Abstract
This study aimed to characterize anti‐A and anti‐B hemolysins, IgM, and IgG titers in Thai blood donors. Altogether, 300 serum samples from group O donors at the National Blood Centre, Thai Red Cross Society, were screened for anti‐A and anti‐B hemolysins and treated with 0.01 M dithiothreitol to characterize IgM and IgG titers by standard tube technique. Antibody titers were compared with hemolysis grade. Male and female ratio = 1:1.3 and ages ranged from 17 to 60 years. The overall prevalence of anti‐A and anti‐B hemolysins was 69%. Anti‐A and anti‐B hemolysins comprised 18.3% and 16.7%, respectively and 34% had both antibodies. High titers of anti‐A hemolysins were associated with females (P< 0.05), and only anti‐B IgM titers were associated with age (P< 0.05). Interestingly, the association of anti‐A IgM titers, anti‐A IgG titers, and hemolysin grade was demonstrated (P< 0.05). A significant association between hemolysin grade and anti‐B IgM titers was found (P< 0.05). The prevalence of anti‐A and anti‐B hemolysins and high titers of IgM and IgG in Thais are high. Hemolysin grade showed significant associations with IgM titers; therefore, when providing ABO‐incompatible platelet transfusion, especially for female plateletpheresis donors, IgM high titers of anti‐A and anti‐B screening is suggested. J. Clin. Lab. Anal. 26:223‐226, 2012. © 2012 Wiley Periodicals, Inc.
Keywords: ABO antibodies, hemolysins, IgM and IgG titers, group O, Thais
INTRODUCTION
ABO compatibility and platelet transfusion is clinically important because platelet increments are decreased after transfusion of ABO incompatible platelets 1. Additionally, acute hemolytic transfusion reactions, venoocclusive disease, and increased morbidity in allogeneic transplants caused by minor‐ABO incompatible platelet transfusions have been reported 2, 3, 4, 5, 6, 7. Recent studies showed that approximately 10% to 40% of group O donors have high titer of IgG anti‐A and anti‐B 1, 8, 9, and not only IgM and IgG titers but also anti‐A and anti‐B hemolysins, which are IgG can cause hemolysis of red blood cells when group O blood/plasma is transfused to nongroup O patients and cause hemolytic disease of the fetus and newborn 10. A case of severe fetal anemia due to ABO incompatibility caused by high anti‐B IgG titers of African mother was reported 11. The high frequency of hemolysins in group O blood donors was commonly found in Nigerian and Indian populations with the prevalence ranging from 23.2% to 62.8% 12, 13, 14, 15. A previous study of ABO antibody levels among Japanese, Lao, and Thai blood donors in 2006 showed that IgM anti‐A and anti‐B titers were low in Thais, similar to the Japanese antibody titers reported in 2001, but IgG anti‐A and anti‐B titers were high, similar to those reported in Japanese in 1986 16. According to the requirements of the Standards for Blood Banks and Transfusion Services, it is recommended that the blood bank have a policy concerning transfusion of components containing significant amounts of ABO incompatible antibodies 17. Screening for donor anti‐A and anti‐B hemolysins, as well as high titers of IgM and IgG is suggested when using platelets containing ABO incompatible plasma 18. Since information on immunoglobulin types and concentrations of ABO hemolysins in the Thai population is still unclear, the purpose of this study was to characterize anti‐A and anti‐B hemolysins, IgM, and IgG titers in group O Thai blood donors.
MATERIALS AND METHODS
Subjects
Altogether, 300 blood samples obtained from group O Thai blood donors of the National Blood Centre, Thai Red Cross Society, Bangkok, Thailand were included in this study. There were 128 males and 172 females (M:F = 1:1.3) and their ages ranged from 17 to 60 years. Informed consent was obtained from each subject. This study was approved by the Committee on Human Rights Related to Research Involving Human Subjects, Thammasat University, Pathumtani, Thailand.
