Original article

Blood Transfusion 2-2018 (March - April)

Single-stranded DNA aptamer targeting and neutralization of anti-D alloantibody: a potential therapeutic strategy for haemolytic diseases caused by Rhesus alloantibody

Authors

Key words: single-stranded DNA, aptamer, RhD alloantibody, delayed haemolytic transfusion reactions, haemolytic disease of the newborn
Publication Date: 2016-11-11

Abstract

Background. Rhesus (Rh) D antigen is the most important antigen in the Rh blood group system because of its strong immunogenicity. When RhD-negative individuals are exposed to RhD-positive blood, they may produce anti-D alloantibody, potentially resulting in delayed haemolytic transfusion reactions and Rh haemolytic disease of the foetus and newborn, which are difficult to treat. Inhibition of the binding of anti-D antibody with RhD antigens on the surface of red blood cells may effectively prevent immune haemolytic diseases.
Materials and methods. In this study, single-stranded (ss) DNA aptamers, specifically binding to anti-D antibodies, were selected via systematic evolution of ligands by exponential enrichment (SELEX) technology. After 14 rounds of selection, the purified ssDNA was sequenced using a Personal Genome Machine system. Haemagglutination inhibition assays were performed to screen aptamers for biological activity in terms of blocking antigen-antibody reactions: the affinity and specificity of the aptamers were also determined.
Results. In addition to high specificity, the aptamers which were selected showed high affinity for anti-D antibodies with dissociation constant (Kd) values ranging from 51.46±14.90 to 543.30±92.59 nM. By the combined use of specific ssDNA aptamer 7 and auxiliary ssDNA aptamer 2, anti-D could be effectively neutralised at low concentrations of the aptamers.
Discussion. Our results demonstrate that ssDNA aptamers may be a novel, promising strategy for the treatment of delayed haemolytic transfusion reactions and Rh haemolytic disease of the foetus and newborn.

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Authors

Yinze Zhang - Shenzhen Blood Centre, Institute of Transfusion Medicine, Shenzhen

Fan Wu - Shenzhen Blood Centre, Institute of Transfusion Medicine, Shenzhen

Manni Wang - Shaanxi Blood Centre, Xi'an

Naibao Zhuang - Shenzhen Blood Centre, Institute of Transfusion Medicine, Shenzhen

Huayou Zhou - Nanfang Hospital, Southern Medical University, Guangzhou, China

Hua Xu - Shaanxi Blood Centre, Xi'an

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