Numbers of predicted CTL, HTL, and BCL epitopes.

<div><p>Significant progress has been made in <i>HIV-1</i> research; however, researchers have not yet achieved the objective of eradicating <i>HIV-1</i> infection. Accordingly, in this study, eucaryotic and procaryotic in silico vaccines were developed for <i>HIV-Gag</i> polyproteins from 100 major <i>HIV</i> subtypes and CRFs using immunoinformatic techniques to simulate immune responses in mice and humans. The epitopes located in the conserved domains of the <i>Gag</i> polyprotein were evaluated for allergenicity, antigenicity, immunogenicity, toxicity, homology, topology, and IFN-γ induction. Adjuvants, linkers, CTLs, HTLs, and BCL epitopes were incorporated into the vaccine models. Strong binding affinities were detected between HLA/MHC alleles, TLR-2, TLR-3, TLR-4, TLR-7, and TLR-9, and vaccine models. Immunological simulation showed that innate and adaptive immune cells elicited active and consistent responses. The human vaccine model was matched with approximately 93.91% of the human population. The strong binding of the vaccine to MHC/HLA and TLR molecules was confirmed through molecular dynamic stimulation. Codon optimization ensured the successful translation of the designed constructs into human cells and <i>E</i>. <i>coli</i> hosts. We believe that the <i>HIV-1 Gag</i> vaccine formulated in our research can reduce the challenges faced in developing an <i>HIV-1</i> vaccine. Nevertheless, experimental verification is necessary to confirm the effectiveness of these vaccines in these models.</p></div>