Methods
ABO blood group was determined by cell and serum grouping according to methods previously described 19. Anti‐A and anti‐B antiserum for cell grouping were obtained from the National Blood Centre, Thai Red Cross Society and pooled cells for serum grouping were prepared in‐house. Test for hemolysins and scoring was performed by standard procedure 20. Hemolysin was graded as follows: 3+ (complete hemolysis), and 2+ (partial > 50% but not complete), 1+ (trace hemolysis), and negative (no visible sign of hemolysis). Serum samples having a score of 3+ and 2+ were considered strongly hemolytic. IgM anti‐A and anti‐B titers were determined using the saline method with incubation at room temperature for 5 min. The IgG antibody titers were determined by an indirect antiglobulin test (IAT) and an incubation of 30 min at 37°C. Before the IAT, the sera were treated with 0.01 M dithiothreitol for 15 min at 37°C using the method as described by Brecher 21, to destroy IgM antibodies in their sera. The results of hemolysins, titers for IgM, and IgG ABO antibody testing can be obtained within 20 min, 10 min, and 60 min, respectively.
Statistical Analysis
Chi‐square test was used to determine the association between grade of hemolysins and IgM and IgG antibody titers. A level of P < 0.05 was accepted as significant.
Results
The distribution of male and female donors among different age groups (≤30, 31–40, 41–50, and >50 years) showed significant association (P < 0.05). The highest numbers of female donors (41.9%) was found in the younger than 30 age group, while that of male blood donors was 30.5% in the 41 to 50 year group, as shown in Table 1.
Table 1.
Distribution of Age and Sex in 300 Blood Donors
| Age (year) | Sex | Total (%) | |
|---|---|---|---|
| Male (%) | Female (%) | ||
| ≤ 30 | 36 (28.1) | 72 (41.9) | 108 (36.0) |
| 31–40 | 36 (28.1) | 49 (28.5) | 8.5 (28.3) |
| 41–50 | 39 (30.5) | 38 (22.1) | 77 (25.7) |
| >51 | 17 (13.3) | 13 (7.6) | 30 (10.0) |
| Total | 128 (100.0) | 172 (100.0) | 300 (100.0) |
Hemolysins
The overall prevalence of anti‐A and anti‐B hemolysins in serum samples from group O Thai blood donors was 69%. Prevalence of anti‐A and anti‐B hemolysins was 18.3% and 16.7%, respectively, while that of donors having both anti‐A and anti‐B hemolysins in their sera was 34%. Anti‐A and anti‐B hemolysins were not associated with age, while high titers of anti‐A hemolysins were associated with females (P < 0.05).
IgM Titers
In this study, IgM anti‐A and anti‐B titers in group O blood donors ranged from 8 to 1,024 (IgM anti‐A titer ≥64 was 75.7% and IgM anti‐B titer ≥64 was 80.0%), and no association was noted between the sexes. Only a decrease of anti‐B IgM titers was associated with an increase of donor age in both males and females (P < 0.05).
IgG Titers
Regarding IgG anti‐A and anti‐B titers, the distribution was between 16 and 4,096 (IgG anti‐A titer ≥64 was 93.0% and IgG anti‐B titer ≥64 was 95.3%) and no association was found between sex and age.
Association of Hemolysins, IgM Titers, and IgG Titers
Interestingly, the association among anti‐A IgM titers, anti‐A IgG titers, and anti‐A hemolysin grade were demonstrated (P < 0.05), whereas significant association was found (P < 0.05) between anti‐B hemolysin grade and anti‐B IgM titer, as shown in Table 2.
Table 2.
Comparison of Hemolysin Grades and Immunoglobulin Titers
| Anti‐A titer | Anti‐B titer | |||||||
|---|---|---|---|---|---|---|---|---|
| IgM[Link] | IgG | IgM[Link] | IgG | |||||
| Hemolysin grade/titer | <64 | ≥64 | <64 | ≥64 | <64 | ≥64 | <64 | ≥64 |
| 0 and 1 | 44.8% | 55.2% | 9.0% | 91.0% | 33.8% | 66.2% | 6.8% | 93.2% |
| 2 & 3 | 5.2% | 94.8% | 5.2% | 94.8% | 6.6% | 93.4% | 2.6% | 97.4% |
*P value < 0.05.
DISCUSSION
It is suggested that environmental factors, enteric bacteria, and other parasites may have affected the production of anti‐A and anti‐B titers 22, 23, 24. Moreover, a high frequency of strongly hemolytic anti‐A and anti‐B hemolysins has been reported in Asian and Black populations compared with Caucasians 12, 23, 25, which may be due to mosquito bites and intestinal parasitic infections 15. In addition, high titers of ABO antibodies in group O individuals can also be the outcome of vaccination or other antigen exposures 26.
