Abstract
The B-cell activating factor (BAFF) is a member of tumour necrosis factor (TNF) superfamily that specifically regulates B lymphocyte proliferation and survival. Excess BAFF leads to overproduction of antibodies for secretion, anti-dsDNA antibodies and a lupus-like syndrome in mice. To investigate whether transgenic overexpression of the zebrafish BAFF leads to immunoglobulin changes and/or early maturing of the immune system, a Tol2-GFP-2A-BAFF/His recombinant plasmid was constructed by inserting a 2A peptide between the green fluorescent protein (GFP) and BAFF sequences. Functional GFP and BAFF proteins were expressed separately and confirmed in HeLa cells. The relative expression of immune-related genes (IgLC-1, IgLC-2, IgLC-3, IgD, IgM and IL-4), early lymphoid markers (Ikaros, Rag-1 and TCRAC), and the protooncogene Bcl-2 were evaluated by quantitative polymerase chain reaction (PCR) in F0 founder of transgenic zebrafish juveniles and adults. Ectopic expression of BAFF in adults was confirmed using Western blots and was shown to upregulate IgLC-1, IgLC-2, IgD, IgM, IgZ/T, Ikaros, Rag-1, TCRAC, IL-4 and Bcl-2 expression in juveniles on day 21 and IgLC-1, IgLC-2, IgD, IgM, IgZ/T, Rag-1, TCRAC and Bcl-2 expression in zebrafish three months postfertilization. The relative titers of specific IgM against Edwardsiella tarda WED were assessed using modified enzyme-linked immunosorbent assay (ELISA) with the whole body homogenate of zebrafish and demonstrated a significant increase in BAFF-transgenic group. Therefore, our findings provided novel insight into further exploration of modulating adaptive immunity and studying autoimmune diseases caused by regulating BAFF.
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Acknowledgements
This work was supported by the Construction Programme of Hebei Scientific Conditions (13967607D, 12965519D and 12966121D) and National Natural Science Foundation of Hebei province (C2012206096).
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Corresponding editor: Indrajit Nanda
[Zhang L., Liu C., Zhou X., Xie Y., Su L., Geng Q., Liu B. and Liu S. 2016 Transgenic overexpression of BAFF regulates the expression of immune-related genes in zebrafish, Danio rerio. J. Genet. 95, xx–xx]
Li Zhang and Chao Liu contributed equally to this work.
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ZHANG, L., LIU, C., ZHOU, X. et al. Transgenic overexpression of BAFF regulates the expression of immune-related genes in zebrafish, Danio rerio . J Genet 95, 751–760 (2016). https://doi.org/10.1007/s12041-016-0690-6
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DOI: https://doi.org/10.1007/s12041-016-0690-6