In silico Designed Ebola Virus T-Cell Multi-Epitope DNA Vaccine Constructions Are Immunogenic in Mice

Abstract

Background: The lack of effective vaccines against Ebola virus initiates a search for new approaches to overcoming this problem. The aim of the study was to design artificial polyepitope T-cell immunogens––candidate DNA vaccines against Ebola virus and to evaluate their capacity to induce a specific immune response in a laboratory animal model. Method: Design of two artificial polyepitope T-cell immunogens, one of which (EV.CTL) includes cytotoxic and the other (EV.Th)––T-helper epitopes of Ebola virus proteins was carried out using original TEpredict/PolyCTLDesigner software. Synthesized genes were cloned in pcDNA3.1 plasmid vector. Target gene expression was estimated by synthesis of specific mRNAs and proteins in cells transfected with recombinant plasmids. Immunogenicity of obtained DNA vaccine constructs was evaluated according to their capacity to induce T-cell response in BALB/c mice using IFNγ ELISpot and ICS. Results: We show that recombinant plasmids pEV.CTL and pEV.Th encoding artificial antigens provide synthesis of corresponding mRNAs and proteins in transfected cells, as well as induce specific responses both to CD4+ and CD8+ T-lymphocytes in immunized animals. Conclusions: The obtained recombinant plasmids can be regarded as promising DNA vaccine candidates in future studies of their capacity to induce cytotoxic and protective responses against Ebola virus.

An immunoinformatic approach driven by experimental proteomics: in silico design of a subunit candidate vaccine targeting secretory proteins of Leishmania donovani amastigotes

“…Next phases of the reverse vaccinology approach would ideally involve assessing the recombinant immunogenic protein expressed in the E. coli (strain K12) system as proposed here, in vitro stimulation of peripheral blood mononuclear cells from active VL patients as well as healthy endemic people for cytokine production, and evaluation in challenge models. While a multi-epitope vaccine molecule generated by using a reverse vaccinology approach can induce specific responses in in vivo and in vitro assays, a single recombinant molecule can also reduce the cost of production [93,94]. The in silico designed vaccine reported here confers substantial immunogenic potential to be considered for in vitro experimental evaluation in the next phase of the study.…”

A Candidate Multi-Epitope Vaccine Against Pathogenic Chandipura Vesiculovirus Identified using Immunoinformatics.

“…A similar approach was used by Bazhan and his co-workers, where they have designed a T-cell multi epitope vaccine against Ebola virus. The Tcell epitopes were predicted using IEDB -Immune Epitope Database and the vaccine candidate constructed using the suitable epitopes were found to be immunogenic when expressed in mice [92].…”

An immunoinformatic approach driven by experimental proteomics: in silico design of a subunit candidate vaccine targeting secretory proteins of Leishmania donovani amastigotes

“…For primary activation and maintenance of CD8+ T cells, CD4+ T cells (both Th1 and Th2 type) [86] are believed to be required [87,88], where cytokines such as IFN-γ, IL-2 and IL-4 could be involved [89][90][91][92] and evaluation in challenge models. While a multi-epitope vaccine molecule generated by using a reverse vaccinology approach can induce specific responses in in vivo and in vitro assays, a single recombinant molecule can also reduce the cost of production [93,94]. The in silico designed vaccine reported here confers substantial immunogenic potential to be considered for in vitro experimental evaluation in the next phase of the study.…”