Retrospective Computational and Immunoinformatics Validation of Clinically Evaluated Necator americanus Protein Vaccine Candidates: Na-APR-1, Na-GST-1, and Na-ASP-2

Hookworm disease, caused by Necator americanus, impacts approximately 472 million individuals globally, resulting in significant public health burden. Currently, there is no approved vaccine for this condition, leading researchers to explore computational and immunoinformatics validation of existing vaccine candidates. Three such candidates, Na-APR-1, Na-GST-1, and Na-ASP-2, underwent retrospective evaluation for their safety and immunogenicity.

Predictive modeling identified Na-ASP-2 as both toxigenic and allergenic, mirroring its previous clinical failure. In contrast, Na-APR-1 demonstrated a favorable safety profile, while Na-GST-1 displayed inconsistent allergenicity signals, necessitating further experimental validation. All candidates exhibited a rich repertoire of epitopes capable of stimulating CD8+ T cells (CTL), helper T cells (HTL), B cells, and cytokine production. Na-APR-1, in particular, displayed the broadest range of such immunogenic elements.

Broad global HLA population coverage across endemic regions was also observed, suggesting potential applicability of all candidates. Molecular docking simulations revealed that all antigens interacted favorably with TLR4, the Toll-like receptor, with Na-GST-1 and Na-APR-1 exhibiting the strongest predicted binding. Normal mode analyses further indicated stable antigen-TLR4 complex dynamics across all candidates.

Immune simulations predicted robust, long-lasting humoral and cellular immune responses from Na-APR-1 and Na-ASP-2, while Na-GST-1 was predicted to have significantly reduced immunogenicity. These findings validate a computational framework for advancing rational hookworm vaccine design, with Na-APR-1 as a strong candidate for continued clinical development, and Na-GST-1 requiring further immunogenic verification.