Expression of NKG2D Receptors in CD3+ and CD56+ Cells in Blood Samples of Pre-eclampsia Patients and the Effect of Silymarin on Their Activity in Vitro

Authors

1 Professor, Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

2 Associate Professor, Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

3 Associate Professor, Department of Obstetrics and Gynecology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

4 MSc Student, Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

5 MSc, Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

Abstract

Background: Increased NK cells or their cytokine activity in endometrium and peripheral blood of mothers with pre-eclampsia have been reported. NKG2D is a cell surface receptor that can be used to detect NK cell and lymphocytic activity. Plant flavonoids such as silymarin has immunomodulatory effects and potentially inhibit the disease progression. Therefore, an in vitro experiment was designed to investigate the immunomodulatory effects of silymarin in pre-eclampsia.
Methods: Blood samples from preeclamptic and pregnant and non-pregnant women were analyzed by Isolation of peripheral blood mononuclear cells (PBMCs) using Ficoll-Paque. Flow cytometry was used for marker assay including NKG2D expression on CD56+CD3- and CD3+ cells. Also, different concentrations of silymarin were applied, andNKG2D expression on the cells were considered, and IFNγ production in the cell culture medium were evaluated using ELISA.
Findings: CD3-CD56+ cells and NKG2D receptor expression were augmented in PBMCs in the pre-eclampsia group (P = 0.03). However, treatment of PBMCs with silymarin solutions decrease NKG2D markers on the lymphocytes. PBMCs treated cells by silymarin in culture medium indicates reduce IFNɣ production even in the presence of IL-2 stimulation by 2-3 folds (P = 0.0001).
Conclusion: Increase in NK cell population, NKG2D receptor expression and IFNɣ production play a crucial role in the samples of preeclamptic patients, and well express involvement of immunological factors of the disease. The effectiveness of silymarin in cell culture medium and modulation of inflammatory factors necessitate the design of clinical model for using silymarin in patients by risk of increased NK activity in pre-eclampsia patients.

