PUBLICATION

Transgenic overexpression of BAFF regulates the expression of immune-related genes in zebrafish, Danio rerio

Authors
Zhang, L., Liu, C., Zhou, X., Xie, Y., Su, L., Geng, Q., Liu, B., Liu, S.
ID
ZDB-PUB-161221-6
Date
2016
Source
Journal of genetics   95: 751-760 (Journal)
Registered Authors
Keywords
none
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • B-Cell Activating Factor/genetics*
  • Cell Line
  • Cloning, Molecular
  • Gene Expression*
  • Gene Expression Regulation
  • Gene Order
  • Genes, Reporter
  • Genetic Vectors/genetics
  • Humans
  • Immunity/genetics*
  • Transfection
  • Zebrafish/genetics*
  • Zebrafish/immunology*
PubMed
27994173 Full text @ J. Genet.
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.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping