背景:关于肾移植中B细胞亚型和B细胞受体V(D)J基因使用的研究很少。这项研究旨在调查ABO不相容(ABOi)肾移植(KT)患者与ABO相容(ABOc)KT患者的V(D)J基因片段使用情况。
方法:我们选择了16例适应(ABOiA)的ABOiKT患者,6例ABOc稳定的KT患者(ABOcS),和6例ABOiKT患者在第10天出现活检证实的急性抗体介导的排斥反应(ABOiR),其移植组织样本在2010年至2014年期间储存在生物存储库中.对移植物组织的完整转录组进行测序并通过RNA测序(RNA-seq)进行分析。国际ImMunoGeneTics信息系统(IMGT®)用于V(D)J基因片段使用的深入比较。
结果:28名KT接受者的平均年龄为43.3±12.8岁,53.6%为男性。通过家庭,IGHV3、IGHJ4、IGLV2和IGLJ3基因片段在所有组中最经常使用,三个患者组的使用情况无统计学差异。虽然IGKV3最常用于ABOiA和ABOiR组,IGKV1最常用于ABOcS组。此外,虽然IGKJ1最常用于ABOiA和ABOcS组,IGKJ4在ABOiR组中最经常使用。根据单个基因片段,IGHV4-34和IGHV4-30-2在ABOiR组中比在ABOiA组中更常用,IGHV6-1在ABOcS组比ABOiR组更常用。IGLV7-43在ABOcS组中比在ABOi组中更常用。然而,技术可变性,小样本量,利妥昔单抗或HLA错配的潜在混杂效应是我们研究的局限性.
结论:我们的研究结果表明,RNA-seq转录组分析可以提供关于B细胞受体的V(D)J基因使用以及ABOiKT中调节和免疫反应机制的信息。
BACKGROUND: Studies on B-cell subtypes and V(D)J gene usage of B-cell receptors in kidney transplants are scarce. This study aimed to investigate V(D)J gene segment usage in ABO-incompatible (ABOi) kidney transplant (KT) patients compared to that in ABO-compatible (ABOc) KT patients.
METHODS: We selected 16 ABOi KT patients with accommodation (ABOiA), 6 ABOc stable KT patients (ABOcS), and 6 ABOi KT patients with biopsy-proven acute antibody-mediated rejection (ABOiR) at day 10, whose graft tissue samples had been stored in the biorepository between 2010 and 2014. Complete transcriptomes of graft tissues were sequenced and analyzed through RNA sequencing (RNA-seq). The international ImMunoGeneTics information system (IMGT®) was used for in-depth comparison of V(D)J gene segment usage.
RESULTS: The mean age of the 28 KT recipients was 43.3 ± 12.8 years, and 53.6% were male. By family, IGHV3, IGHJ4, IGLV2, and IGLJ3 gene segments were most frequently used in all groups, and their usage was not statistically different among the three patient groups. While IGKV3 was most frequently used in both the ABOiA and ABOiR groups, IGKV1 was most commonly used in the ABOcS group. In addition, while IGKJ1 was most commonly used in the ABOiA and ABOcS groups, IGKJ4 was most frequently used in the ABOiR group. According to individual gene segments, IGHV4-34 and IGHV4-30-2 were more commonly used in the ABOiR group than in the ABOiA group, and IGHV6-1 was more commonly used in the ABOcS group than in the ABOiR group. IGLV7-43 was more commonly used in the ABOcS group than in the ABOi group. However, technical variability, small sample size, and potential confounding effects of Rituximab or HLA mismatching are limitations of our study.
CONCLUSIONS: Our findings suggest that RNA-seq transcriptomic analyses can provide information on the V(D)J gene usage of B-cell receptors and the mechanisms of accommodation and immune reaction in ABOi KT.