Association of rs2249863 and rs1233334 polymorphisms of the upstream region of HLA-G gene with recurrent spontaneous abortion in women from Northwest of Iran during 2018-2019

Document Type : Original Article


1 M.Sc. in Genetics, School of Basic Sciences, Islamic Azad University, Tabriz, Iran.

2 Associate professor, Department of Molecular Genetic, Faculty of Natural Sciences, University of Tabriz, Tabriz. Iran.

3 M.Sc. in Genetics, School of Natural Sciences, University of Tabriz, Tabriz, Iran.


Introduction: Genetic polymorphisms are one of the possible causes of recurrent abortion with unknown cause. This study was performed with aim to investigate the association and frequency of rs2249863 and rs1233334 polymorphisms of the upstream region of HLA-G gene with the recurrent spontaneous abortion in women from Northwest of Iran.
Methods: This case-control study was performed on 160 women from Northwest of Iran referring to the Madar infertility clinic of Tabriz between 2018 and 2019. Among them, 80 women with a history of recurrent spontaneous abortion (at least 2 abortions) were considered as case group and 80 women with at least one child and without any history of miscarriage as control group. After collecting blood samples, genome DNA was extracted and the polymorphisms were evaluated by PCR and sequencing technique. Data were analyzed using SPSS software (version 22) and Fisher test. P <0.05 was considered statistically significant.    
Results: The frequencies of allele G and genotype GG in the rs2249863 polymorphism were significantly different between the case and control groups (P<0.05). In the rs1233334 polymorphism, although the frequencies of allele C and genotype CC were different between the case and control groups, but the differences were not statistically significant (P>0.05). Moreover, haplotype analysis identified the CG as the commonest haplotype that its frequency was significantly different between the case and control groups (P<0.05).
Conclusion: The rs2249863 polymorphism and CG haplotype of the rs2249863 and rs1233334 polymorphisms are associated with the recurrent spontaneous abortion in the studied population and they could be considered as potential risk factors for the disease.


