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

Clinical and Experimental Immunology
Serge E DohmenC E van der Schoot

Abstract

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 fr...Continue Reading

References

Dec 1, 1988·International Journal of Bio-medical Computing·L W StrijboschW A Buurman
Sep 1, 1994·British Journal of Haematology·N C Hughes-JonesW H Ouwehand
May 8, 2001·Experimental and Clinical Immunogenetics·M P Lefranc
Jun 22, 2001·The Journal of Immunology : Official Journal of the American Association of Immunologists·V RacanelliF Dammacco

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