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scBCR-seq revealed a special and novel IG H&L V(D)J allelic inclusion rearrangement and the high proportion dual BCR expressing B cells

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Abstract

Since the initial report of V (D) J “allelic exclusion/inclusion” (allelic exclusion rearrangement or allelic inclusion rearrangement) and the concept of the “dual B cell receptor (BCR)” in 1961, despite ongoing discoveries, the precise proportion and source mechanism of dual BCR under physiological conditions have been puzzling immuologists. This study takes advantage of the single cell B cell receptor sequencing (scBCR-seq) technology, which can perfectly match the heavy and light chains of BCR at the level of a single B cell, and obtain the full length mRNA sequence of the complementary determining region 3 (CDR3). Through analyzing the pairing of functional IGH (immunoglobulin heavy chain) and IGL (immunoglobulin light chain) in single B cell from both human and mouse bone marrow and peripheral blood, it was observed that dual BCR B cells exhibit stable and high levels of expression. Among them, the human bone marrow and peripheral blood contain about 10% dual (or multiple) BCR B cells, while in mouse peripheral blood and bone marrow memory B cells, this proportion reaches around 20%. At the same time, we innovatively found that in each research sample of humans and mice, there are three (or more) functional rearrangements (mRNA level) of a single chain in a single B cell. By analyzing the position, direction and other compositional characteristics of the V(D)J gene family, we found that at least two (or more) of them are derived from over two (or more) specific allelic inclusion rearrangements of a single chromosome (mRNA molecular level evidence), our findings also highlighted the necessity of classified single cell sequencing data based on single, dual (or multiple) and cannot be assembled into BCR when analyzing the B cell repertoire. The results of this article provides new methods and modeling references for evaluating the proportion and source mechanisms of dual BCR B cells, as well as potential significance of allelic inclusion (exclusion escape) of V(D)J rearrangement.

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Acknowledgements

We would like to express our gratitude to the 10×Company, GEO and IMGT databases for providing the availability of the data. In addition, thanks to Shi, Z, Peterson, J.N, Wang P and Riedel R for sharing the single cell sequencing data.

Funding

This study was supported by the National Natural Science Foundation of China (82160279) and the Guizhou Provincial Hundred level Talent Fund [No. (2018)5637].

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  1. Lanwei Zhu, Qi Peng, and Yingjie Wu are joint first authors.

Authors and Affiliations

  1. Department of Immunology, Center of Immunomolecular Engineering, Innovation and Practice Base for Graduate Students Education, Zunyi Medical University, Zunyi, China

    Lanwei Zhu, Qi Peng, Yingjie Wu & Xinsheng Yao

Authors
  1. Lanwei Zhu
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  2. Qi Peng
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  3. Yingjie Wu
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  4. Xinsheng Yao
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Contributions

XY completed the experimental design and paper writing, LZ, QP, and YW completed the data analysis and chart making.

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Correspondence to Xinsheng Yao.

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Zhu, L., Peng, Q., Wu, Y. et al. scBCR-seq revealed a special and novel IG H&L V(D)J allelic inclusion rearrangement and the high proportion dual BCR expressing B cells. Cell. Mol. Life Sci. 80, 319 (2023). https://doi.org/10.1007/s00018-023-04973-8

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