ERYTHROCYTE'S GENOMIC IN RUSSIAN MULTITRANSFUSED HEMATOLOGICAL PATIENTS
(Abstract release date: 05/19/16)
EHA Library. Golovkina L. 06/09/16; 135118; PB2218
Dr. Larisa Golovkina
Contributions
Contributions
Abstract
Abstract: PB2218
Type: Publication Only
Background
The patients suffering with ineffective hematopoiesis and peripheral cytopenias need frequent RBC transfusions. There are minimum two populations of red blood cells (RBC) in transfused within the four previous month patients – own and donor’s. This situation is called posttransfusion chimerism and serological blood typing becomes unreliable. It’s known about the immunogenicity of such blood group antigens as A, B, D, c, C, E. e in antigen-negative patients. DNA analysis is available for clinical use in various situations, especially in identification of blood group antigens in multitransfused hematological patients.
Aims
true blood group identification in multitransfused hematological patients
Methods
The blood samples drawn into citrate, EDTA anticoagulant were serotyped for RhD, Rh C/c and RhE/e antigens by anti-D, anti-C, anti-Cw, anti-c, anti-E and anti-e monoclonal antibodies (Moabs) (Hematolog, Russia). Hemagglutination typing in gel columns ID-Cards “DiaClon ABO/D+Reverse Grouping” and “DiaClon Rh-subgroups+K” (Bio-Rad, Switzerland) were used for confirmation of mixed chimerism in transfused patients: agglutinated cells forming a red line on the surface of the gel and no agglutinated cells formed compact button on the bottom of the microtube. DNA extraction was followed by commercial PCR-SSP kits with primers for typing ABO, RHD, and RHCE (BAG, Germany).
Results
We serotyped blood samples from 2055 Russians hematological patients (HP) in 2015. The problems in blood group (BG) identification for ABO system were caused by posttransfused chimerism (PTCh) in 32 primary patients (1,6%) and for Rhesus system – in 64 primary patients (3,1%). 24 patients provided informed consent for genotyping. All patients suffered with idiopathic or symptomatic cytopenia or separate anemia (aplastic anemia -5, different forms of acute leukemia -3, myelodisplastic syndrome -4, multiple myeloma – 2, β-thalassemia -1, PNH – 1, unidentified anemia – 6, partial red cell aplasia – 1, with renal transplant -1) and were transfused before hospitalization to National Research Centre for Hematology. Two patients had a problem with identification in ABO BG: 50% chimerism was detected with anti-A and anti-B Moabs in one and 50% no agglutinated RBC with anti-A – in another one. Genotyping revealed B1B1 and A1B1 BG in these patients. We couldn’t identified Rhesus phenotype in 23 HP: RhC – in 17 (percent of agglutinated RBC ranged from 40 to 90), Rhc – in 13 (agglutinated RBC 50%>90%), RhD – in 4 (agglutinated RBC 20%>90%), RhE – in 18 (agglutinated RBC 30%>90%), Rhe – in 7 (agglutinated RBC 50%>95%), RhCw – in 3 (agglutinated RBC 5%>50%). So PTCh for 1 Rh antigen was detected in 5 HP, for 2 Rh antigens – in 7, for 3 – in 6, for 4 – in 5 and for 5 Rh antigens – in one. The presence of RhCw was confirmed in two HP with 50% RhCw chimerism by genotyping. Molecular method predicted the presence of RhC in 5 HP with chimerism from 20% to 95%, Rhc – in 9 HP (chimerism ranged from 40% to 90%), RhD – in 3 (chimerism ranged from 20% to 90%), RhE – in 9 (chimerism ranged from 30% to 90%) and Rhe – in 3 HP with chimerism 95%. Monitorig of BG in all HP after four month
Conclusion
The determination of ABO and Rhesus phenotypes is very important for previously transfused patients and for patients with known irregular antibodies. The erythrocyte’s genomics has enabled the application of molecular methods to transfusion medicine and allows preventing the alloimmunization to the most clinically significant blood group antigens.
Session topic: E-poster
Keyword(s): Anemia, Chimerism, Transfusion
Type: Publication Only
Background
The patients suffering with ineffective hematopoiesis and peripheral cytopenias need frequent RBC transfusions. There are minimum two populations of red blood cells (RBC) in transfused within the four previous month patients – own and donor’s. This situation is called posttransfusion chimerism and serological blood typing becomes unreliable. It’s known about the immunogenicity of such blood group antigens as A, B, D, c, C, E. e in antigen-negative patients. DNA analysis is available for clinical use in various situations, especially in identification of blood group antigens in multitransfused hematological patients.
Aims
true blood group identification in multitransfused hematological patients
Methods
The blood samples drawn into citrate, EDTA anticoagulant were serotyped for RhD, Rh C/c and RhE/e antigens by anti-D, anti-C, anti-Cw, anti-c, anti-E and anti-e monoclonal antibodies (Moabs) (Hematolog, Russia). Hemagglutination typing in gel columns ID-Cards “DiaClon ABO/D+Reverse Grouping” and “DiaClon Rh-subgroups+K” (Bio-Rad, Switzerland) were used for confirmation of mixed chimerism in transfused patients: agglutinated cells forming a red line on the surface of the gel and no agglutinated cells formed compact button on the bottom of the microtube. DNA extraction was followed by commercial PCR-SSP kits with primers for typing ABO, RHD, and RHCE (BAG, Germany).
