Ye Qing, Wang Dong-Jie, Lan Bing, Mao Jian-Hua
Department of Clinical Laboratory, Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, National Children's Regional Medical Center, Hangzhou, 310052, China.
Department of Nephrology, Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, National Children's Regional Medical Center, Hangzhou, 310052, China.
World J Pediatr. 2023 Mar;19(3):273-282. doi: 10.1007/s12519-022-00640-3. Epub 2022 Nov 30.
Clinical studies suggest that the dysfunction of T cells and B cells may play an essential role in the pathogenesis of idiopathic nephrotic syndrome (INS), but laboratory evidence is lacking. Therefore, this study explored T-cell receptor (TCR) and B-cell receptor (BCR) profiling in children with idiopathic nephrotic syndrome.
High-throughput sequencing technology was used to profile the TCR and BCR repertoires in children with INS. Peripheral blood was collected from ten INS patients, including five vinculin autoantibody-positive patients and five vinculin autoantibody-negative patients, before and after treatment. TCR and BCR libraries were constructed by 5'-RACE and sequenced by a DNBSEQ-T7 sequencer, and sequence analyses were performed using ReSeqTools, FastP, MiXCR, and VDJtools.
The TRA (T-cell receptor α), TRG (T-cell receptor γ), and IGH (immunoglobulin heavy chain) repertoires of the INS group were occupied by highly abundant clonotypes, whereas small clonotypes occupied the healthy group, especially TRA. A significant increase in the Shannon-Weaver index was observed for the TRA and TRG repertoires after treatment in vinculin autoantibody-negative patients, but a significant increase in the IGH repertoire after treatment was observed in vinculin autoantibody-positive patients. The frequency of some V-J pairs was significantly enriched in steroid-sensitive nephrotic syndrome patients. The usage frequency of the V and J genes was skewed in patients, which seemed not related to immunosuppressive therapy. However, after effective treatment, dynamic changes in the size of the individual clonotype were observed.
T-cell and B-cell immunity contribute to the pathogenesis of different INSs. Video: (MP4 99,786 KB).
临床研究表明，T细胞和B细胞功能障碍可能在特发性肾病综合征（INS）的发病机制中起重要作用，但缺乏实验室证据。因此，本研究探讨了特发性肾病综合征患儿的T细胞受体（TCR）和B细胞受体（BCR）谱。
采用高通量测序技术分析INS患儿的TCR和BCR库。收集10例INS患者（包括5例纽蛋白自身抗体阳性患者和5例纽蛋白自身抗体阴性患者）治疗前后的外周血。通过5'-RACE构建TCR和BCR文库，并用DNBSEQ-T7测序仪进行测序，使用ReSeqTools、FastP、MiXCR和VDJtools进行序列分析。
INS组的TRA（T细胞受体α）、TRG（T细胞受体γ）和IGH（免疫球蛋白重链）库由高度丰富的克隆型占据，而健康组则由小克隆型占据，尤其是TRA。纽蛋白自身抗体阴性患者治疗后TRA和TRG库的香农-韦弗指数显著增加，而纽蛋白自身抗体阳性患者治疗后IGH库显著增加。一些V-J对的频率在类固醇敏感型肾病综合征患者中显著富集。患者中V和J基因的使用频率存在偏差，这似乎与免疫抑制治疗无关。然而，有效治疗后，观察到个体克隆型大小的动态变化。
T细胞和B细胞免疫在不同类型的INS发病机制中起作用。视频：（MP4 99,786 KB）