The data on the Ig types and concentration of ABO antibodies in the Thai population are obscure. The high frequency of anti‐A and anti‐B hemolysins in group O blood donors in this study is higher than other previous studies in Nigerian and Indian populations because of the increasing numbers of female donors. High titers of anti‐A hemolysins were also associated with female blood donors, which may be caused by pregnancies.
A previous study in Japanese group O blood donors showed that IgM anti‐A, and anti‐B titers were low, and no differences were noted between sexes 16, while in this study IgM anti‐A and anti‐B titers in group O Thai blood donors were high. Only IgM anti‐B titers were significantly decreased according to an increase of donor age.
Additionally, IgG titers of anti‐A and anti‐B in Japanese donors showed differences between sexes and increase in donor age, especially in female blood donors 16. In contrast, no association was found between sex and age with IgM and IgG anti‐A and anti‐B titers in group O Thai blood donors.
Correlation between IgM and IgG levels has been demonstrated in Zimbabweans 25. We also found significant association among anti‐A IgM titers, anti‐A IgG titers, and anti‐A hemolysin grade, while only significant association between anti‐B hemolysin grade and anti‐B IgM titer was demonstrated.
In cases of emergency, it is common practice to transfuse ABO‐incompatible platelets to patients because of limited supply and short shelf life of platelets. Therefore, it is important to avoid transfusion of immune anti‐A and anti‐B antibodies to nongroup O patients. This study reports a high frequency of immune ABO antibodies in group O Thai blood donors. In addition to screening for hemolysins and high IgG titers, which is labor‐intensive and time‐consuming, screening for high IgM titers is beneficial because of its simplicity. Therefore, it can be used as an alternative to screen female group O apheresis donors.
ACKNOWLEDGMENTS
This work is supported by the National Research University Project of Thailand, Office of Higher Education Commission, and the Faculty of Allied Health Sciences, Thammasat University. We also thank Mr. Jesada Kittikul for his advice concerning statistics.
REFERENCES
- 1. Fung MK, Downes KA, Shulman IA. Transfusion of platelets containing ABO‐incompatible plasma: a survey of 3156 North American laboratories. Arch Pathol Lab Med 2007;131:909–916. [DOI] [PubMed] [Google Scholar]
- 2. Cooling L. ABO and platelet transfusion therapy. Immunohematol 2007;23:20–33. [PubMed] [Google Scholar]
- 3. McManigal S, Sims KL. Intravascular hemolysis secondary to ABO incompatible platelet products: An under recognized transfusion reaction. Am J Clinical Pathol 1999;111:202–206. [DOI] [PubMed] [Google Scholar]
- 4. Larsson LG, Welsh VJ, Ladd DJ. Acute intravascular hemolysis secondary to out‐of group platelet transfusion. Transfusion 2000;40:902–906. [DOI] [PubMed] [Google Scholar]
- 5. Herman JH. Apheresis platelet transfusions: Does ABO matter? Transfusion 2004;44:802–804. [DOI] [PubMed] [Google Scholar]
- 6. Benjamin RJ, Antin JH. ABO‐incompatible bone marrow transplantation: The transfusion of incompatible plasma may exacerbate regimen‐related toxicity. Transfusion 1999;39:1273–1274. [DOI] [PubMed] [Google Scholar]
- 7. Lapierre V, Mahe C, Auperin A, et al. Platelet transfusion containing AB‐incompatible plasma and hepatic veno‐occlusive disease after hematopoietic transplantation in young children. Transplantation 2005;80:314–319. [DOI] [PubMed] [Google Scholar]
