle Scholar
  5. 5.
    Bećarević M (2016) The IgG and IgM isotypes of anti-annexin A5 antibodies: relevance for primary antiphospholipid syndrome. J Thromb Thrombolysis 42(4):552–557. CrossRefPubMedGoogle Scholar
  6. 6.
    Bećarević M, Ignjatović S (2016) Proinflammatory proteins in female and male patients with primary antiphospholipid syndrome: preliminary data. Clin Rheumatol 35(10):2477–2483. CrossRefPubMedGoogle Scholar
  7. 7.
    Bećarević M (2016) TNF-alpha and annexin A2: inflammation in thrombotic primary antiphospholipid syndrome. Rheumatol Int 36(12):1649–1656. CrossRefPubMedGoogle Scholar
  8. 8.
    Bećarević M, Mirković D, Ignjatović S (2016) Double positivity of the IgG isotype of both anticardiolipin and anti-β2gpI antibodies is associated with the highest number of vascular impairment parameters in patients with primary antiphospholipid syndrome: preliminary data. Clin Rheumatol 35(12):2947–2954. CrossRefPubMedGoogle Scholar
  9. 9.
    D’Ippolito S, Meroni PL, Koike T, Veglia M, Scambia G, Di Simone N (2014) Obstetric antiphospholipid syndrome: a recent classification for an old defined disorder. Autoimmun Rev 13:901–908CrossRefPubMedGoogle Scholar
  10. 10.
    Stojanovich L, Markovic O, Marisavljevic D, Elezovic I, Ilijevski N, Stanisavljevic N (2011) Influence of antiphospholipid antibody levels and type on thrombotic manifestations: results from the Serbian National Cohort Study. Lupus 21:1–8. CrossRefGoogle Scholar
  11. 11.
    Stojanovich L, Kontic M, Djokovic A, Marisavljevic D, Ilijevski N, Stanisavljevic N, Mikovic Z, Petkovic M, Kovcin V (2013) Association between systemic non-criteria APS manifestations and antibody type and level: results from the Serbian National Cohort Study. Clin Exp Rheumatol 31(5):756–760PubMedGoogle Scholar
  12. 12.
    Vasile M, Corinaldesi C, Antinozzi C, Crescioli C (2017) Vitamin D in autoimmune rheumatic diseases: a view inside gender differences. Pharmacol Res 117:228–241CrossRefPubMedGoogle Scholar
  13. 13.
    Heine GK, Anton B, Henz M, Worm M (2002) 1,25-dihydroxyvitamin D3 inhibits anti-CD40 plus IL-4-mediated IgE production in vitro. Eur J Immunol 32:3395–3404PubMedGoogle Scholar
  14. 14.
    Muller K, Heilmann C, Poulsen LK, Barington T, Bendtzen K (1991) The role of monocytes and T-cells in 1,25-dihydroxyvitamin-D3 mediated inhibition of B-cell function-in vitro. Immunopharmacology 21:121–128CrossRefPubMedGoogle Scholar
  15. 15.
    Chen S, Sims GP, Chen XX, Gu YY, Chen S, Lipsky PE (2007) Modulatory effects of 1,25-Dihydroxyvitamin D3 on human B cell differentiation. J Immunol 179:1634–1647. CrossRefPubMedGoogle Scholar
  16. 16.
    Agmon-Levin N, Blank M, Zandman-Goddard Z et al (2011) Vitamin D: an instrumental factor in the anti-phospholipid syndrome by inhibition of tissue factor expression. Ann Rheum Dis 70:145–150. CrossRefPubMedGoogle Scholar
  17. 17.
    Paupitz JA, Freire de Carvalho J, Caparbo VF, Klack K, Pereira RMR (2010) Primary antiphospholipid syndrome in premenopausal women: low vitamin D, high fat mass and maintained bone mineral mass. Lupus 19:1302–1306CrossRefPubMedGoogle Scholar
  18. 18.
    Bećarević M, Stojanovich L, Ignjatović S, Dopsaj V (2016) The IgM isotype of anti-annexin A5 antibodies and multiple positivity of conventional antiphospholipid antibodies: increasing the number of clinical manifestations of primary antiphospholipid syndrome. Clin Rheumatol 35(5):1361–1365. issn: 0770-3198CrossRefPubMedGoogle Scholar
