Fig 1.
Diagrammatic depiction of strategy adopted in the current study.
Table 1.
Final selected T-cell epitopes from HTLV-1 antigenic proteins employed to develop the MEBV construct and their binding details with their corresponding HLA alleles.
Table 2.
Final chosen LBL epitopes from HTLV-1 antigenic proteins employed to develop the MEBV construct.
Fig 2.
Docked complex of chosen 15 epitopes (blue licorice illustration) and their corresponding HLAs (red cartoon illustration) as demonstrated in Table 1.
Fig 3.
Collective population coverage of the chosen T-cell epitopes based on their corresponding HLA alleles.
Areas of specific significance were considered in this display.
Fig 4.
Graphical representation of constructed MEBV structure, encompassing 382 amino acids having one adjuvant (at N-terminal), 9 CTL, 6 HTL and 5 LBL epitopes.
Fig 5.
(A) MEBV construct sequence. Black letters represents the epitopes sequences. Purple letters depict adjuvant sequence, Maroon color shows EAAAK linker, Blue color displays GPGPG linkers and red color demonstrates the KK linkers; (B) Pipes representation of refined 3D model of final MEBV construct (cyan color shows ɑ-helix, red color depicts β-strands and magenta color illustrates random loops); (C) Ramachandran plot analysis of estimated MEBV model, showing the presence of 89.2% residues in the favored region.
Fig 6.
Disulfide engineering, depicting one mutated residues pair with red color.
Fig 7.
Conformational B-cell epitopes (cyan) recognized in the final MEBV vaccine (magenta).
Fig 8.
TLR3-MEBV docked complex shown at the left in cartoon representation.
Interacting residues of MEBV are highlighted at right side. MEBV construct revealed with blue color and TLR3 displayed with red color. Red lines represent salt bridges, blue lines show Hydrogen bonds, whilst orange lines demonstrate the other contacts in docked complex. The colors of interacting residues are interpreting the characteristics of amino acids (neutral: green, Cys: yellow, aromatic: pink, aliphatic: grey, positive: blue, negative: red, and Pro&Gly: orange).
Table 3.
Data of the top TLR3-MEBV docked cluster.
Fig 9.
In silico immune response employing MEBV as an antigen.
(A) Production of immunoglobulin as a result of antigen injection (B) population of B cells after three times antigen exposure.
Fig 10.
In silico cloning of codon optimized MEBV construct into pET28a(+) expression system.
The black region exhibits the plasmid back-bone, whereas the maroon section demonstrates the inserted DNA sequence.
Fig 11.
Comparisons of the RMSD, RMSF values and the number of hydrogen bonds present in the structure of MEBV-TLR3 and TLR3 protein, obtained through MD simulation.
(A) RMSD of the backbone of the MEBV-TLR3 and the TLR3 Protein (B) RMSF of the backbone of the complex and the TLR3 protein (C) Hydrogen bonds present in the MEBV-TLR3 complex and the TLR3 Proteins.