Execution and Design of an Anti HPIV-1 Vaccine with Multiple Epitopes Triggering Innate and Adaptive Immune Responses: An Immunoinformatic Approach

Abstract

Human Parainfluenza Virus (HPIV) Type-1, which is an anti-sense ribonucleic acid (RNA) virus belonging to the paramyxoviridae family, induces upper and lower respiratory tract infections. The infections caused by the HPIV Type-1 virus are usually confined to northwestern regions of America. HPIV-1 causes infections through the virulence of the hemagglutinin-neuraminidase (HN) protein, which plays a key role in the attachment of the viral particle with the host’s receptor cells. To the best of our knowledge, there is no effective antiviral drugs or vaccines being developed to combat the infection caused by HPIV-1. In the current study, a multiple epitope-based vaccine was designed against HPIV-1 by taking the viral HN protein as a probable vaccine candidate. The multiple epitopes were selected in accordance with their allergenicity, antigenicity and toxicity scoring. The determined epitopes of the HN protein were connected simultaneously using specific conjugates along with an adjuvant to construct the subunit vaccine, with an antigenicity score of 0.6406. The constructed vaccine model was docked with various Toll-like Receptors (TLRs) and was computationally cloned in a pET28a (+) vector to analyze the expression of vaccine sequence in the biological system. Immune stimulations carried out by the C-ImmSim Server showed an excellent result of the body’s defense system against the constructed vaccine model. The AllerTop tool predicted that the construct was non-allergen with and without the adjuvant sequence, and the VaxiJen 2.0 with 0.4 threshold predicted that the construct was antigenic, while the Toxinpred predicted that the construct was non-toxic. Protparam results showed that the selected protein was stable with 36.48 instability index (II) scores. The Grand average of Hydropathicity or GRAVY score indicated that the constructed protein was hydrophilic in nature. Aliphatic index values (93.53) confirmed that the construct was thermostable. This integrated computational approach shows that the constructed vaccine model has a potential to combat laryngotracheobronchitis infections caused by HPIV-I.

Development of conserved multi-epitopes based hybrid vaccine against SARS-CoV-2 variants: an immunoinformatic approach

“…Nevertheless, to further enhance the potency and longevity of the immune response against the viral particles, an adjuvant, Human Beta Defensin 3, was incorporated at the beginning of the construct to magnify the effects of selected epitopes. As we know the adjuvants are known to modulate the immunogenicity of epitopes while reducing the need for a large number of antigens to induce the same response (Naveed et al, 2022). The antigenicity results were con rmed by immune simulations, which demonstrated the generation of immunogenic cells and molecules that are required for the establishment of long-term immunity against various SARS-COV-2 variants.…”

“…In order to evaluate the antigenicity of the four selected proteins, they were screened on the VaxiJen v2.0 server as this server predicts the antigenicity of proteins by analyzing their physicochemical properties. The recommended threshold value of 0.4 was utilized to analyze the viral proteins, indicating that proteins with scores above this threshold are considered highly antigenic (Doytchinova and Flower, 2007;Naveed et al, 2022).…”

“…The design consisted of a 6X histidine marker, an adjuvant, B-cell-speci c epitopes, and T-cell-speci c epitopes that were connected in a series using speci c linkers. To connect the selected epitopes, we used various linkers, including EAAK for B-cell epitopes, GPGPG for MHC-I epitopes, and AAY for MHC-II epitopes (Naveed et al, 2022). The vaccine construct was designed, to begin with the inclusion of the adjuvant Human Beta Defensin 3 with UniProt ID: Q5U7J2, which has been shown to enhance the immune response to vaccines (Ferris et al, 2013).…”

“…The engagement of Toll-like receptors (TLRs) in the initiation of intracellular signaling pathways that culminate in the release of various immune compounds such as IRF3, NF-κB, in ammatory cytokines, and IL8 during the initial stages of viral infection has been well-established. To further explore this phenomenon, this study used the employed docking analysis of ve TLRs, namely TLR2, TLR3, TLR4, TLR5, and TLR8, with 6NIG, 2A0Z, 4G8A, 3J0A, and 4R0A PDB IDs, respectively (Naveed et al, 2022). The Cluspro 2.0 server was utilized to carry out the docking (Kozakov et al, 2017) and the resulting docked complexes were then analyzed and visualized using the PyMol software.…”

“…For this investigation, simulation steps were adjusted to 1050 in order to achieve the maximum elapsed time. The second modi ed parameter was the Time Steps of Injection for the second and third booster doses, which were adjusted to 84 and 170, respectively (Naveed et al, 2022).…”