Immunomodulatory activity of polycaprolactone nanoparticles with calcium phosphate salts against Leishmania infantum infection

Abstract

Objective: To prepare and characterize polycaprolactone (PCL) nanoparticles loaded with sonicator fragmented (SLA) and freeze-thaw Leishmania antigens (FTLA) and to investigate the in vitro immunogenicity of antigen-encapsulated nanoparticles with calcium phosphate adjuvant. Methods: The water/oil/water binary emulsion solvent evaporation method was used to synthesize antigen-loaded PCL nanoparticles. Particles were characterized by scanning electron microscopy and zeta potential measurements. Their cytotoxicity in J774 macrophages in vitro was determined by MTT analysis. In addition, the amount of nitric oxide and the level of cytokines produced by macrophages were determined by Griess reaction and ELISA method, respectively. The protective effect of the developed formulations was evaluated by determining the infection index percentage in macrophages infected with Leishmania infantum. Results: Compared to the control group, SLA PCL and FTLA PCL nanoparticles with calcium phosphate adjuvant induced a 6- and 7-fold increase in nitric oxide, respectively. Additionally, the vaccine formulations promoted the production of IFN-γ and IL-12. SLA PCL and FTLA PCL nanoparticles combined with calcium phosphate adjuvant caused an approximately 13- and 11-fold reduction in infection index, respectively, compared to the control group. Conclusions: The encapsulation of antigens obtained by both sonication and freeze-thawing into PCL nanoparticles and the formulations with calcium phosphate adjuvant show strong in vitro immune stimulating properties. Therefore, PCL-based antigen delivery systems and calcium phosphate adjuvant are recommended as a potential vaccine candidate against leishmaniasis.

Keywords

Leishmaniasis / Calcium phosphate / Polycaprolactone / Nanoparticle / Antigen delivery system / Adjuvant / Vaccine design

Conflict of interest statement

The authors declare that they have no conflict of interest.

Funding

The authors received no extramural funding for the study.

Data availability statement

The data supporting the findings of this study are available from the corresponding authors upon request.

Authors’ contributions

KK and MI conducted the experimental study and analyzed the data. KK prepared the draft and revised the manuscript. EŞA and AA developed the work. EŞA supervised the study. AA and MB reviewed the draft text. All authors read and approved the final version of the manuscript.

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