Keywords

References

  1. Rambaldi MP, Weiner E, Mecacci F, Bar J, Petraglia F. Immunomodulation and preeclampsia. Best Pract Res Clin Obstet Gynaecol 2019; 60: 87-96.
  2. Andalib A, Rezaie A, Oreizy F, Shafiei K, Baluchi S. A study on stress, depression and NK cytotoxic potential in women with recurrent spontaneous abortion. Iran J Allergy Asthma Immunol 2006; 5(1): 9-16.
  3. Granger JP, LaMarca BB, Cockrell K, Sedeek M, Balzi C, Chandler D, et al. Reduced uterine perfusion pressure (RUPP) model for studying cardiovascular-renal dysfunction in response to placental ischemia. Methods Mol Med 2006; 122: 383-92.
  4. Steegers EA, von Dadelszen P, Duvekot JJ, Pijnenborg R. Pre-eclampsia. Lancet 2010; 376(9741): 631-44.
  5. Kobayashi H, Ichikawa M, Akasaka J, Tsunemi T, Sado T. Immune-related pathophysiological causes relevant to a subset of patients with preeclampsia (Review). World Acad Sci J 2019; 1(2): 59-66.
  6. Park DW, Lee HJ, Park CW, Hong SR, Kwak-Kim J, Yang KM. Peripheral blood NK cells reflect changes in decidual NK cells in women with recurrent miscarriages. Am J Reprod Immunol 2010; 63(2): 173-80.
  7. Fukui A, Funamizu A, Fukuhara R, Shibahara H. Expression of natural cytotoxicity receptors and cytokine production on endometrial natural killer cells in women with recurrent pregnancy loss or implantation failure, and the expression of natural cytotoxicity receptors on peripheral blood natural killer cells in pregnant women with a history of recurrent pregnancy loss. J Obstet Gynaecol Res 2017; 43(11): 1678-86.
  8. Costello RT, Fauriat C, Sivori S, Marcenaro E, Olive D. NK cells: innate immunity against hematological malignancies? Trends Immunol 2004; 25(6): 328-33.
  9. Mincheva-Nilsson L, Nagaeva O, Chen T, Stendahl
    U, Antsiferova J, Mogren I, et al. Placenta-derived soluble MHC class I chain-related molecules down-regulate NKG2D receptor on peripheral blood mononuclear cells during human pregnancy: a possible novel immune escape mechanism for fetal survival. J Immunol 2006; 176(6): 3585-92.
  10. Bauer S, Groh V, Wu J, Steinle A, Phillips JH, Lanier LL, et al. Pillars Article: Activation of NK Cells and T Cells by NKG2D, a Receptor for Stress-Inducible MICA. Science. 1999. 285: 727-729. J Immunol 2018; 200(7): 2231-3.
  11. Stern-Ginossar N, Mandelboim O. An integrated view of the regulation of NKG2D ligands. Immunology 2009; 128(1): 1-6.
  12. Hedlund M, Stenqvist AC, Nagaeva O, Kjellberg L, Wulff M, Baranov V, et al. Human placenta expresses and secretes NKG2D ligands via exosomes that down-modulate the cognate receptor expression: evidence for immunosuppressive function. J Immunol 2009; 183(1): 340-51.
  13. Liu M, Zhen X, Song H, Chen J, Sun X, Li X, et al. Low-dose lymphocyte immunotherapy rebalances the peripheral blood Th1/Th2/Treg paradigm in patients with unexplained recurrent miscarriage. Reprod Biol Endocrinol 2017; 15(1): 95.
  14. Bachmayer N, Sohlberg E, Sundström Y, Hamad RR, Berg L, Bremme K, et al. Women with pre-eclampsia have an altered NKG2A and NKG2C receptor expression on peripheral blood natural killer cells. Am J Reprod Immunol 2009; 62(3): 147-57.
  15. Esmaeil N, Anaraki SB, Gharagozloo M, Moayedi B. Silymarin impacts on immune system as an immunomodulator: One key for many locks. Int Immunopharmacol 2017; 50: 194-201.
  16. Cristofalo R, Bannwart-Castro CF, Magalhães CG, Borges VT, Peraçoli JC, Witkin SS, et al. Silibinin attenuates oxidative metabolism and cytokine production by monocytes from preeclamptic women.
    Free Radic Res 2013; 47(4): 268-75.
  17. Gazák R, Walterová D, Kren V. Silybin and silymarin--new and emerging applications in medicine. Curr Med Chem 2007; 14(3): 315-38.
  18. Gabbe S, Nieby JR, Simpson JL. Obstetrics: Normal and Problem Pregnancies. Arch Fam Med 1998; 7(4): 386.
  19. von Dadelszen P, Menzies J, Magee LA. The complications of hypertension in pregnancy. Minerva Med 2005; 96(4): 287-302.
  20. Hidaka Y, Amino N, Iwatani Y, Kaneda T, Mitsuda N, Morimoto Y, et al. Changes in natural killer cell activity in normal pregnant and postpartum women: increases in the first trimester and postpartum period and decrease in late pregnancy. J Reprod Immunol 1991; 20(1): 73-83.
  21. Gharagozloo M, Javid EN, Rezaei A, Mousavizadeh K. Silymarin inhibits cell cycle progression and mTOR activity in activated human T cells: therapeutic implications for autoimmune diseases. Basic Clin Pharmacol Toxicol 2013; 112(4): 251-6.
  22. Wallace AE, Whitley GS, Thilaganathan B, Cartwright JE. Decidual natural killer cell receptor expression is altered in pregnancies with impaired vascular remodeling and a higher risk of pre-eclampsia. J Leukoc Biol 2015; 97(1): 79-86.
  23. Morishima C, Shuhart MC, Wang CC, Paschal DM, Apodaca MC, Liu Y, et al. Silymarin inhibits in vitro T-cell proliferation and cytokine production in hepatitis C virus infection. Gastroenterology 2010; 138(2): 671-81, 681.e1-2.
  24. Gharagozloo M, Jafari S, Esmaeil N, Javid EN, Bagherpour B, Rezaei A. Immunosuppressive effect of silymarin on mitogen-activated protein kinase signalling pathway: the impact on T cell proliferation and cytokine production. Basic Clin Pharmacol Toxicol 2013; 113(3): 209-14.
  25. Navabi F, Shaygannejad V, Abbasirad F, Vaez E, Hosseininasab F, Kazemi M, et al. Immunoregulatory effects of silymarin on proliferation and activation of Th1 cells isolated from newly diagnosed and IFN-ß(1b)-treated MS patients. Inflammation 2019; 42(1): 54-63.
Journal of Isfahan Medical School
Volume 40, Issue 661
1st Week, May
May and June 2022
Pages 95-102

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History

  • Receive Date: 02 May 2022
  • Accept Date: 02 May 2022

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