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Updated: Aug 3, 2025
Expression and Purification of Virus-like Particles for Vaccination
Published on: June 2, 2016
Cloning, expression and in vitro validation of chimeric multi epitope vaccine candidate against visceral
Rupal Ojha1, Kailash Chand2, Balachandar Vellingiri3
1Department of Biochemistry, School of Life Sciences, Central University of Rajasthan, Ajmer 305817, Rajasthan, India.
Abstract:
Visceral Leishmaniasis or Kala-Azar is one of the most severe and deadly neglected tropical disease caused by the Leishmania parasite. A few number of vaccines are going through different phases in clinical trial but failing of these vaccines in successive phase trial or less efficacy, urge to develop highly immunogenic and cost-effective treatment to get rid of deadly VL. This study focuses on the development of more potent vaccine candidate against VL. The recombinant vaccine candidate LeiSp was expressed in Pichia pastoris, followed by purification and characterization. The purified protein was also tested for any post-translation modification, which favors being a potent immunogenic candidate. Further, the expression modulation of different pro-inflammatory and anti-inflammatory cytokines was evaluated in THP1 cell lines. A significant upregulation in the expression of pro-inflammatory cytokines while no significant changes were observed in the expression of anti-inflammatory cytokines. The impact of recombinant vaccine protein candidates in infected conditions were determined. Here, upon treatment with chimeric vaccine protein candidate, we observed a considerable recovery in the expression level of pro-inflammatory cytokines, which were downregulated upon infection alone. In addition to this, we found a significant decrease in the expression of anti-inflammatory cytokines, which were upregulated during infection alone. We further validated our findings in infected hPBMCs and observed similar expression modulation of pro-inflammatory and anti-inflammatory cytokines with and without treatment. Thus, the present study indicates that the chimeric LeiSp protein which was designed using bioinformatics approaches shows a potential inductive efficacy for pro-inflammatory cytokines in Leishmania-infected cells.
Insights
Developing a new recombinant vaccine candidate, LeiSp, shows promise for treating visceral leishmaniasis (Kala-Azar). This potential vaccine effectively modulates immune responses in Leishmania-infected cells, offering hope against this deadly neglected tropical disease.
Area of Science:
- Parasitology and Immunology
- Vaccine Development
- Tropical Diseases
Background:
- Visceral leishmaniasis (Kala-Azar) is a severe, deadly neglected tropical disease caused by Leishmania parasites.
- Existing vaccines face challenges with efficacy and progression through clinical trials, necessitating novel immunogenic and cost-effective treatments.
- There is a critical need for improved therapeutic strategies to combat visceral leishmaniasis.
Purpose of the Study:
- To develop a more potent vaccine candidate against visceral leishmaniasis.
- To express, purify, and characterize a recombinant vaccine candidate, LeiSp.
- To evaluate the immunogenic potential and cytokine modulation effects of the LeiSp vaccine candidate in Leishmania-infected cells.
Main Methods:
- Recombinant LeiSp protein was expressed in Pichia pastoris and purified.
- Post-translational modifications of the purified protein were analyzed.
- Cytokine expression (pro-inflammatory and anti-inflammatory) was evaluated in THP1 cell lines and human peripheral blood mononuclear cells (hPBMCs) from infected individuals, with and without treatment with the vaccine candidate.
Main Results:
- The purified LeiSp protein exhibited characteristics favorable for immunogenicity.
- Treatment with the chimeric LeiSp protein significantly upregulated pro-inflammatory cytokines and downregulated anti-inflammatory cytokines in infected THP1 cells and hPBMCs.
- These modulations represent a recovery from the cytokine dysregulation observed during Leishmania infection alone.
Conclusions:
- The chimeric LeiSp protein, designed using bioinformatics, demonstrates potential inductive efficacy for pro-inflammatory cytokines in Leishmania-infected cells.
- The study indicates that LeiSp is a promising candidate for a visceral leishmaniasis vaccine.
- Further development of this vaccine candidate could offer a new therapeutic avenue for visceral leishmaniasis.
