Original Article

Association of HPA Antigens with Immune Thrombocytopenia: A Case-Control Study by PCR-SSP Method

Hematologic Cut-Off Assessment of Alpha-Thalassemia

Abstract

Background: Human platelet antigens (HPAs) play a clinically significant role in alloimmunization and the development of immune-mediated disorders such as immune thrombocytopenia (ITP), fetal and neonatal alloimmune thrombocytopenia (FNAIT), and post-transfusion purpura (PTP). Understanding the genetic profiles of HPAs is critical for preventing and treating these conditions. Given the limitations of serological methods in determining HPA genotypes, this study aims to investigate the association between the genotypes of HPA1, HPA2, HPA3, HPA4, and HPA15 antigens and autoimmune thrombocytopenia in Lorestan Province, utilizing the PCR-SSP method.

Materials and Methods: This case-control study involved 80 individuals diagnosed with ITP and 120 healthy controls. DNA samples were extracted using a commercial DNA extraction kit, with concentrations quantified via a Nanodrop spectrophotometer. Genotyping was performed using the PCR-SSP method with specific primers for each HPA gene. The genotype data were verified using previously established sample sets. The frequencies of each HPA genotype were recorded, and a comparative analysis was conducted between the patient and control groups to evaluate the study hypothesis.

Results: The results revealed that individuals carrying the HPA2b allele had a 5.31-fold increased risk of developing ITP, a statistically significant finding (P < 0.05, OR = 5.31). Similarly, the presence of the HPA15b allele was associated with a 6.54-fold increased risk (P < 0.05, OR = 6.54).

Conclusion: These findings, in conjunction with previous studies, suggest the need for larger-scale investigations across different populations. Such research could aid in the early diagnosis and prediction of thrombocytopenia severity, inform treatment strategies, and facilitate the removal of pathogenic antibodies from circulation.

