Fig 1.
Systematic workflow of the study design.
Each colour represents the specific step which can be recognized by the colour scheme given in the right panel.
Table 1.
Summary of selected CTL epitopes for envelope and nucleoprotein of Hantavirus.
Table 2.
Summary of Helper T-cell epitopes selected for envelope and nucleoprotein of Hantavirus using the IEDB MHC-II module.
Table 3.
Results of ABCPred: Summary of linear B cell epitopes.
Fig 2.
Topological representation of the vaccine constructs.
(a) represent the topological architecture of the Env-Vac construct from envelope polyprotein, (b) represent the topological arrangement of the predicted epitopes from nucleoprotein NP-Vac construct while (c) represents the topological organization of the epitopes from different proteins named as Com-Vac.
Fig 3.
Structural modeling and epitopes mapping for each construct.
(A) represent the 3D structure of envelope protein-based MEVC where different colour represent different epitopes. (B) represent the 3D structure of envelope protein-based MEVC where different colour represent different epitopes. (C) represent the 3D structure of envelope protein-based MEVC where different colour represent different epitopes.
Fig 4.
Structural validation of the modeled vaccine constructs.
(A-C) show the Ramachandran plots for all the designed vaccine constructs. (D-E) show the ProSA-web results for all the designed vaccine constructs.
Fig 5.
Secondary structural elements distribution in each vaccine construct.
(A) show the secondary structure for Env-Vac, (B) show the SS for NP-Vac while (C) show the SS for Com-Vac.
Fig 6.
Solubility analysis of each vaccine construct.
(A) show the solubility graph for Env-Vac, (B) show the solubility graph for NP-Vac and (C) show the solubility graph for Com-Vac.
Table 4.
The physiochemical properties of constructed vaccines.
Fig 7.
The predicted conformational B cell epitopes for each vaccine construct.
(a-d) shows the CBCEs in Env-Vac, (e-i) shows the CBCEs in NP-Vac while (j-n) show the CBCEs in Com-Vac.
Table 5.
The predicted B cell conformational epitopes with their residues, size and scrores.
Fig 8.
Interaction pattern of the designed vaccine constructs and TLR3.
(a-b) show the binding mode of Env-Vac with TLR3, (c-d) show the interaction pattern of NP-Vac-TLR3 and (e-f) show the interaction pattern of com-Vac with TLR3.
Table 6.
Summary of binding free energies for docked complexes (Multi-Epitopes Vaccine constructs-TLR3).
Fig 9.
Cloning the optimized vaccine sequences into the pET28a (+) expression system.
(a) envelope vaccine sequence was cloned into the pET28a (+) vector, (b) nucleoprotein vaccine sequence was cloned into the pET28a (+) vector, (c) protein-wide vaccine sequence was cloned into the pET28a (+) vector.
Fig 10.
Immune simulation of the designed vaccine against the Hantavirus.
(a-c) show the predicted antibodies response after three doses while (d-f) show the secondary immune elements response upon the injection.