Short-term assessment of BCR repertoires of SLE patients after high dose glucocorticoid therapy with high-throughput sequencing.
Short-term assessment of BCR repertoires of SLE patients after high dose glucocorticoid therapy with high-throughput sequencing.
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DOI：
10.1186/s40064-016-1709-4
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Yao X
中科院分区：
文献类型：
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作者：
Shi B;Yu J;Ma L;Ma Q;Liu C;Sun S;Ma R;Yao X
我们对SLE患者在大剂量糖皮质激素治疗前后的BCR谱进行分析和评估，以解决两个基本问题：（1）治疗后，SLE患者的BCR谱在克隆水平上如何变化？ (2) 如何从SLE患者的BCR库中筛选假定的自身抗体克隆集？收集两名SLE患者（P1和P2）不同时间点的PBMC，并提取这些样本的DNA。高通量测序技术应用于BCR库的检测。最后，我们使用生物信息学方法来分析序列数据。我们发现这两名患者在治疗后与治疗前相比失去了一些IGHV3家族基因的使用。对于 IGHV-IGHJ 基因配对，每位患者均未显示出显着变化。此外，对H-CDR3的组成分析显示，每个SLE患者三个时间点的H-CDR3的整体AA组成非常相似，并且具有相同长度（14个AA）和相同位置的H-CDR3 AA使用结果相似。 SLE患者抗核抗体检测显示治疗后部分抗核抗体水平降低；然而，没有迹象表明 BCR 库中自身抗体克隆的百分比会减少。短期大剂量糖皮质激素治疗对SLE患者的BCR库组成影响不大。治疗可以减少蛋白质水平上的自身抗体量，但可能不会减少克隆水平上的 BCR 库中自身抗体克隆的百分比。本文的在线版本 (doi:10.1186/s40064-016-1709-4) 包含补充材料，可供授权用户使用。
We analyze and assess BCR repertoires of SLE patients before and after high dose glucocorticoid therapy to address two fundamental questions: (1) After the treatment, how the BCR repertoire of SLE patient change on the clone level? (2) How to screen putative autoantibody clone set from BCR repertoire of SLE patients? The PBMCs of two SLE patients (P1 and P2) at different time points were collected, and DNA of these samples were extracted. High-throughput sequencing technology was applied in detection of BCR repertoire. Finally, we used bioinformatic methodology to analyse sequence data. We found that these two patients lost some IGHV3 family genes usage after treatment compared with before treatment. For pairing of IGHV–IGHJ gene, no significant change was shown for each patient. In addition, analyses of the composition of H-CDR3 showed overall AA compositions of H-CDR3 at three time points in each SLE patients were very similar, and the results of H-CDR3 AA usage that had the same length (14 AA) and the same position were similar. Antinuclear antibody tests of SLE patients showed that level of some antinuclear antibodies reduced after treatment; however, there was no sign that the percentage of autoantibody clones in BCR repertoires would reduce. High dose glucocorticoid treatment in short term will have little impact on composition of BCR repertoire of SLE patient. Treatment can reduce the amount of autoantibody in the protein level, but may not reduce the percentage of autoantibody clones in BCR repertoire in the clonal level. The online version of this article (doi:10.1186/s40064-016-1709-4) contains supplementary material, which is available to authorized users.