Prioritization of potential vaccine targets using comparative proteomics and designing of the chimeric multi-epitope vaccine against Pseudomonas aeruginosa

摘要

Multidrug-resistant Pseudomonas aeruginosa is one of the worldwide health problems involved inelevated mortality and morbidity. Therefore, it is important to fnd a therapeutic for this pathogen.In the present study, we have designed a chimeric vaccine against P. aeruginosa with the help ofcomparative proteomics and reverse vaccinology approaches. Using comparative subtractive proteomicanalysis of 1,191 proteomes of P. aeruginosa, a total of twenty unique non-redundant proteomeswere selected. In these proteomes, ffteen outer membrane proteins (OMPs) of P. aeruginosa wereselected based on the basis of hydrophilicity, non-secretory nature, low transmembrane helix (1),essentiality, virulence, pathway association, antigenic, and protein-protein network analysis. Reversevaccinology approach was used to identify antigenic and immunogenic MHC class I, MHC class II and Bcell epitopes present in the selected OMPs that can enhance T cell and B cell mediated immunogenicity.The selected epitopes were shortlisted based on their allergenicity, toxicity potentials, solubility, andhydrophilicity analysis. Immunogenic peptides were used to design a multi-epitope vaccine construct.Immune-modulating adjuvants and PADRE (Pan HLA-DR epitopes) sequence were added with epitopessequence to enhance the immunogenicity. All the epitopes, adjuvants and PADRE sequence werejoined by linkers. The designed vaccine constructs (VT1, VT2, VT3, and VT4) were analyzed by theirphysiochemical properties using diferent tools. Selected chimeric vaccine constructs (VT1, VT3, andVT4) were further shortlisted by their docking score with diferent HLA alleles. The fnal selected VT4construct was docked with TLR4/MD2 complex and confrmed by molecular dynamics simulationstudies. The fnal vaccine VT-4 construct was in-silico cloned in pET28a. Therefore, the designedconstruct VT4 may be studied to control the interaction of P. aeruginosa with host and infection causedby P. aeruginosa.