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Immunological characterization of chitosan adjuvanted outer membrane proteins of Salmonella enterica serovar Typhi as multi-epitope typhoid vaccine candidate

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Abstract

Background

Outer membrane proteins (OMPs) of Gram-negative bacteria have been known as potential vaccine targets due to their antigenic properties and host specificity. Here, we focused on the exploration of the immunogenic potential and protective efficacy of total OMPs of Salmonella enterica serovar Typhi due to their multi epitope properties, adjuvanted with nanoporous chitosan particles (NPCPs).

The study was designed to extrapolate an effective, low cost prophylactic approach for typhoid fever being getting uncontrolled in Pakistan due to emergence of extensively drug resistant (XDR) strains.

Methods & results

The OMPs of two S. Typhi variants (with and without Vi capsule) alone and with nanoporous chitosan particles as adjuvant were comparatively analyzed for immunogenic potential in mice. Adaptive immunity was evaluated by ELISA and relative quantification of cytokine gene expression (IL4, IL6, IL9, IL17, IL10, TNF, INF and PPIA as house keeping gene) using RT-qPCR. Statistical analysis was done using Welch’s test. The protection was recorded by challenging the immunized mice with 50% ×LD50 of S. Typhi. The Vi + ve-OMPs of S. Typhi showed the most promising results by ELISA and significantly high expression of IL-6, IL-10 and IL-17 and 92.5% protective efficacy with no detectable side effects.

Conclusion

We can conclude that the OMPs of Vi + ve S. Typhi are the most promising candidates for future typhoid vaccines because of cost effective preparation without expensive purification steps and multi-epitope properties. Chitosan adjuvant may have applications for oral protein based vaccines but found less effective in injectable preparations.

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Acknowledgements

The acknowledgements are due to National Institute for Biotechnology and Genetic Engineering College (NIBGE-C), PIEAS, Faisalabad, Pakistan for technical facilities to conduct this research project.

Funding

The funding was provided by Higher Education Commission of Pakistan provided funding under the research grant NRPU No. 1771, to support this research work.

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Authors and Affiliations

  1. National Institute for Biotechnology & Genetic Engineering College (NIBGE-C), Faisalabad, Pakistan

    Ambreen Ayub, Muhammad Usman, Ayesha Ihsan, Quratul Ain, Asad Bashir Awan, Muhammad Wajid, Aamir Ali, Mazhar Iqbal & Yasra Sarwar

  2. Institute of Engineering & Applied Sciences (PIEAS), Islamabad, Pakistan

    Ambreen Ayub, Muhammad Usman, Ayesha Ihsan, Quratul Ain, Asad Bashir Awan, Muhammad Wajid, Aamir Ali, Mazhar Iqbal & Yasra Sarwar

  3. Akhuwat Faisalabad Institute For Research In Science & Technology, Faisalabad, Pakistan

    Abdul Haque

  4. Faculty of Veterinary and Animal Sciences, Muhammad Nawaz Shareef University of Agriculture, Multan, Pakistan

    Muhammad Usman

Authors
  1. Ambreen Ayub
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  2. Muhammad Usman
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  3. Ayesha Ihsan
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  4. Quratul Ain
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  7. Aamir Ali
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  8. Abdul Haque
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  9. Mazhar Iqbal
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  10. Yasra Sarwar
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Contributions

Yasra Sarwar and Abdul Haque conceived the idea, developed the research, and finalized the manuscript. Ambreen Ayub (first author) performed the experiments, analyzed the data and wrote the manuscript. Yasra Sarwar and Aamir Ali helped in ELISA and animal studies, including sample collection and processing. Quratul ain, Asad Bashir and Muhammad Wajid helped in qPCR studies and statistical analysis. Muhammad Usman and Ayesha Ihsan prepared and characterized NPCP for adjuvant preparation. Yasra Sarwar, Abdul Haque and Mazhar Iqbal edited and proof read the manuscript. All authors read and agreed the manuscript for publication.

Corresponding author

Correspondence to Yasra Sarwar.

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Conflict of interest

The authors have no relevant financial or non-financial interests to disclose.

Ethics approval

The animal study was reviewed and approved on 26-07-2019 (ID # 193) by The Institutional Animal House Committee at National Institute for Biotechnology and Genetic Engineering (NIBGE), Faisalabad, Pakistan.

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Ayub, A., Usman, M., Ihsan, A. et al. Immunological characterization of chitosan adjuvanted outer membrane proteins of Salmonella enterica serovar Typhi as multi-epitope typhoid vaccine candidate. Mol Biol Rep 49, 7377–7387 (2022). https://doi.org/10.1007/s11033-022-07531-w

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  • DOI: https://doi.org/10.1007/s11033-022-07531-w

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