A newly published study in Biochemistry and Biophysics Reports from researchers at the University of Dhaka, Bangladesh, presents a promising immunoinformatics-based approach for developing a multi-epitope vaccine (MEV) targeting the monkeypox virus (MPXV). With global mpox cases exceeding 124,000 as of late 2024, and the emergence of a more virulent Clade Ib in Central Africa, the research addresses an urgent need for a virus-specific vaccine—one that goes beyond repurposed smallpox formulations.
This vaccine design leverages computational tools, including reverse vaccinology and immune simulations, to create a potent, stable, and immunogenic construct. Notably, the study incorporates experimentally validated T-cell epitopes from related orthopoxviruses and conservatively selected B-cell targets from key MPXV glycoproteins involved in viral entry.
Key Findings: A Computational Vaccine Blueprint Against MPXV
A Multi-Pronged Immune Strategy
The vaccine construct incorporates:
- 4 B-cell epitopes from six conserved MPXV membrane glycoproteins, such as H3L and A35R, crucial for viral attachment and host cell entry.
- 9 cytotoxic T lymphocyte (CTL) and 7 helper T lymphocyte (HTL) epitopes, selected from experimentally validated orthopoxvirus sequences with demonstrated immune relevance.
These epitopes were joined with flexible linkers and immunostimulatory adjuvants, including β-defensin 3, PADRE, and Hp91, to enhance immune activation and structural stability.
Strong Interaction with Key Immune Receptors
Molecular docking and simulation revealed that the vaccine binds robustly with Toll-like receptors TLR-2 and TLR-4, key initiators of innate immune responses. HADDOCK scores and low root-mean-square deviation (RMSD) values indicated a stable and favorable interaction profile, a critical step for triggering both arms of the immune system.
Promising Immune Simulation Results
In silico immune simulations projected:
- Elevated IgM during the primary response, followed by a doubling of IgG and IgM+IgG levels in later stages.
- Sustained expansion of helper and cytotoxic T-cell memory populations.
- Elevated interferon-gamma (IFN-γ) and interleukin-2 (IL-2), cytokines critical for long-term immunity and viral control.
Implications for Public Health and National Security
The rapid spread and re-emergence of mpox—as witnessed during the 2022 global outbreak and the more recent 2024–2025 Clade Ib surge—highlight the vulnerabilities in current pandemic response systems. Existing vaccines like JYNNEOS and ACAM2000, developed for smallpox, offer limited protection against MPXV and are not fully approved or optimized for safety and efficacy across populations.
This computational vaccine design represents a scalable, adaptable platform that can respond to viral evolution and offer targeted protection against mpox variants. If validated in experimental systems, this approach could support pre-emptive biodefense strategies, bolster global vaccine equity, and enhance national pandemic readiness by enabling quicker transitions from design to production—especially in resource-limited settings.
Challenges and Next Steps
While the computational data are compelling, the study acknowledges key limitations:
- No in vitro or in vivo validation of immunogenicity or safety has been conducted to date.
- Expression was modeled in E. coli, but real-world protein folding and post-translational modifications may differ.
- Stability and solubility of the full construct in biological systems remain to be confirmed.
Experimental validation in cell lines and animal models is urgently needed to move this vaccine candidate toward clinical development.
This study exemplifies how immunoinformatics and reverse vaccinology can accelerate rational vaccine development against emerging infectious threats. By targeting the monkeypox virus with a multi-epitope design grounded in both experimental epitope data and robust computational modeling, the research sets the stage for new, precision-engineered countermeasures tailored to evolving viral landscapes.
Note: The disease caused by monkeypox virus is now officially called mpox, following a 2022 renaming by the World Health Organization to reduce stigma and improve public communication. While mpox refers to the illness, monkeypox virus (MPXV) remains the accepted scientific name for the virus itself.
Raj KH, Hossain E, Zahin H, et al. A robust comprehensive immunoinformatics approach for designing a potential multi-epitope based vaccine against a reiterated monkeypox virus. Biochemistry and Biophysics Reports. Sep 2025