  1. Ford HB, Schust DJ. Recurrent pregnancy loss: etiology, diagnosis, and therapy. Rev Obstet Gynecol 2009; 2(2):76-83.
  2. Christiansen OB, Nielsen HS, Kolte AM. Future directions of failed implantation and recurrent miscarriage research. Reprod Biomed Online 2006; 13(1):71-83.
  3. Pandey MK, Rani R, Agrawal S. An update in recurrent spontaneous abortion. Arch Gynecol Obstet 2005; 272(2):95-108.
  4. Castelli EC, Ramalho J, Porto IO, Lima TH, Felício LP, Sabbagh A, et al. Insights into HLA-G Genetics Provided by Worldwide Haplotype Diversity. Front Immunol 2014; 5:476.
  5. Castelli EC, Mendes-Junior CT, Deghaide NH, de Albuquerque RS, Muniz YC, Simões RT, et al. The genetic structure of 3'untranslated region of the HLA-G gene: polymorphisms and haplotypes. Genes Immun 2010; 11(2):134-41.
  6. Farzad F, Abediankenari S, Rahmani Z, Hashemi-Soteh MB, Hosseinikhah Z, Naghavian E. The role of HLA-G4 and G5 in threatened-abortion women. Journal of Mazandaran University of Medical Sciences 2013; 23(106):2-10.
  7. Ellis SA, Sargent IL, Redman CW, McMichael AJ. Evidence for a novel HLA antigen found on human extravillous trophoblast and a choriocarcinoma cell line. Immunology 1986; 59(4):595-601.
  8. Hviid TV. HLA-G in human reproduction: aspects of genetics, function and pregnancy complications. Hum Reprod Update 2006; 12(3):209-32.
  9. Moreau P, Contu L, Alba F, Lai S, Simoes R, Orrù S, et al. HLA-G gene polymorphism in human placentas: possible association of G*0106 allele with preeclampsia and miscarriage. Biol Reprod 2008; 79(3):459-67.
  10. González A, Rebmann V, LeMaoult J, Horn PA, Carosella ED, Alegre E. The immunosuppressive molecule HLA-G and its clinical implications. Crit Rev Clin Lab Sci 2012; 49(3):63-84.
  11. Shiroishi M, Kuroki K, Rasubala L, Tsumoto K, Kumagai I, Kurimoto E, et al. Structural basis for recognition of the nonclassical MHC molecule HLA-G by the leukocyte Ig-like receptor B2 (LILRB2/LIR2/ILT4/CD85d). Proc Natl Acad Sci U S A 2006; 103(44):16412-7.
  12. Hviid TV, Rizzo R, Melchiorri L, Stignani M, Baricordi OR. Polymorphism in the 5′ upstream regulatory and 3′ untranslated regions of the HLA-G gene in relation to soluble HLA-G and IL-10 expression. Human immunology 2006; 67(1-2):53-62.
  13. Ober C, Aldrich CL, Chervoneva I, Billstrand C, Rahimov F, Gray HL, et al. Variation in the HLA-G promoter region influences miscarriage rates. Am J Hum Genet 2003; 72(6):1425-35.
  14. Berger DS, Hogge WA, Barmada MM, Ferrell RE. Comprehensive analysis of HLA-G: implications for recurrent spontaneous abortion. Reprod Sci 2010; 17(4):331-8.
  15. Miller SA, Dykes DD, Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 1988; 16(3):1215.
  16. Christiansen OB, Nybo Andersen AM, Bosch E, Daya S, Delves PJ, Hviid TV, et al. Evidence-based investigations and treatments of recurrent pregnancy loss. Fertil Steril 2005; 83(4):821-39.
  17. Hunt JS, Jadhav L, Chu W, Geraghty DE, Ober C. Soluble HLA-G circulates in maternal blood during pregnancy. American journal of obstetrics and gynecology 2000; 183(3):682-8.
  18. Donadi EA, Castelli EC, Arnaiz-Villena A, Roger M, Rey D, Moreau P. Implications of the polymorphism of HLA-G on its function, regulation, evolution and disease association. Cell Mol Life Sci 2011; 68(3):369-95.
  19. Nowak I, Malinowski A, Barcz E, Wilczyński JR, Wagner M, Majorczyk E, et al. Possible Role of HLA-G, LILRB1 and KIR2DL4 Gene Polymorphisms in Spontaneous Miscarriage. Arch Immunol Ther Exp (Warsz) 2016; 64(6):505-514.
  20. Carlini F, Traore K, Cherouat N, Roubertoux P, Buhler S, Cortey M, et al. HLA-G UTR haplotype conservation in the Malian population: association with soluble HLA-G. PLoS One 2013; 8(12):e82517.
  21. Agrawal D, Prakash S, Misra MK, Phadke SR, Agrawal S. Implication of HLA-G 5' upstream regulatory region polymorphisms in idiopathic recurrent spontaneous abortions. Reprod Biomed Online 2015; 30(1):82-91.
  22. Eskandari F, Akbari MT, Zare S. Investigation the Association between Factor XIII Gene Polymorphism G103T with Recurrent Miscarriage in Iranian Patients. Iran J Obstet Gynecol Infertil 2013; 16(76):19-24.
  23. Farhangi H, Badiei Z. Report of some cases of recurrent pregnancy loss due to congenital deficiency of factor XIII. Iran J Obstet Gynecol Infertil 2015; 17(138):18-21.
  24. Khanbarari F, Vakili M, Samadi M. The relationship between rs2488457 Polymorphism of PTPN22 Gene and Recurrent Pregnancy Loss (RPL). Iran J Obstet Gynecol Infertil 2017; 20(9):49-55.
  25. Hajifathaliya Z, Najafipour R, Modarressi MH, Savad S, Rashvand Z. Association between the HLA-G* 0105N polymorphism and recurrent abortion in women. The Journal of Qazvin University of Medical Sciences 2018; 21(6):30-37.
  26. Alizadeh N, Mosaferi E, Farzadi L, Majidi J, Monfaredan A, Yousefi B, et al. Frequency of null allele of Human Leukocyte Antigen-G (HLA-G) locus in subjects to recurrent miscarriage. Int J Reprod Biomed 2016;4(7):459-64.
  27. Quintero-Ronderos P, Laissue P. Genetic Variants Contributing to Early Recurrent Pregnancy Loss Etiology Identified by Sequencing Approaches. Reprod Sci 2019: 1933719119831769.
  28. Quintero-Ronderos P, Mercier E, Fukuda M, González R, Suárez CF, Patarroyo MA, et al. Novel genes and mutations in patients affected by recurrent pregnancy loss. PLoS One 2017; 12(10):e0186149.
  29. Qiao Y, Wen J, Tang F, Martell S, Shomer N, Leung PC, et al. Whole exome sequencing in recurrent early pregnancy loss. Mol Hum Reprod 2016; 22(5):364-72.
  30. Craenmehr MHC, Nederlof I, Cao M, Drabbels JJM, Spruyt-Gerritse MJ, Anholts JDH, et al. Increased HLA-G Expression in Term Placenta of Women with a History of Recurrent Miscarriage Despite Their Genetic Predisposition to Decreased HLA-G Levels. Int J Mol Sci 2019; 20(3):625.
  31. ‏Dias FC, Bertol BC, Poras I, Souto BM, Mendes-Junior CT, Castelli EC, et al. The genetic diversity within the 1.4 kb HLA-G 5' upstream regulatory region moderately impacts on cellular microenvironment responses. Sci Rep 2018; 8(1):5652.
  32. Meuleman T, Drabbels J, van Lith JMM, Dekkers OM, Rozemuller E, Cretu-Stancu M, et al. Lower frequency of the HLA-G UTR-4 haplotype in women with unexplained recurrent miscarriage. J Reprod Immunol 2018; 126:46-52.