Results
We serotyped blood samples from 2055 Russians hematological patients (HP) in 2015. The problems in blood group (BG) identification for ABO system were caused by posttransfused chimerism (PTCh) in 32 primary patients (1,6%) and for Rhesus system – in 64 primary patients (3,1%). 24 patients provided informed consent for genotyping. All patients suffered with idiopathic or symptomatic cytopenia or separate anemia (aplastic anemia -5, different forms of acute leukemia -3, myelodisplastic syndrome -4, multiple myeloma – 2, β-thalassemia -1, PNH – 1, unidentified anemia – 6, partial red cell aplasia – 1, with renal transplant -1) and were transfused before hospitalization to National Research Centre for Hematology. Two patients had a problem with identification in ABO BG: 50% chimerism was detected with anti-A and anti-B Moabs in one and 50% no agglutinated RBC with anti-A – in another one. Genotyping revealed B1B1 and A1B1 BG in these patients. We couldn’t identified Rhesus phenotype in 23 HP: RhC – in 17 (percent of agglutinated RBC ranged from 40 to 90), Rhc – in 13 (agglutinated RBC 50%>90%), RhD – in 4 (agglutinated RBC 20%>90%), RhE – in 18 (agglutinated RBC 30%>90%), Rhe – in 7 (agglutinated RBC 50%>95%), RhCw – in 3 (agglutinated RBC 5%>50%). So PTCh for 1 Rh antigen was detected in 5 HP, for 2 Rh antigens – in 7, for 3 – in 6, for 4 – in 5 and for 5 Rh antigens – in one. The presence of RhCw was confirmed in two HP with 50% RhCw chimerism by genotyping. Molecular method predicted the presence of RhC in 5 HP with chimerism from 20% to 95%, Rhc – in 9 HP (chimerism ranged from 40% to 90%), RhD – in 3 (chimerism ranged from 20% to 90%), RhE – in 9 (chimerism ranged from 30% to 90%) and Rhe – in 3 HP with chimerism 95%. Monitorig of BG in all HP after four month
Conclusion
The determination of ABO and Rhesus phenotypes is very important for previously transfused patients and for patients with known irregular antibodies. The erythrocyte’s genomics has enabled the application of molecular methods to transfusion medicine and allows preventing the alloimmunization to the most clinically significant blood group antigens.
Session topic: E-poster
Keyword(s): Anemia, Chimerism, Transfusion
Abstract: PB2218
Type: Publication Only
Background
The patients suffering with ineffective hematopoiesis and peripheral cytopenias need frequent RBC transfusions. There are minimum two populations of red blood cells (RBC) in transfused within the four previous month patients – own and donor’s. This situation is called posttransfusion chimerism and serological blood typing becomes unreliable. It’s known about the immunogenicity of such blood group antigens as A, B, D, c, C, E. e in antigen-negative patients. DNA analysis is available for clinical use in various situations, especially in identification of blood group antigens in multitransfused hematological patients.
Aims
true blood group identification in multitransfused hematological patients
Methods
The blood samples drawn into citrate, EDTA anticoagulant were serotyped for RhD, Rh C/c and RhE/e antigens by anti-D, anti-C, anti-Cw, anti-c, anti-E and anti-e monoclonal antibodies (Moabs) (Hematolog, Russia). Hemagglutination typing in gel columns ID-Cards “DiaClon ABO/D+Reverse Grouping” and “DiaClon Rh-subgroups+K” (Bio-Rad, Switzerland) were used for confirmation of mixed chimerism in transfused patients: agglutinated cells forming a red line on the surface of the gel and no agglutinated cells formed compact button on the bottom of the microtube. DNA extraction was followed by commercial PCR-SSP kits with primers for typing ABO, RHD, and RHCE (BAG, Germany).