- 8. Josephson CD, Mullis NC, Van Demark C, Hillyer CD. Significant numbers of apheresis‐derived group O platelet units have “high‐titer” anti‐A/A,B: Implications for transfusion policy. Transfusion 2004;44:805–808. [DOI] [PubMed] [Google Scholar]
- 9. Harris SB, Josephson CD, Kost CB, Hillyer CD. Nonfatal intravascular hemolysis in a pediatric patient after transfusion of a platelet unit with high‐titer anti‐A . Transfusion 2007;47:1412–1417. [DOI] [PubMed] [Google Scholar]
- 10. Boorman KE, Dodd BE, Lincoln PJ, eds. Blood Group Serology, sixth edition Edinburgh, UK: Churchill Livingstone; 1988. [Google Scholar]
- 11. Ziprin JH, Payne E, Hamidi L, Roberts I, Regan F. ABO incompatibility due to immunoglobulin G anti‐B antibodies presenting with severe fetal anaemia. Transfus Med 2005;15:57–60. [DOI] [PubMed] [Google Scholar]
- 12. Kullarni AG, Ibazebe R, Fleming AF. High frequency of anti‐A and anti‐B haemolysins in certain ethnic group of Nigeria. Vox Sang 1985;48:39–41. [DOI] [PubMed] [Google Scholar]
- 13. Olawumi HO, Olatunji PO. Prevalence and titre of alpha and beta haemolysins in blood group O donors in Ilorin. Afr J Med Sci 2001;30:319–321. [PubMed] [Google Scholar]
- 14. Kagu MB, Ahmed SG, Mohammed AA, Moshood WK, Malah MB, Kehinde MJ. Anti‐A and anti‐B haemolysins among group “O” voluntary blood donors in Northern Nigeria. J Transfus 2011; Article ID 302406, doi: 10.4061/2011/302406. [DOI] [Google Scholar]
- 15. Mathai J, Sinthu PV, Sulochana PV, Sathyabhama S. Haemolysin test for characterization of immune ABO antibodies. Indian J Med Res 2003;118:125–128. [PubMed] [Google Scholar]
- 16. Mazda T, Yabe R, Nathalang O, Thammavong T, Tadokoro K. Differences in ABO antibody levels among blood donors: A comparison between past and present Japanese, Laotians, and Thai populations. Immunohematol 2007;23:38–41. [PubMed] [Google Scholar]
- 17. American Association of Blood Banks , ed. Standards for Blood Banks and Transfusion Services, 26th edition Bethesda, MD: American Association of Blood Banks; 2009. [Google Scholar]
- 18. Pietersz RNI, Engelfriet CP, Reesink HW. International forum: Transfusion of apheresis platelets and ABO groups. Vox Sang 2005;88:207–221. [DOI] [PubMed] [Google Scholar]
- 19. Roback JD, Combs MR, Grossman BJ, Hillyer CD, eds. Technical Manual, 16th edition Bethesda, MD, American Association of Blood Banks; 2008. [Google Scholar]
- 20. Boorman KE, Dodd BE, Lincoln PJ, eds. Blood Group Serology: Theory, Techniques, Practical Applications, fifth edition Edinburgh, UK: Churchill Livingstone; 1977. [Google Scholar]
- 21. Brecher ME, ed. Method 2.11. Use of Sulfhydryl Reagents to Disperse Autoagglutination Technical Manual, 15th edition Bethesda, MD: American Association of Blood Banks; 2005. [Google Scholar]
- 22. Toy PTCY, Reid ME, Papenfus L, Yeap HH, Black D. Prevalence of ABO maternal‐infant incompatibility in Asians, Blacks, Hispanics and Caucasians. Vox Sang 1988;54:181–183. [DOI] [PubMed] [Google Scholar]
- 23. Redman M, Malde R, Contreras M. Comparison of IgM and IgG anti‐A and anti‐B levels in Asian, Caucasian and Negro donors in the North West Thames region. Vox Sang 1990;59:89–91. [DOI] [PubMed] [Google Scholar]
- 24. Issitt PD, Anstee DJ, eds. Applied Blood Group Serology, fourth edition Durham, NC: Montgomery Scientific Publications; 1998. [Google Scholar]
- 25. Adewuyi JO, Gwanzura C, Mvere D. Characteristics of anti‐A and anti‐B in black Zimbabweans. Vox Sang 1994;67:307–309. [DOI] [PubMed] [Google Scholar]
- 26. Siber GR, Ambrosino DM, Gorgone BC. Blood‐group‐A like substance in a preparation of pneumococcal vaccine. Ann Intern Med 1982;96:580–586. [DOI] [PubMed] [Google Scholar]