DNA vaccine dual-expressing viral hemorrhagic septicemia virus glycoprotein and C-C motif chemokine ligand 19 induces the expression of immune-related genes in zebrafish (Danio rerio)

J Microbiol. 2022 Oct;60(10):1032-1038. doi: 10.1007/s12275-022-2231-8. Epub 2022 Aug 1.

Abstract

Glycoprotein (G protein)-based DNA vaccines are effective in protecting aquaculture fish from rhabdoviruses but the degree of immune response they elicit depends on plasmid concentration and antigen cassette. Here, we developed a DNA vaccine using the viral hemorrhagic septicemia virus G (VG) gene and chemokine (C-C motif) ligand 19 (CCL19)a.2 regulated by the CMV promoter as the molecular adjuvant. After transfection of the prepared plasmid (pVG + CCL19) into epithelioma papulosum cyprini cells, mRNA expression was confirmed through quantitative real-time polymerase chain reaction. The vaccine was intramuscularly injected into zebrafish (Danio rerio), and 28 days after immunization, viral hemorrhagic septicemia virus (105 TCID50/10 µl/fish) was intraperitoneally injected. A survival rate of 68% was observed in the pVG + CCL19 group but this was not significantly different from the survival rate of fish treated with pVG alone, that is, without the adjuvant. However, the expression of interferon- and cytokine-related genes in the spleen and kidney tissues of zebrafish was significantly increased (p < 0.05) on days 1, 3, 7, and 14 after immunization. Thus, CCL19a.2 induced an initial immune response as a molecular adjuvant, which may provide initial protection against virus infection before vaccination-induced antibody formation. This study provides insights on the functions of CCL19a.2 adjuvant in DNA vaccines.

Keywords: DNA vaccine; chemokine ligand 19 (CCL19); molecular adjuvant; viral hemorrhagic septicemia virus (VHSV); zebrafish (Danio rerio).

MeSH terms

  • Adjuvants, Immunologic
  • Animals
  • Cytokines
  • Fish Diseases* / prevention & control
  • Glycoproteins / genetics
  • Hemorrhagic Septicemia, Viral* / prevention & control
  • Interferons
  • Ligands
  • Novirhabdovirus* / genetics
  • RNA, Messenger
  • Vaccines, DNA* / genetics
  • Vaccines, DNA* / pharmacology
  • Viral Vaccines* / genetics
  • Zebrafish / genetics

Substances

  • Adjuvants, Immunologic
  • Cytokines
  • Glycoproteins
  • Ligands
  • RNA, Messenger
  • Vaccines, DNA
  • Viral Vaccines
  • Interferons