Fig 1.
A summary of the study on the development of a chimeric multiepitope mRNA vaccine.
Table 1.
An inventory of chosen CTL (MHC-I) epitopes derived from the G, F, and M proteins, accompanied by their allergenic, antigenic, and cytotoxic properties.
Table 2.
Evaluation of particular HTL (MHC-II) epitopes derived from the proteins G, F, and M, along with their allergenicity, antigenicity, capacity to induce IFN- and IL-10, and toxicity.
Table 3.
A listing of chosen linear BCL epitopes accompanied by their allergenic, antigenic, and toxicity.
Fig 2.
Vaccine population coverage of the epitopes (CTL, HTL and combined) that qualified for inclusion in the HeV vaccine construct.
Fig 3.
The assembly of a multiepitope vaccine targeting HeV and the ribbon 3° model of the vaccine.
The schematic illustration of a peptide sequence consisting of 494 amino acids include adjuvant (sky blue color), epitopes (CTL-coral orange, HTL-tea green, B-cell-light violet), and linkers (EAAAK-green, AYY-blue, AK-maroon, KFER-orange).
Table 4.
Physicochemical characteristics of the HeV-vaccine.
Fig 4.
(A) The 2° vaccine structure that was identified via PSIPRED where confidence level in the prediction is denoted by the initial bar (Conf), and the magnitude of each subsequent bar corresponds to a distinct level of confidence. In the second bar (Cart), the beta-sheet is denoted by the yellow color, the helix by the pink color, and the vaccine’s coil structure by the grey color. The third bar (Pred) and fourth bar (AA) correspond to three discrete structural elements and sequences of amino acids, respectively, (B) Graphical representation of the 2° structure of vaccine by SOPMA, (C) Presentation of the 2° structure of the vaccine in graphical form by GOR4.
Table 5.
The GOR4 and SOPMA algorithm findings.
Fig 5.
The constructed vaccine was validated using I-TASSER in a model.
(A) Ramachandran plot refinement, (B) ERRAT score, and (C) Z-score.
Fig 6.
3° model showing the conformational B-cell epitopes of the vaccine design.
[A–H] categories. Grey sticks represent the full vaccination process, while yellow surfaces exhibit conformational B cell epitopes. Anatomical B-cell epitope modeling in three dimensions for the vaccine (A-H). Sticks in gray represent the vaccine component, while surfaces in yellow represent conformational B cell epitopes.
Fig 7.
The association of HeV-vaccine (cyan) and receptors (green).
In the first case, we have the vaccine-TLR-2 interaction (A) and in the second case the vaccine-TLR-4 interaction (B) is illustrated.
Table 6.
Docking values and associations between receptors and vaccine complex.
Fig 8.
The structures from “V-apo”, “V-TLR-2”, and “V-TLR-4” were shown in the plots along with their computed RMSD (A, C, and E) and RMSF (B, D, and F) values.
Fig 9.
The Rg (A, D, and G), SASA (B, E, and H) and Hydrogen bonds (C, F and I) values of the “V-apo”, “V-TLR-2”, and “V-TLR-4” are graphically represented.
Fig 10.
Predicted mRNA structure of the vaccine RNAfold web server.
The centroid structure of the vaccine with the base pair probabilities (A), optimal structure with base pair probabilities (C), centroid structure of the vaccine with the positional entropy (B) and optimal structure of the vaccine with positional entropy (D).
Fig 11.
Cloning the pET-28a(+) plasmid vector in a virtual environment.
Plasmid pET-28a(+) was used to insert the vaccination sequence, and the software SnapGene, which is available for free trial at https://www.snapgene.com/free-trial/, was important in this process. The Hendra Vaccine was transformed into DNA (C) using the pET-28a(+) vector (A). The red part of the cloned Hendra vaccine symbolizes the vaccine’s coding genes, while the black part reflects the vector’s backbone.
Fig 12.
A C-ImmSim-predicted vaccination immunological simulation.
In the aftermath of three successive doses, the immune response manifests as different populations of B-cells (A), TH-cells (B), TR-cells (C), NK cells (D), Dendritic cells (E), and Macrophages (F).