GENOTYPING OF AB0 AND Rh SYSTEMS BLOOD GROUPS IN PATIENTS AFTER MULTIPLE HEMOTRANSFUSIONS

Background. when transplantation of hematopoietic stem cells (HSC) is performing, it is necessary to take into account the incompatibility of the donor and recipient in terms of erythrocyte antigens in order to assess the possibility of immunological complications during HSC transfusion and/or graft engraftment (acute hemolysis, delayed hemolysis, etc.). The results of serological research methods do not always allow identifying the true group affiliation due to posttransfusion chimerism in patients and/or the presence of antigen allelic polymorphism.Aim. To establish the frequency of ABO-incompatible allo-HSC transplantations in the National Research Center for Hematology, to determine by molecular methods the group affiliation of patients with a weakened expression of antigens and/or after multiple blood transfusions before HSC transplantation, to clarify the blood type of HSC donors with a weakened expression of antigens.Materials and methods. The blood of 270 HSC donor-recipient couples was examined. The blood group of the ABO, Rhesus, MNS, Kell systems was determined in a plane agglutination test using the corresponding IgM class Tsoliclones and in gel cards. Genotyping was performed by polymerase chain reaction with primers to identify the genes of the ABO, Rhesus, Kell, and MNS systems.Results. In 2018-2020 270 HSC transplantations were performed at the National Research Center for Hematology. In 141 (52.22 %) couples, incompatibility of the donor and recipient according to the ABO system was revealed: major - 23.33 %, minor - 20 %; bidirectional - 8.89 %. problems in assessing of serological results were observed in 97 (36.3 %) patients: in 78 patients with post-transfusion chimerism and 19 patients with weakened antigen expression; in 15 (5.56 %) HSC donors: in 4 due to the lack of information about the blood group of cryopreserved cells, in 10 due to weakened antigen expression, in 1 to search for informative markers for monitoring HSC engraftment. The results of the study demonstrated that the percentage of agglutinated erythrocytes in post-transfusion chimerism cannot be a reliable criterion for establishing the true phenotype of a patient. In donors and patients with weakened expression of antigens, the presence of ABO*O1, -A1, -A2, -B1, RHD weak type 1, RHD weak type 2, RHD weak type 3, RHCE*Cw genes was confirmed. for the first time in Russia gene RHCE*01.38 was found.Conclusion. The prevalence of ABO-incompatible HSC transplants was noted. problems with serological determination of the blood group in a third of patients before HSC transplantation arose due to the presence of post-transfusion chimerism and weakened expression of antigens. Determining of the genotypes of HSC donors is necessary when the expression of antigens is weakened and cryopreserved cells are received. The percentage of agglutinated erythrocytes in post-transfusion chimerism cannot be a reliable criterion for establishing the true phenotype of a patient. Detection of mixed chimerism in the determination of group factors by serological methods is an indication for genotyping, especially in the context of the predominance of incompatible HSC transplantations.

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Molecular methods in identifying the true grouping of erythrocytes in patients with blood system diseases before transplantation of hematopoietic stem cells and their donors

Головкина

Стремоухова

Пушкина

et al. 2022

Onkogematologiâ

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Blood group genotyping in multi-transfused patients

et al. 2021

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Aim. To assess the possibility of using blood group genotyping in recipients who received transfusions for 3 months. Methods. The study included blood samples from 95 patients who received 3 or more erythrocyte transfusions within 3 months. The patients had the following diagnoses: multiple myeloma (n=7), beta thalassemia (n=4), non-Hodgkin's lymphomas (n=11), chronic myeloid leukemia (n=16), primary myelofibrosis (n=9), myelodysplastic syndrome (n=22), acute leukemia (n=21), aplastic anemia (n=5). Red blood cells phenotyping was performed in Diaclon Rh Subgroups+K Gel Cards. The Rh and Kell genotyping was performed by using RBC SSP-PCR kits FluoGene vERYfy (Inno-train Diagnostics, Germany). The standard RHD/RHCE alleles, as well as polymorphisms associated with KEL1/KEL2 [T698C (Met198Thr)] of the KEL gene were genotyped. Results. The concordance rate between serological and molecular genetic typing of RhCE and Kell blood groups for donors was 100%, while the patients results were discordant in 45.3% of cases. Discrepancies in antigens of the Rh system were registered in 41 patients: one antigen of the Rh system in 30 patients, two in 9 patients. Ten patients who had been previously phenotyped as RhCc were genotyped as RHCE*CC. 2 patients who had been previously phenotyped as Rhee were genotyped as RHCE*EE. In 2 patients, antigens D and C were not detected in the phenotype but were identified in the genotype. Discrepancies in antigen K were recorded in 2 patients, and the antigen was absent in the phenotype but was present in the genotype. The genotyping results were confirmed by serological typing at subsequent hospitalizations. Сonclusion. Blood group genotyping is a useful adjunct to traditional methods when serological typing is limited.

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Features of immunohematological and hematological parameters of a donor with a rare phenotype -D-

Ламзин

Sokolova

Хайруллин

et al. 2020

Gematologiâ i transfuziologiâ

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Background. This study is devoted to a rare variation of the -D- phenotype . The -D- phenotype was ﬁrst discovered by R. Race, R. Sanger and J.G. Selwyn in 1951. In Russia, the phenotype -D- was ﬁrst discovered by V. Morokov in 1985. Typically, the -D- phenotype is detected when physicians examine post-transfusion complications or hemolytic disease of the newborn, since such patients demonstrate high antibody titres to absent antigens. In the present study, the -D- phenotype was detected in a primary blood donor at the clinical laboratory of the Ulyanovsk Regional Blood Transfusion Station (Ulyanovsk, Russia). Aim. To study speciﬁc features of immunohematological and hematological blood parameters in a donor with a rare variation of the -D- phenotype.Materials and methods. The detection of the -D- phenotype by immunohematological methods was carried out using automatic analysers. Molecular DNA typing was used to conﬁrm the -D- phenotype. The shape of erythrocytes of the donor with the -D- phenotype was evaluated using an atomic force microscope. The characteristics of the erythroid lineage were studied using an automatic hematological analyser.Results. The -D- phenotype was detected in a primary blood donor. Due to the extreme rarity of the -D- phenotype and the lack of programmed algorithms, the validation of the results by automatic analysers was incorrect. Of critical importance was the visual assessment of gel ID cards by the medical staff. Genotyping conﬁrmed the lack of C, c, E, e, Cw speciﬁcities in the RHCE gene. The hematological parameters of the donor were within the age norm. An assessment of the image of a cytological blood preparation did not reveal changes in the shape of erythrocytes and their size.Conclusions. The primary determination of the -D- phenotype using automatic immunohematological analysers can be complicated by the impossibility of validating the results, the incorrect operation of the installed software and the need for expert evaluation of blood samples by the staff. The presented case of the -D- phenotype was not associated with changes in the shape of erythrocytes and blood hematological parameters.

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GENOTYPING OF AB0 AND Rh SYSTEMS BLOOD GROUPS IN PATIENTS AFTER MULTIPLE HEMOTRANSFUSIONS

Cited by 3 publications

References 27 publications

Molecular methods in identifying the true grouping of erythrocytes in patients with blood system diseases before transplantation of hematopoietic stem cells and their donors

Molecular methods in identifying the true grouping of erythrocytes in patients with blood system diseases before transplantation of hematopoietic stem cells and their donors

Blood group genotyping in multi-transfused patients

Features of immunohematological and hematological parameters of a donor with a rare phenotype -D-

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