Blood Group Antigens Visualisation on Leukocytes
Abstract
Background/Aim: The leukocytes have been reported to contain blood group specific antigens, that are clinically relevant, however visualisation of A and B group antigens on leukocytes is a big issue. In cases of ABO discrepancies weak blood group antigens on nuclear cells have been demonstrated by using expensive techniques. Thus, the development of the method of the detection of weak blood group antigens on leukocytes available for any laboratory technician is hardly essential. The study aimed to reveal and analyse A and B blood group specific adsorbing antigens on leukocytes and erythrocytes and to develop a method for visualisation of weak blood group antigens on leukocytes.
Methods: Polyclonal and monoclonal anti-A and anti-B antibodies, received from international laboratories according to the program of Workshop IV, held in Paris, 2000, were used for the study. Mixed agglutination reaction was performed as the method for visualisation of weak blood group antigens on leukocytes as nuclear cells.
Results: Polyclonal sera from O blood group persons without weak blood group antigens in contrast to monoclonal antibodies demonstrated the ability to reveal weak blood group specific antigens on leukocytes by the method of mixed agglutination reaction. However, the test erythrocytes from the persons with increased levels of platelets and erythrocyte sedimentation rate did not allow to visualise weak antigen expression on the studied leukocytes in contrast to the persons with normal levels of platelets and erythrocyte sedimentation rate, that successfully formed mixed agglutinates with weak blood group antigens on leukocytes in mixed agglutination reaction. The leukocytes suspended in 0.9 % saline (as a diluent) incubated with the mixture of the serum with 0.9 % saline (1:2) led to the formation of specific agglutinates with test erythrocytes. The experiments with different temperature regimes and time of incubation demonstrated the usefulness of the studied method in specific leukocytes antigen visualisation during prolonged incubation at 4 °C. The persons with weak group A and B antigens, revealed on the leukocytes by the studied method, demonstrated decreased level of erythrocytes, platelets, titre of corresponding warm agglutinating antibodies (less than 1:8) and increased erythrocyte sedimentation rate.
Conclusion: The mixed agglutination reaction with prolonged incubation at 4 °C and the use of the selected polyclonal sera and test erythrocytes from the donors with normal values of platelets and erythrocyte sedimentation rate may be used for weak blood group antigens detection on leukocytes. The donors of the sera and test erythrocytes used in mixed agglutination reaction should be investigated on common blood analysis, agglutinating titre of corresponding warm group specific antibodies and presence of weak blood group antigens.
References
Ewald DR, Sumner SC. Blood type biochemistry and human disease. Wiley Interdiscip Rev Syst Biol Med. 2016;8(6):517-35. doi: 10.1002/wsbm.1355.
Wang X, Zhang F, Jiang Y, Xu Z, Feng X, Li L, ET AL. Highly individual- and tissue-specific expression of glycoprotein group A and B blood antigens in the human kidney and liver. BMC Immunol. 2021 Oct 1;22(1):66. doi: 10.1186/s12865-021-00456-2.
Mahmoud A, Comhaire F. Antisperm antibodies: Use of the mixed agglutination reaction (MAR) test using latex beads. Human Reproduction. 2000;15(2):231–3. doi: 10.1093/humrep/15.2.231.
Jager S, Kremer J, van Slochteren-Draaisma T. A simple method of screening for antisperm antibodies in the human male. Detection of spermatozoal surface IgG with the direct mixed antiglobulin reaction carried out on untreated fresh human semen. Int J Fertil. 1978;23:12-21. PMID: 30704.
Paschke R, Schulze BD, Heinecke A. Significance of sperm antibodies detected by the mixed antiglobulin reaction and the tray agglutination test. Andrologia. 1994;26:263–9. doi: 10.1111/j.1439-0272.1994.tb00800.x.
Rasanen M, Agrawal YP, Saarikoski S. Seminal fluid antisperm antibodies measured by direct flow cytometry do not correlate with those measured by indirect flow cytometry, the indirect immunobead test, and the indirect mixed antiglobulin reaction. Fertil Steril. 1996;65:170–5. doi: 10.1016/s0015-0282(16)58047-9.
Sinisi AA, Di Finizio B, Pasquali D, Scurini C, D'Apuzzo A, Bellastella A. Prevalence of antisperm antibodies by SpermMARtest in subjects undergoing a routine sperm analysis for infertility. Int J Androl. 1993 Oct;16(5):311-4. doi: 10.1111/j.1365-2605.1993.tb01197.x..
El Dewi DM, Metwally T. Adsorption Technique in pre-transfusion testing for patients with warm type autoimmune hemolytic anemia. Egypt J Immunol. 2017;24(2):47-51. PMID: 29528578.