1. Jacob EA. Complete blood cell count and peripheral blood film, its significant in laboratory medicine: a review study. Am J Lab Med. 2016;1(3):34-57.
2. Kazemi MH, Malakootikhah F, Momeni-Varposhti Z, et al. Human platelet antigen 1-6, 9 and 15 in the Iranian population: An anthropological genetic analysis. Sci Rep. 2020;10(1):7442.
3. Fritsma GA. Platelet structure and function. Clin Lab Sci. 2015;28(2):125-131.
4. Flaumenhaft R. Platelets: out of shape and misbehaving. Blood. 2022;140(21):2188-90.
5. Roz̆man P. Platelet antigens. The role of human platelet alloantigens (HPA) in blood transfusion and transplantation. Transpl Immunol. 2002;10(2-3):165-81.
6. Periayah MH, Halim AS, Saad AZM. Mechanism action of platelets and crucial blood coagulation pathways in hemostasis. Int J Hematol Oncol Stem Cell Res. 2017;11(4):319-327.
7. Crawley J, Zanardelli S, Chion C, et al. The central role of thrombin in hemostasis. J Thromb Haemost. 2007:5 Suppl 1:95-101.
8. Semple JW, Italiano JE, Freedman J. Platelets and the immune continuum. Nat Rev Immunol. 2011;11(4):264-74.
9. Reitsma S, oude Egbrink MG, Viviane VT, et al. Endothelial glycocalyx thickness and platelet-vessel wall interactions during atherogenesis. Thromb Haemost. 2011;106(5):939-46.
10. Rand ML, Leung R, Packham MA. Platelet function assays. Transfus Apher Sci. 2003;28(3):307-17.
11. Curtis B, McFarland J. Human platelet antigens–2013. Vox Sang. 2014;106(2):93-102.
12. Kuroishi A, Takihara Y, Hirayama F. Current understanding and future perspectives for anti‐human platelet antigen‐15 antibodies in patients with alloimmune thrombocytopenia: History, laboratory testing, and clinical impact. Transfusion. 2022; 62(5):1128-1141.
13. Allen D, Ouwehand WH, De Haas M, et al. Interlaboratory variation in the detection of HPA‐specific alloantibodies and in molecular HPA typing. Vox Sang. 2007;93(4):316-24.
14. Fullard JF. The role of the platelet glycoprotein IIb/IIIa in thrombosis and haemostasis. Curr Pharm Des. 2004;10(14):1567-76.
15. Carlsson LE, Greinacher A, Spitzer C, et al. Polymorphisms of the human platelet antigens HPA-1, HPA-2, HPA-3, and HPA-5 on the platelet receptors for fibrinogen (GPIIb/IIIa), von Willebrand factor (GPIb/IX), and collagen (GPIa/IIa) are not correlated with an increased risk for stroke. Stroke. 1997;28(7):1392-5.
16. Ferrington DA, Gregerson DS. Immunoproteasomes: structure, function, and antigen presentation. Prog Mol Biol Transl Sci. 2012:109:75-112
17. Salem AHA, Hamed AESA, Abdalla EM, et al. Gene frequencies of human platelet alloantigens 1–5 in two Arab populations. Blood Transfus. 2014;12 Suppl 1(Suppl 1):s281-5.
18. Song KS, Kim BS, Choi CR, et al. Association of Br polymorphism of platelet GP Ia gene and immune thrombocytopenic purpura. Platelets. 1997;8(5):361-5.
19. Najaoui A, Bakchoul T, Stoy J, et al. Autoantibody‐mediated complement activation on platelets is a common finding in patients with immune thrombocytopenic purpura (ITP). Eur J Haematol. 2012;88(2):167-74.
20. Koutsogianni P. Nomenclature of human platelet antigens and clinical conditions. Haematol. 2004;7:S82-8.
21. Kuijpers RW, Ouwehand WH, Bleeker PM, et al. Localization of the platelet-specific HPA-2 (Ko) alloantigens on the N-terminal globular fragment of platelet glycoprotein Ib alpha. Blood. 1992;79(1):283-8.
22. Kjeldsen-Kragh J, Bengtsson J. Fetal and Neonatal Alloimmune Thrombocytopenia-New Prospects for Fetal Risk Assessment of HPA-1a-Negative Pregnant Women. Transfus Med Rev. 2020;34(4):270-276.
23. Ertel K, Al‐Tawil M, Santoso S, et al. Relevance of the HPA‐15 (Gov) polymorphism on CD109 in alloimmune thrombocytopenic syndromes. Transfusion. 2005;45(3):366-73.
24. Edinur H, Dunn P, Lea R, et al. Human platelet antigens frequencies in Maori and Polynesian populations. Transfus Med. 2013;23(5):330-7.
25. Phuangtham R, Romphruk A, Puapairoj C, et al. Human platelet antigens in Burmese, Karen and north-eastern Thais. Transfus Med. 2017;27(1):60-5.
26. Thude H, Gatzka E, Anders O, et al. Allele frequencies of human platelet antigen 1, 2, 3, and 5 systems in patients with chronic refractory autoimmune thrombocytopenia and in normal persons. Vox Sang. 1999;77(3):149-53.
27. Nurden AT. Polymorphisms of human platelet membrane glycoproteins: structure and clinical significance. Thromb Haemost. 1995;74(1):345-51.
28. Castro V. Human platelet antigens and primary immune thrombocytopenia. Rev Bras Hematol Hemoter. 2017;39(2):95-97.
29. Salah M, Hamaz S, Bachir H, et al. Association between Immunological Thrombocytopenia and Thrombosis, Is It an Exotic Phenomenon?—A Case Report. J Biosci Med. 2022;10(9):289-93.
30. Hurd C, Cavanagh G, Schuh A, et al. Genotyping for platelet‐specific antigens: techniques for the detection of single nucleotide polymorphisms. Vox Sang. 2002;83(1):1-12.
31. Hayashi T, Hirayama F. Advances in alloimmune thrombocytopenia: perspectives on current concepts of human platelet antigens, antibody detection strategies, and genotyping. Blood Transfus. 2015;13(3):380-90.
32. Malakootikhah F, Naghavi H, Firouzabadi N, et al. Association of human platelet alloantigens encoding gene polymorphisms with the risk of Coronary artery disease in Iranian patients. BMC cardiovasc Disord. 2021;21(1):68.
33. Eyada TK, Amin DG, Samih I, et al. Human platelet antigen 1, 2 and 5 gene polymorphisms in Egyptians and their potential association with susceptibility to immune thrombocytopenic purpura in Egyptian patients. Hematology. 2018;23(2):111-6.
34. Naderi M, Habibpour M, Alizadeh S, et al. Study of the relationship between HPA-1 and HPA-5 gene polymorphisms and refractory to platelet therapy and recombinant factor VII in Glanzmann thrombasthenia patients in southeast of Iran. Int J Hematol Oncol Stem Cell Res. 2018;12(1):43-48.
35. Curtis BR, editor Genotyping for human platelet alloantigen polymorphisms: applications in the diagnosis of alloimmune platelet disorders. Seminars in thrombosis and hemostasis; 2008: © Thieme Medical Publishers.
36. Nomura S, Matsuzaki T, Yamaoka M, et al. Genetic analysis of HLA-and HPA-typing in idiopathic (autoimmune) thrombocytopenic purpura patients treated with cepharanthin. Autoimmunity. 1999;30(2):99-105.
37. Muhiddin R, Nurulita A, Harjianti T, et al. Analysis of Human Platelet Antigen (HPA) in Thrombocytopenia Patients. Indian J Public Health Res. 2019;10(10): 1570-1575.
Files
IssueVol 18 No 4 (2024) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijhoscr.v18i4.16762
Keywords
Genotype; Autoimmune thrombocytopenia; Platelet-specific antigens (HPA)

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
1.
Asgari Nejad S, Hashemzadeh P, Abdolkarimi B, Karimi Rouzbahani A, Anani Sarab G, Varzi AM. Association of HPA Antigens with Immune Thrombocytopenia: A Case-Control Study by PCR-SSP Method. Int J Hematol Oncol Stem Cell Res. 2024;18(4):378-390.