Kotowski M, Bogacz A, Bartkowiak-Wieczorek J, Bukowska A, Surowiec N, Dziewanowski K, Czerny B, Grześkowiak E, Ostrowski M, Machaliński B, Sieńko J
Department of General Pathology, Pomeranian Medical University, Szczecin, Poland; Department of General Surgery and Transplantation, Pomeranian Medical University, Szczecin, Poland.
Department of Stem Cells and Regenerative Medicine, Institute of Natural Fibres and Medicinal Plants, Poznań, Poland; Department of Histocompatibility with Laboratory of Genetic Diagnostics, Regional Blood Centre, Poznan, Poland.
Transplant Proc. 2018 Jul-Aug;50(6):1605-1615. doi: 10.1016/j.transproceed.2018.05.005. Epub 2018 Jun 21.
Thanks to new generation sequencing (NGS) and expansion of HLA typing with additional loci, it will be possible to increase the effectiveness of graft survival and to avoid complications related to the immune system. New pharmacogenetic factors are still being researched to develop better immunosuppressive treatment.
The incidence of polymorphic HLA loci variants was established, based on a high-resolution NGS method in kidney graft recipients. Furthermore, haplotypic analysis between examined loci was conducted to type additional loci that may influence the transplantation result. A total of 120 kidney recipients were enrolled in the study. A commercial DNA extraction kit in Tubes (QIAamp DNA Blood Mini Kit Qiagen, Germany) was used to isolate DNA from the blood. Sequencing library preparation was done with TruSight HLA set. The Conexio computer program was used to analyse the results of HLA typing.
The patients with alleles A02:01:01, B44:02:01, C03:03:01, C01:02:01, C05:01:01, C07:02:01, DQB103:03:02, DQB106:04:01, or with haplotypic variation A25:01:01-B18:01:01- C15:01:01 were taking the highest doses of cyclosporine (CsA), in contrast to patients with allele B18:01:01, DQB106:02:01, DQB102:02:01, or haplotypic variation A02:01:01- B44:02:01-C01:01:01, who were taking the lowest doses. The highest dose of tacrolimus (TAC) was administered to patients with alleles A68:01:02, A29:01:01, B07:02:01, B35:02:01, B38:01:01, DRB112:01:01, DQB105:03:01, or haplotypic variations A02:01:01-B57:01:01-C07:01:01, A03:01:01-B07:02:01-C13:01:01, A29:02:01-B44:03:01- C07:01:01, and A01:01:01-B08:01:01-C03:01:01. Additionally, it was established that HLA-DRB3, HLA-DRB4, HLA-DRB5, HLA-DPA1, and HLA-DQA1 show very slight polymorphism, which suggests that there is no need for their typing for transplantation purposes. Moreover, loci HLA-C, HLA-DQB1, and HLA-DPB1, which are not routinely examined in recipient-donor matching, show genetic variability that may increase the risk of transplant rejection or shortened graft life.
Expanding the qualification procedure to include allele genotyping could allow clinicians to establish immunosuppressive treatment schemes that would be optimally suited for recipients' phenotype.
得益于新一代测序技术（NGS）以及增加位点进行的HLA分型扩展，提高移植物存活的有效性并避免与免疫系统相关的并发症将成为可能。目前仍在研究新的药物遗传学因素以开发更好的免疫抑制治疗方法。
基于高分辨率NGS方法确定肾移植受者中多态性HLA位点变体的发生率。此外，对检测位点之间进行单倍型分析，以确定可能影响移植结果的其他位点。共有120名肾移植受者纳入本研究。使用商业化的Tubes DNA提取试剂盒（德国Qiagen公司的QIAamp DNA Blood Mini Kit）从血液中分离DNA。采用TruSight HLA试剂盒进行测序文库制备。使用Conexio计算机程序分析HLA分型结果。
携带等位基因A02:01:01、B44:02:01、C03:03:01、C01:02:01、C05:01:01、C07:02:01、DQB103:03:02、DQB106:04:01，或单倍型变异A25:01:01 - B18:01:01 - C15:01:01的患者服用环孢素（CsA）的剂量最高，而携带等位基因B18:01:01、DQB106:02:01、DQB102:02:01，或单倍型变异A02:01:01 - B44:02:01 - C01:01:01的患者服用剂量最低。携带等位基因A68:01:02、A29:01:01、B07:02:01、B35:02:01、B38:01:01、DRB112:01:01、DQB105:03:01，或单倍型变异A02:01:01 - B57:01:01 - C07:01:01、A03:01:01 - B07:02:01 - C13:01:01、A29:02:01 - B44:03:01 - C07:01:01以及A01:01:01 - B08:01:01 - C03:01:01的患者服用他克莫司（TAC）的剂量最高。此外，已确定HLA - DRB3、HLA - DRB4、HLA - DRB5、HLA - DPA1和HLA - DQA1显示出非常轻微的多态性，这表明就移植目的而言无需对其进行分型。而且，在供受者配型中通常不进行检测的HLA - C、HLA - DQB1和HLA - DPB1位点显示出遗传变异性，这可能会增加移植排斥风险或缩短移植物存活时间。
将基因分型纳入配型程序可使临床医生制定出最适合受者表型的免疫抑制治疗方案。
