用长读序列测序在转录组水平上进行全长免疫谱的重建和分析
Xuanmei Luo1,2, Lili Zhang1, Yifei Li1
1Clinical Biobank, Beijing Hospital, National Center of Gerontology, National Health Commission, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing, China.
Clinical chemistry
|September 17, 2024
概括
全长免疫谱系测序 (FLIRseq) 为分析V(D) J重组和免疫球蛋白类提供了一种卓越的方法. 这种先进的技术有助于理解疾病机制和个性化细胞疗法.
科学领域:
- 免疫学 免疫学 免疫学
- 基因组学就是基因组学.
- 生物信息学是一种生物信息学.
背景情况:
- 传统的短读测序与多样化的免疫谱 (IR) 斗争.
- 重建全长的IR对于理解免疫反应和疾病至关重要.
研究的目的:
- 开发和验证全长免疫谱系测序 (FLIRseq) 的新的工作流程.
- 与现有方法相比,评估FLIRseq的准确性,量化和全面性.
- 为了分析各种患者队伍的免疫谱.
主要方法:
- 使用线性滚动圆放大和纳米孔测序开发了FLIRseq工作流.
- 对等离子体混合物和短读BCR/TCR-seq.进行验证的准确性和量化.
- 应用FLIRseq分析白血病,过敏性疾病,牛皮和前列腺癌患者的IR.
主要成果:
- FLIRseq显示了比标准的纳米孔测序更高的准确性 (较低的不匹配/差距率,更高的身份率).
- FLIRseq提供了更全面的数据,包括CDR3,同型和罕见的V基因,超过了BCR/TCR-seq.
- 观察到克隆型多样性的显著变化以及白血病缓解中的异常BCR/TCR百分比.
- 在过敏性疾病和牛皮中获得了V(D) J重组和免疫球蛋白类的洞察力.
- 在牛皮组织中确定了对T细胞受体β链的一致AlphaFold2预测蛋白质结构.
结论:
- FLIRseq使V(D) J重组和免疫球蛋白类的公正,全面的分析成为可能.
- FLIRseq有助于表征致病机制和监测最小残留疾病.
- FLIRseq支持采用细胞疗法的定制.
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