  19. 19.
    Brandt JT, Triplett DA, Alving B, Scharrer J (1995) Criteria for the diagnosis of lupus anticoagulants: an update. On behalf of the Subcommittee on Lupus Anticoagulant/Antiphospholipid Antibody of the Scientific and Standardisation Committee of the International Society of Thrombosis and Haemostasis. Thromb Haemost 74(4):1185–1190PubMedCrossRefGoogle Scholar
  20. 20.
    Ross AC, Manson JE, Abrams SA, Aloia JF, Brannon PM, Clinton SK et al (2011) The 2011 report on dietary reference intakes for calcium and vitamin D from the Institute of Medicine: what clinicians need to know. J Clin Endocrinol Metab 96(1):53–58CrossRefPubMedGoogle Scholar
  21. 21.
    Jelić S, Nikolić D, Marisavljević D, Stojanovich L (2017) Pathogenic and therapeutic role of vitamin D in antiphospholipid syndrome patients. In: A critical evaluation of vitamin D-clinical overview. Intech, pp 180–204.
  22. 22.
    Klack K, Freire de Carvalho J (2010) High frequency of vitamin D insufficiency in primary antiphospolipid syndrome. Joint Bone Spine 77:482–491CrossRefGoogle Scholar
  23. 23.
    Andreoli L, Piantoni S, Dall’Ara F, Allegri F, Meroni PL, Tincani A (2012) Vitamin D and antiphospholipid syndrome. Lupus 21:736–740CrossRefPubMedGoogle Scholar
  24. 24.
    Ota K, Dambaeva S, Han AR, Beaman K, Gilman-Sachs A, Kwak-Kim J (2014) Vitamin D deficiency may be a risk factor for recurrent pregnancy losses by increasing cellular immunity and autoimmunity. Hum Reprod 29:208–219CrossRefPubMedGoogle Scholar
  25. 25.
    Ota K, Dambaeva S, Kim MW, Han AR, Fukui A, Gilman-Sachs A et al (2015) 1,25-Dihydroxyvitamin D3 regulates NK-cell cytotoxicity, cytokine secretion, and degranulation in women with recurrent pregnancy losses. Eur J Immunol 45:3188–3199CrossRefPubMedGoogle Scholar
  26. 26.
    Kwak-Kima J, Skariah A, Wua L, Salazar D, Sung N, Ota K (2016) Humoral and cellular autoimmunity in women with recurrent pregnancy losses and repeated implantation failures: a possible role of vitamin D. Autoimmun Rev 15:943–947CrossRefGoogle Scholar
  27. 27.
    Arnold J, Holmes Z, Pickering W, Farmer C, Regan L, Cohen H (2001) Anti-beta2glicoprotein I and of anti-annexin V antibodies in women with recurrent miscarriages. Br J Haematol 113:911–914CrossRefPubMedGoogle Scholar
  28. 28.
    Bećarević M, Singh S, Majkić-Singh N (2008) Oxidized LDL, anti-oxidized LDL and anti-annexin A5 antibodies in primary antiphospholipid syndrome. Clin Lab 54:97–101PubMedGoogle Scholar
  29. 29.
    Rand JH, Arslan AA, Wu XX, Wein R, Mulholland J, Shah M, van Heerde WL, Reutelingsperger CP, Lockwood CJ, Kuczynski E (2006) Reduction of circulating annexin A5 levels and resistance to annexin A5 anticoagulant activity in women with recurrent spontaneous pregnancy losses. Am J Obstet Gynecol 194:182–188CrossRefPubMedGoogle Scholar
  30. 30.
    Trouve P, Kerbiriou M, le Hir S, Benz N, Férec C (2012) Surface plasmon resonance shows a gender difference in circulating annexin A5 in human. Talanta 93:219–223CrossRefPubMedGoogle Scholar

Copyright information

© International League of Associations for Rheumatology (ILAR) 2018

Authors and Affiliations

  1. 1.University of Novi SadFaculty of Medicine, Department of PharmacyNovi SadSerbia
  2. 2.University of BelgradeFaculty of Pharmacy, Department for Medical BiochemistryBelgradeSerbia
  3. 3.University of BelgradeUniversity Medical Center “Bežanijska Kosa”BelgradeSerbia

Personalised recommendations