视神经脊髓炎谱系疾病外周血IgH-CDR3免疫组库多样性及其与疾病的相关性

A preliminary study on the immune repertoire of peripheral blood IgH-CDR3 in patients with NMOSD

目的初步建立视神经脊髓炎谱系病患者外周血B淋巴细胞受体的重链互补决定区3(IgH-CDR3)的免疫组库,研究其免疫组库多样性和IgH-CDR3优势取用序列与疾病的相关性。方法收集6例NMOSD患者及6例健康志愿者,并根据治疗情况将NMOSD组分为治疗早期组及稳定期组。运用多重PCR技术对研究对象外周血B细胞IgH-CDR3区域的核苷酸序列进行扩增,扩增后的产物采用第二代高通量测序技术进行免疫组库测序,结合临床资料进行分析。采用非参数检验进行统计学分析。结果NMOSD患者组外周血B细胞IgH-CDR3总数、种类及多样性较正常对照组均有所下降(P>0.05);治疗早期IgH-CDR3序列库的多样性较稳定期高(P>0.05);特征性序列“CASSICLGSGCGGYYYGMDVW”在NMOSD治疗早期患者组中的克隆频次明显高于稳定期NMOSD患者及健康志愿者(P<0.05)。结论免疫组库多样性及疾病特征性优势取用序列的克隆扩增情况可有效反映NMOSD患者的疾病状态,并可评估其治疗效果,或可为临床免疫治疗提供相应的分子靶点和重要的理论基础。

Objective An Immune Repertoire of IgH-CDR3 in peripheral blood of patients with NMOSD was established.To explore the correlation between the changes in the diversity of Immune Repertoire and the dominant use of the IgH-CDR3 sequence and the occurrence,progression and therapeutic intervention of NMOSD.Methods Six patients with NMOSD and 6 healthy volunteers were collected,and the NMOSD group was divided into the early treatment group and the stable treatment group according to the treatment conditions.The nucleotide sequence of the IgH-CDR3 region of B cells in peripheral blood was specifically amplified by multiple PCR,and the second generation of high-throughput sequencing technology was used to sequence the Immune Repertoire.The sequencing results and clinical data were combined to analyze the Immune Repertoire characteristics of NMOSD patients.The Mann-Whitney U test was used for statistical analysis.Results The total number,specie and diversity of IgH-CDR3 in peripheral blood of NMOSD patients decreased compared with that of healthy volunteers(P>0.05).The diversity of IgH-CDR3 sequence in the early NMOSD patients was generally higher than that in the stable patients(P>0.05).The characteristic sequence"CASSICLGSGCGGYYYGMDVW"was highly expressed in the early NMOSD treatment group,and its clonal frequency was significantly higher than that of the stable NMOSD patients and healthy volunteers(P<0.05).Conclusion The clonal amplification of disease characteristic advantage extraction sequence and the diversity of Immune Repertoire can effectively reflect the disease status of NMOSD patients and evaluate the therapeutic effect.It may provide corresponding molecular targets and important theoretical basis for clinical immunotherapy.