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Immunogenic insights and experimental validation of a multiepitope vaccine construct against Streptococcus pneumoniae
Yogeshwar Devarakonda1, M V N Janardhan Reddy1, Arunima Binu1
1Genetics and Molecular Microbiology Laboratory, Department of Biological Sciences, Birla Institute of Technology & Science, Pilani, Hyderabad, Telangana, India.
Insights
This study experimentally validates a novel multi-epitope vaccine against Streptococcus pneumoniae, demonstrating its safety and ability to elicit a robust immune response, including antibody production and cytokine release.
Area of Science:
- Bacteriology
- Vaccinology
- Immunology
Background:
- *Streptococcus pneumoniae* is a leading cause of severe infections like pneumonia and meningitis, particularly in vulnerable populations.
- *Antibiotic resistance* and the emergence of non-vaccine serotypes diminish the effectiveness of current treatments and vaccines.
- Existing polysaccharide vaccines offer limited serotype coverage, necessitating the development of broader-protection strategies.
Purpose of the Study:
- To experimentally validate a computationally designed multi-epitope vaccine candidate against *Streptococcus pneumoniae*.
- To assess the safety, immunogenicity, and efficacy of this novel vaccine construct.
Main Methods:
- The multi-epitope vaccine construct was expressed in *Escherichia coli* and purified using Ni-NTA affinity chromatography.
- Protein characterization involved SDS-PAGE, Western blot, and circular dichroism spectroscopy.
- Immunogenicity and safety were evaluated in BALB/c mice through antibody response assays (Western blot, ELISA), cytokine profiling, and hemolysis assays.
Main Results:
- The purified vaccine protein's structure aligned with computational predictions.
- Hemolysis assays confirmed the vaccine's safety and tolerability.
- Administration in mice induced significant antibody titers and a broad cytokine response, including interleukins, TNF-α, and GM-CSF.
Conclusions:
- This study presents the first experimental validation of a designed multi-epitope vaccine against *S. pneumoniae*.
- The vaccine demonstrated promising immunogenicity, characterized by antibody production and a significant cytokine profile.
- The absence of hemolytic toxicity supports its potential as a safe and effective vaccine candidate.
Abstract:
Streptococcus pneumoniae is a predominant cause of pneumonia, sinusitis, bacteremia, meningitis and otitis media especially in children and the elderly. The emergence of antibiotic resistance and shift towards non-vaccine serotypes has compromised the efficacy of the treatment regime and available vaccines. The available polysaccharide-based vaccines confer protection against limited serotypes. In this study, we have conducted the experimental validation of a promising multi epitope vaccine candidate. The vaccine construct was cloned into pET-28a vector and expressed in Escherichia coli. The expression of the protein was induced, and its purification was carried out by the Ni-NTA affinity chromatography. Further, purified multiepitope protein was subjected to the SDS-PAGE and western blot analysis. The secondary structure was determined by circular dichroism spectroscopy, and it was observed to closely align with the predicted structure by Alpha fold. The hemolysis assay confirmed its safety and tolerability. The BALB/c mice was administered with this multiepitope vaccine construct and immune response was evaluated. The antibody response was estimated and confirmed by western blot and ELISA analysis. The cytokine profiling assay revealed the involvement of interleukins, TNF-α, and GM-CSF. Together, this study provides the first experimental validation of a computationally designed multi-epitope vaccine against S. pneumoniae, demonstrating antibody titers, broad cytokine response, and absence of hemolytic toxicity.
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