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The Impact of Using Genotyped Reagent Red Blood Cells in Antibody Identification.

ABSTRACT: Background:The detection and identification of antibodies to red blood cell (RBC) antigens is one of the most important and challenging issues in transfusion medicine. Up to date there are 354 RBC antigens recognized by the International Society of Blood Transfusion (ISBT). The reagent RBCs used in commercial antibody screening and identification panels however are usually serologically typed for up to 40 clinically important antigens. Thus the identification of many antibody specificities remains impossible when using reagent RBCs with only limited information about their antigens. To improve the pre-transfusion diagnostics, we developed antibody identification panels with reagent RBCs serologically typed for 26 antigens and additionally genotyped for 30 blood group alleles. Methods:The reagent RBCs in the panels were characterized serologically for the clinically most significant 'standard' antigens. The reagent RBC donors were additionally genotyped by using in-house PCR-SSP methods. The antibody identification was performed in the indirect antiglobulin test using untreated and papain-treated RBCs in the gel technique. Antibodies identified due to the genotype information were confirmed by serology using appropriate reference RBCs. Results:Within a time period of 3 years and 8 months, 16,878 blood samples from 8,467 patients were tested in our reference laboratory. In total, 21 different antibodies from 10 different blood group systems could be identified in 126 patients (1.5%) due to the genotype information obtained for the reagent RBCs. Antibodies to antigens from the Knops system (53 patients; 42%, 8 patients with anti-Knb) and to Cartwright antigens (31 patients; 25%) were the most frequent. Conclusion:The use of genotyped reagent RBCs in antibody identification panels extends the range of detectable antibody specificities, accelerates the antibody identification, and improves the pre-transfusion diagnostics.

SUBMITTER: Scharberg E

PROVIDER: S-EPMC6158584 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature

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The Impact of Using Genotyped Reagent Red Blood Cells in Antibody Identification.

Scharberg Erwin E Rink Gabi G Portegys Jan J Rothenberger Sina S Gillhuber Nicole N Richter Ekkehard E Bugert Peter P

Transfusion medicine and hemotherapy : offizielles Organ der Deutschen Gesellschaft fur Transfusionsmedizin und Immunhamatologie 20180724 4

<h4>Background</h4>The detection and identification of antibodies to red blood cell (RBC) antigens is one of the most important and challenging issues in transfusion medicine. Up to date there are 354 RBC antigens recognized by the International Society of Blood Transfusion (ISBT). The reagent RBCs used in commercial antibody screening and identification panels however are usually serologically typed for up to 40 clinically important antigens. Thus the identification of many antibody specificiti ...[more]

PMID: 30283270

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Project description:BackgroundNumerous studies have demonstrated that DNA methylation levels in the aryl hydrocarbon receptor repressor (AHRR) gene measured in cord blood are significantly associated with prenatal tobacco smoke exposure and can be used as a fetal exposure biomarker. The mechanism driving this demethylation has not been determined and it is unclear if all cord blood cell types are impacted. Nucleated red blood cells (nRBCs/CD235a+ cells) are developmentally immature RBCs that display genome-wide hypomethylation and are observed at increased frequency in the cord blood of smoking mothers. We tested if AHRR methylation levels in CD235a+ nRBCs or nRBC counts influenced AHRR methylation in whole cord blood.MethodsCord blood was collected from smoking (n = 34) and nonsmoking (n = 19) mothers and DNA was prepared from whole cord blood, isolated CD235a+ nRBCs, and CD14+ monocytes. AHRR methylation in cord blood DNA was measured using Illumina 850K arrays (cg05575921, chr5:373378). Pyrosequencing was used to compare methylation levels among cord blood, CD235a+, and CD14+ cells. We measured nRBC percentages using conventional complete blood counts and estimated percent nRBCs by a deconvolution model.ResultsMethylation levels in AHRR were significantly lower in nRBCs relative to whole cord blood and CD14+ monocytes. While AHRR methylation levels in the cell types were significantly correlated across all subjects, methylation values at the chr5:373378 CpG averaged 14.6% lower in nRBCs (range 0.4 to 24.8%; p = 3.8E-13) relative to CD14+, with nonsmokers showing a significantly greater hypomethylation (- 4.1%, p = 1.8E-02). Methylation level at the AHRR chr5:373378 CpG was strongly associated with self-reported smoking in both CD14+ monocytes (t test p = 5.7E-09) and nRBCs (p = 4.8E-08), as well as cotinine levels (regression p = 1.1E-07 and p = 3.6E-04, respectively). For subjects with whole blood 850K data, robust linear regression models adjusting for estimated cell type composition, either including nRBCs counts or estimates, modestly increased the association between smoking and cg05575921 methylation.ConclusionsPrenatal smoke exposure was highly significantly associated with AHRR methylation in cord blood, CD14+ monocytes, and CD235a+ nRBCs. AHRR methylation levels in nRBCs and nRBC counts had minimal effect on cord blood methylation measurements. However, regression models using estimated nRBCs or actual nRBC counts outperformed those lacking these covariates.

Dataset Information

The Impact of Using Genotyped Reagent Red Blood Cells in Antibody Identification.

Publications

The Impact of Using Genotyped Reagent Red Blood Cells in Antibody Identification.

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