Construction and comparative study of monovalent and multivalent DNA vaccines against Streptococcus iniae

Streptococcus iniae is an important fish pathogen with a broad host range that includes both marine and freshwater fish species. With an aim to develop effective vaccines against S. iniae, we in this study constructed three monovalent DNA vaccines, i.e., pSagF, pSagG, and pSagI, based on sagF, G, and I, which are components of the streptolysin S cluster. The immunoprotective potentials of these vaccines were examined in a model of Japanese flounder (Paralichthys olivaceus). The results showed that following intramuscular administration, the vaccine plasmids were transported to spleen, kidney, and liver, where the vaccine-encoding transgenes were expressed. Immunocolloidal gold electron microscopy detected production of the vaccine protein in fish vaccinated with each of the vaccine plasmids. Following lethal-dose S. iniae challenge, pSagF-, pSagG-, and pSagI-vaccinated fish exhibited relative percent of survival (RPS) rates of 78%, 65%, and 76% respectively. To examine whether multivalent vaccines composed of different combinations of monovalent vaccines would produce better protections, flounder were vaccinated with FG (pSagF plus pSagG), FI (pSagF plus pSagG), or FGI (pSagF plus pSagG and pSagI). Subsequent challenging study showed that the RPS rates of the fish vaccinated with the divalent and trivalent vaccines were 4%-17% and 13%-26% respectively higher than those of the fish vaccinated with the component monovalent vaccines. Furthermore, FGI exhibited a strong cross protection against both serotype I and serotype II S. iniae, apparently due to, as revealed by sequence analysis, the existence of highly conserved SagF, SagG, and SagI homologs in these serotypes. Immunological analysis showed that all vaccines induced (i) specific serum antibody production, (ii) enhanced complement-mediated bactericidal activity, and (iii) significant induction of a wide range of immune genes. However, the levels of gene expression and serum bactericidal activity induced by FGI were in general more potent than those induced by monovalent vaccines. Taken together, these results indicate that the DNA vaccines based on sagF, G, and I, especially when they are formulated as multivalent vaccines, are highly efficacious against S. iniae infection.