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Springerplus. 2016 Jan 26;5:75. doi: 10.1186/s40064-016-1709-4. eCollection 2016.

Short-term assessment of BCR repertoires of SLE patients after high dose glucocorticoid therapy with high-throughput sequencing.

Shi B#1, Yu J#2, Ma L3, Ma Q4, Liu C5, Sun S6, Ma R3, Yao X3.

Author information

1
Department of Laboratory Medicine, Zunyi Medical University, Zunyi, 563000 China.
2
Cell Engineering Laboratory, The First Affiliated Hospital of ZunYi Medical University, Zunyi, China.
3
Department of Immunology, Research Center for Medicine and Biology, Innovation and Practice Base for Graduate Students Education, Zunyi Medical University, Zunyi, 563000 China.
4
Central Laboratory, Guizhou Aerospace Hospital, Zunyi, China.
5
Department of Nephropathy and Rheumatology, The First Affiliated Hospital of ZunYi Medical University, Zunyi, China.
6
Department of Breast Surgery, The First Affiliated Hospital of ZunYi Medical University, Zunyi, China.
#
Contributed equally

Abstract

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.

KEYWORDS:

BCR repertoire; H-CDR3; High-throughput sequencing; SLE

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