The analysis and quantification of a clonal B-cell response in a hyperimmunized anti-D donor

S. E. Dohmen, A. Verhagen, S. M. De Groot, Lisa Marie Stott, Stanislaw Urbaniak, C. E. Van Der Schoot

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)


Healthy volunteers are hyperimmunized with RhD-positive red cells in order to obtain plasma containing high titres of anti-D immunoglobulin, which is used for the prevention of haemolytic disease of the fetus and newborn. We analysed the anti-D immune response in a donor who had been hyperimmunized for 7 years and who showed declining anti-D titres despite re-immunization. A phage display library representing the complete immunorepertoire and a second library representing the IGHV3 superspecies family genes (IGHV3s) repertoire in the donor were constructed and analysed. A clonal Ig-gene rearrangement was quantified in the peripheral blood by limiting dilution polymerase chain reaction (PCR) All RhD-binding phages from both libraries, except one, had heavy chains with IGH-VDJ rearrangements of the same clonal origin, but with different patterns of somatic mutations and joined with different light chains. Limiting dilution PCR performed on mRNA and genomic DNA showed a frequency of 1 clonal B cell in 2000 IgG1/3-positive B cells. We show the presence of clonally related RhD-specific B cells in a hyperimmunized anti-D donor who had declining anti-D titres and who was unresponsive to re-immunization. Furthermore, we found a high frequency of clonal B cells. These results contribute to the understanding of the immune response against RhD in hyperimmunized anti-D donors.

Original languageEnglish
Pages (from-to)223-232
Number of pages9
JournalClinical and Experimental Immunology
Issue number2
Early online date27 Mar 2006
Publication statusPublished - May 2006


  • anti-D
  • clonal B cells
  • hyperimmunization
  • phage display
  • phage-display library
  • D antibodies
  • anti-RH(D) antibodies
  • gene segments
  • quantitation
  • specificity
  • mechanism
  • usage


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