Results
We serotyped blood samples from 2055 Russians hematological patients (HP) in 2015. The problems in blood group (BG) identification for ABO system were caused by posttransfused chimerism (PTCh) in 32 primary patients (1,6%) and for Rhesus system – in 64 primary patients (3,1%). 24 patients provided informed consent for genotyping. All patients suffered with idiopathic or symptomatic cytopenia or separate anemia (aplastic anemia -5, different forms of acute leukemia -3, myelodisplastic syndrome -4, multiple myeloma – 2, β-thalassemia -1, PNH – 1, unidentified anemia – 6, partial red cell aplasia – 1, with renal transplant -1) and were transfused before hospitalization to National Research Centre for Hematology. Two patients had a problem with identification in ABO BG: 50% chimerism was detected with anti-A and anti-B Moabs in one and 50% no agglutinated RBC with anti-A – in another one. Genotyping revealed B1B1 and A1B1 BG in these patients. We couldn’t identified Rhesus phenotype in 23 HP: RhC – in 17 (percent of agglutinated RBC ranged from 40 to 90), Rhc – in 13 (agglutinated RBC 50%>90%), RhD – in 4 (agglutinated RBC 20%>90%), RhE – in 18 (agglutinated RBC 30%>90%), Rhe – in 7 (agglutinated RBC 50%>95%), RhCw – in 3 (agglutinated RBC 5%>50%). So PTCh for 1 Rh antigen was detected in 5 HP, for 2 Rh antigens – in 7, for 3 – in 6, for 4 – in 5 and for 5 Rh antigens – in one. The presence of RhCw was confirmed in two HP with 50% RhCw chimerism by genotyping. Molecular method predicted the presence of RhC in 5 HP with chimerism from 20% to 95%, Rhc – in 9 HP (chimerism ranged from 40% to 90%), RhD – in 3 (chimerism ranged from 20% to 90%), RhE – in 9 (chimerism ranged from 30% to 90%) and Rhe – in 3 HP with chimerism 95%. Monitorig of BG in all HP after four month
Conclusion
The determination of ABO and Rhesus phenotypes is very important for previously transfused patients and for patients with known irregular antibodies. The erythrocyte’s genomics has enabled the application of molecular methods to transfusion medicine and allows preventing the alloimmunization to the most clinically significant blood group antigens.
Session topic: E-poster
Keyword(s): Anemia, Chimerism, Transfusion
Type: Publication Only
Background
The patients suffering with ineffective hematopoiesis and peripheral cytopenias need frequent RBC transfusions. There are minimum two populations of red blood cells (RBC) in transfused within the four previous month patients – own and donor’s. This situation is called posttransfusion chimerism and serological blood typing becomes unreliable. It’s known about the immunogenicity of such blood group antigens as A, B, D, c, C, E. e in antigen-negative patients. DNA analysis is available for clinical use in various situations, especially in identification of blood group antigens in multitransfused hematological patients.
Aims
true blood group identification in multitransfused hematological patients
Methods
The blood samples drawn into citrate, EDTA anticoagulant were serotyped for RhD, Rh C/c and RhE/e antigens by anti-D, anti-C, anti-Cw, anti-c, anti-E and anti-e monoclonal antibodies (Moabs) (Hematolog, Russia). Hemagglutination typing in gel columns ID-Cards “DiaClon ABO/D+Reverse Grouping” and “DiaClon Rh-subgroups+K” (Bio-Rad, Switzerland) were used for confirmation of mixed chimerism in transfused patients: agglutinated cells forming a red line on the surface of the gel and no agglutinated cells formed compact button on the bottom of the microtube. DNA extraction was followed by commercial PCR-SSP kits with primers for typing ABO, RHD, and RHCE (BAG, Germany).
Results
We serotyped blood samples from 2055 Russians hematological patients (HP) in 2015. The problems in blood group (BG) identification for ABO system were caused by posttransfused chimerism (PTCh) in 32 primary patients (1,6%) and for Rhesus system – in 64 primary patients (3,1%). 24 patients provided informed consent for genotyping. All patients suffered with idiopathic or symptomatic cytopenia or separate anemia (aplastic anemia -5, different forms of acute leukemia -3, myelodisplastic syndrome -4, multiple myeloma – 2, β-thalassemia -1, PNH – 1, unidentified anemia – 6, partial red cell aplasia – 1, with renal transplant -1) and were transfused before hospitalization to National Research Centre for Hematology. Two patients had a problem with identification in ABO BG: 50% chimerism was detected with anti-A and anti-B Moabs in one and 50% no agglutinated RBC with anti-A – in another one. Genotyping revealed B1B1 and A1B1 BG in these patients. We couldn’t identified Rhesus phenotype in 23 HP: RhC – in 17 (percent of agglutinated RBC ranged from 40 to 90), Rhc – in 13 (agglutinated RBC 50%>90%), RhD – in 4 (agglutinated RBC 20%>90%), RhE – in 18 (agglutinated RBC 30%>90%), Rhe – in 7 (agglutinated RBC 50%>95%), RhCw – in 3 (agglutinated RBC 5%>50%). So PTCh for 1 Rh antigen was detected in 5 HP, for 2 Rh antigens – in 7, for 3 – in 6, for 4 – in 5 and for 5 Rh antigens – in one. The presence of RhCw was confirmed in two HP with 50% RhCw chimerism by genotyping. Molecular method predicted the presence of RhC in 5 HP with chimerism from 20% to 95%, Rhc – in 9 HP (chimerism ranged from 40% to 90%), RhD – in 3 (chimerism ranged from 20% to 90%), RhE – in 9 (chimerism ranged from 30% to 90%) and Rhe – in 3 HP with chimerism 95%. Monitorig of BG in all HP after four month
Conclusion
The determination of ABO and Rhesus phenotypes is very important for previously transfused patients and for patients with known irregular antibodies. The erythrocyte’s genomics has enabled the application of molecular methods to transfusion medicine and allows preventing the alloimmunization to the most clinically significant blood group antigens.
Session topic: E-poster
Keyword(s): Anemia, Chimerism, Transfusion
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