Lactococcus lactis expressing sand fly PpSP15 salivary protein confers long-term protection against Leishmania major in BALB/c mice

PLoS Negl Trop Dis. 2020 Jan 3;14(1):e0007939. doi: 10.1371/journal.pntd.0007939. eCollection 2020 Jan.

Abstract

Cutaneous leishmaniasisis a vector-borne disease transmitted by Leishmania infected sand flies. PpSP15 is an immunogenic salivary protein from the sand fly Phlebotomus papatasi. Immunization with PpSP15 was shown to protect against Leishmania major infection. Lactococcus lactis is a safe non-pathogenic delivery system that can be used to express antigens in situ. Here, the codon-optimized Ppsp15-egfp gene was cloned in pNZ8121 vector downstream of the PrtP signal peptide that is responsible for expression and secretion of the protein on the cell wall. Expression of PpSP15-EGFP recombinant protein was monitored by immunofluorescence, flow cytometry and Western blot. Also, expression of protein in cell wall compartment was verified using whole cell ELISA, Western blot and TEM microscopy. BALB/c mice were immunized three times with recombinant L. lactis-PpSP15-EGFPcwa, and the immune responses were followed up, at short-term (ST, 2 weeks) and long-term (LT, 6 months) periods. BALB/c mice were challenged with L. major plus P. papatasi Salivary Gland Homogenate. Evaluation of footpad thickness and parasite burden showed a delay in the development of the disease and significantly decreased parasite numbers in PpSP15 vaccinated animals as compared to control group. In addition, immunized mice showed Th1 type immune responses. Importantly, immunization with L. lactis-PpSP15-EGFPcwa stimulated the long-term memory in mice which lasted for at least 6 months.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Female
  • Insect Proteins / immunology
  • Lactococcus lactis / genetics
  • Lactococcus lactis / metabolism*
  • Leishmania major*
  • Leishmaniasis, Cutaneous / transmission
  • Mice, Inbred BALB C
  • Phlebotomus / genetics
  • Recombinant Proteins / immunology
  • Salivary Proteins and Peptides / genetics
  • Salivary Proteins and Peptides / metabolism*

Substances

  • Insect Proteins
  • Recombinant Proteins
  • Salivary Proteins and Peptides

Grants and funding

The study is funded by Pasteur Institute of Iran (grant number 916 received by TT, http://www.pasteur.ac.ir/) and University of Tabriz (grant number 39/1716 received by ED, http://tabrizu.ac.ir/en). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.