Citation
- Authors: Shi Y. et al.
- Year: 2023
- Journal: Fish Shellfish Immunol 133 108530
- Applications: in vitro / DNA, Poly (I:C) / jetOPTIMUS
- Cell types:
- Name: EPC
- Name: ZF4
- Name: ZFIN
Method
JetOPTIMUS for transfection of DNA was purchased from PolyPlus.
EPC cells were seeded in 12-well plates overnight and transfected with pcDNA3.1-IL-27A-Flag or pcDNA3.1 vector.
The ZFIN cells were transfected with plasmids encoding SVCV-N, SVCV-P, SVCV-M, SVCV-G and SVCV-L, respectively.
ZF4 cells were seeded in 6-well plates overnight and infected with SVCV (A-E) or transfected with poly(I:C) (4 μg)
Abstract
Interleukin (IL) 27 is a member of the IL-12 family and is a heterodimeric cytokine composed of IL-27A and Epstein-Barr virus-induced 3 (EBI3). It plays an important role in regulating inflammation and cancer progression. IL-27A not only functions by dimerizing with EBI3 but also acts alone. Here, we report that IL-27A and EBI3 suppress spring viremia of carp virus (SVCV) replication in zebrafish. Expression analysis reveals that il-27a and ebi3 were significantly upregulated in the ZF4 cells by SVCV and poly(I:C), and in the zebrafish caudal fin (ZFIN) cells overexpressed with SVCV genes. Interestingly, il-27a and ebi3 were not modulated by IFNφ1, indicating that they are not IFN stimulated genes (ISGs). Furthermore, overexpression of IL-27A and EBI3 alone inhibited SVCV replication in the EPC cells, but less potent than co-expression of IL-27A and EBI3. Intriguingly, IL-27A could not induce the expression of irf3, ifn, isg15 and mx1. Taken together, our results demonstrate that IL-27A and EBI3 activate innate antiviral response in an IFN independent manner in zebrafish.