Van de Velde H, Georgiou I, De Rycke M, Schots R, Sermon K, Lissens W, Devroey P, Van Steirteghem A, Liebaers I
Centre for Reproductive Medicine, Brussels, Belgium.
Hum Reprod. 2004 Mar;19(3):700-8. doi: 10.1093/humrep/deh153. Epub 2004 Jan 29.
Beta-Thalassaemia results from co-inheritance of two mutant beta-globin alleles. Allogeneic cord blood cell transplantation (CBT) from an HLA-identical sibling donor is an excellent treatment option for beta-thalassaemia. In families with an affected child and willing to have another child, IVF followed by preimplantation genetic diagnosis (PGD) can be applied to exclude affected embryos. Furthermore, healthy embryos could be HLA matched with the affected child so that cord blood from the future newborn can be used to transplant the affected sibling.
We developed an indirect single-cell HLA typing technique based on the use of a bank of seven microsatellite markers within the HLA locus from which four informative and evenly distributed markers were selected.
The methodology was validated in three beta-thalassaemia families having six ovarian stimulation cycles in view of IVF and PGD. Six PGD cycles were performed in two families. On 58 embryos tested, the combined PCR was successful in 54 (93%). Two transfers were done and one clinical pregnancy was obtained. Using confirmatory analysis on 50 embryos, the accuracy for HLA typing was 100%.
This strategy offers a new therapeutic option for patients with beta-thalassaemia and other monogenic diseases that can be cured with CBT.
β地中海贫血是由两个突变的β珠蛋白等位基因共同遗传导致的。来自 HLA 相同的同胞供体的异基因脐血细胞移植（CBT）是β地中海贫血的一种极佳治疗选择。在有患病儿童且愿意再生育一个孩子的家庭中，体外受精（IVF）后进行植入前基因诊断（PGD）可用于排除受影响的胚胎。此外，健康胚胎可与患病儿童进行 HLA 配型，以便未来新生儿的脐血可用于移植给患病的同胞。
我们基于使用一组位于 HLA 基因座内的七个微卫星标记开发了一种间接单细胞 HLA 分型技术，从中选择了四个信息丰富且分布均匀的标记。
鉴于 IVF 和 PGD，该方法在三个有六个卵巢刺激周期的β地中海贫血家庭中得到验证。在两个家庭中进行了六个 PGD 周期。在检测的 58 个胚胎中，联合 PCR 在 54 个（93%）中成功。进行了两次移植，获得了一次临床妊娠。对 50 个胚胎进行确证分析，HLA 分型的准确率为 100%。
该策略为β地中海贫血患者以及其他可通过 CBT 治愈的单基因疾病患者提供了一种新的治疗选择。
