The RHCE allele ceRT: D epitope 6 expression does not require D-specific amino acids.

BACKGROUND: False-positive D typing in patients may lead to anti-D immunization caused by D+ transfusions or by omission of anti-D prophylaxis. Known causes of such errors are RhCE variants carrying RhD-specific amino acids and cold agglutinin activity of some frequently used monoclonal anti-D. STUDY DESIGN AND METHODS: The molecular basis of eight samples referred because of "false-positive" reactions with some commercial monoclonal anti-D was investigated by PCR and nucleotide sequencing from genomic DNA. PCR with sequence-specific priming was developed to specifically detect the underlying aberrant RHCE allele. The D epitope profile of the allele was determined by serology. RESULTS: The aberrant reactivity of the samples was caused by the RHCE allele RHCE(R154T) that occurred in a cde haplotype. The phenotype dubbed ceRT expressed the important D epitope 6, which is the target epitope of most monoclonal anti-D used in routine typing. DISCUSSION: The characterization of ceRT demonstrated a previously unknown mechanism of antigen D expression that does not require any D-specific amino acid. At least for some D epitopes, D-like structures may be mimicked by RhCE proteins carrying amino acid substitutions not representative for RhD.

The DAU allele cluster of the RHD gene.

Variant D occurs frequently in Africans. However, considerably less RHD alleles have been described in this population compared with Europeans. We characterized 5 new RHD alleles, dubbed DAU-0 to DAU-4, that shared a T379M substitution and occurred in a cDe haplotype. DAU-1 to DAU-4 were detected in Africans with partial D phenotypes. They harbored one and 2 additional missense mutations, respectively, dispersed throughout the RhD protein. An anti-D immunization was found in DAU-3. DAU-0 carrying T379M only was detected by screening European blood donors and expressed a normal D phenotype. Within the phylogeny of the RHD alleles, DAU formed an independent allele cluster, separate from the DIVa, weak D type 4, and Eurasian D clusters. The characterization of the RH phylogeny provided a framework for future studies on RH alleles. The identification of the DAU alleles increased the number of known partial D alleles in Africans considerably. DAU alleles may be a major cause of antigen D variability and anti-D immunization in patients of African descent.