Wright JM. The A and B blood group substances on odontogenic epithelium. J Dental Res. 2016;58(2):624-8. doi: 10.1177/00220345790580021501.
Dabelsteen E, Fejerskov O, Francois D. Ultrastructural localization of blood group antigen A and cell coat on human buccal epithelial cells. Acta Pathol Microbiol Scand B Microbiol Immunol. 1974 Feb;82(1):113-21. doi: 10.1111/j.1699-0463.1974.tb02301.x.
Kravchun PG, Leontyeva FS, Dielievska VY, Povelichenko OD. The differences in the binding of IgM and IgG antibodies with erythrocytes and epithelial cells. New Emirates Med J. 2024;5:1-7.
Brecher ME, ed. Technical manual. 14th ed. Bethesda, MD: American Association of Blood Banks, 2002: 673.
Coombs RR, Daniel MR, Gurner BW, Kelus A. Recognition of the species of origin of cells in culture by mixed agglutination. I. Use of antisera to red cells. Immunology. 1961;4(1):55-66. PMID: 13695283;
Swinburne LM, Frank BB, Coombs RRA. The A antigen on the buccal epithelial cells of man. Vox Sang. 1961;6(3):274–86. doi: 10.1159/000455443.
Pamphilon DH, Scott ML. Robin Coombs: his life and contribution to haematology and transfusion medicine. British J Haemat. 2007;137(5):401-8. doi: 10.1111/j.1365-2141.2007.06531.x.
Kravchun PG, Babadzan VD, Leontyeva FS, Dielievska VY. Detection of weak A antigen associated with Proteus vulgaris infection. J Assoc Med Sci. 2023;56(3):42-51.
Kravchun PG, Leontyeva FS Povelichenko OD, Dielievska VY. The differences of blood group typing using erythrocytes and leukocytes. New Emirates Med J. 2024;5(3):e02506882279058.
Hermelin D, Zhang D, Blackall D. Resolution of an unexpected ABO typing discrepancy in a 9-month-old patient with juvenile myelomonocytic leukemia. Clin Case Rep. 2020;8:2358–60. doi: 10.1002/ccr3.3101.
Dielievska V, Korzh M, Leontieva F, Ashukina N, Borzova O. A clinical case of weak A antigen on the erythrocytes in a person with coexistent anti-A antibodies. Arch Razi Inst. 2020;75(2):257–65. doi: 10.22092/ARI.2020.341761.1439.
Chun S, Ryu MR, Cha SY, Seo JY, Cho D. ABO mistyping of cis-AB blood group by the automated microplate technique. Transfus Med Hemotherapy. 2018;45:5–10. PMID: 29593455.
Ryu SE, Lee HJ, Kim HR, Shin KH, Kim HH. Evaluation of the automated analyzer AutoVue innova for blood group typing: clinical implementation at a tertiary hospital over 2 years. Korean J Blood Transfus. 2020;31:131–40. doi:10.17945/kjbt.2020.31.2.131.
Cooling LW, Kelly K, Barton J, Hwang D, Koerner TAW, Olson JD. Determinants of ABH expression on human blood platelets. Blood. 2005;105(8):3356–64. doi: 10.1182/blood-2004-08-3080.
Kravchun PG, Korzh MO, Zinchenko OA, Lyzohub MV, Dielievska VY. Resolving discrepancies in forward and reverse blood grouping. Scr Med. 2023;54(4):425-37. doi: 10.5937/scriptamed54-45572.
Coombs RR, Franks D, Gurner BW, Polley MJ, Richards CB. Studies on mixed agglutination with special reference to the A antigen and the physical properties of the operative antibodies. Immunology. 1965;8(2):182-92. PMID: 14295642.
Moller G, Wigzell H. Antibody synthesis at the cellular level. Antibody-induced suppression of 19S and 7S antibody response. J Exp Med. 1965;121(6):969-89. doi: 10.1084/jem.121.6.969.
Henry C, Jerne NK. Competition of 19S and 7S antigen receptors in the regulation of the primary immune response. J Exp Med. 1968;128(1):133-52. doi: 10.1084/jem.128.1.133.
Pearlman DS. The influence of antibodies on immunologic responses. I. The effect on the response to particulate antigen in the rabbit. J Exp Med. 1967;126(1):127-48. doi: 10.1084/jem.126.1.127.
Hjelm F, Carlsson F, Getahun A, Heyman B. Antibody-mediated regulation of the immune response. J Immunol. 2006;64(3):177-84. doi: 10.1111/j.1365-3083.2006.01818.x.
Macagno S, Smedile A, Caredda F, Ottobrelli A, Rizzetto M. Monomeric (7S) immunoglobulin M antibodies to hepatitis delta virus in hepatitis type D. Gastroenterology. 1990;98(6):1582-6. doi: 10.5555/uri:pii:001650859091094M.
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).