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Leishmania infantum: prime boost vaccination with C-terminal extension of cysteine proteinase type I displays both type 1 and 2 immune signatures in BALB/c mice
Authors:Rafati Sima  Zahedifard Farnaz  Azari Mahnaz Kakeh  Taslimi Yasaman  Taheri Tahereh
Institution:Molecular Immunology and Vaccine Research Laboratory, Pasteur Institute of Iran, Tehran, Iran. sima-rafatisy@pasteur.ac.ir
Abstract:The aims of current study are to describe the immunogenicity and protective efficacy of prime boost vaccine using C-terminal extension (CTE) of cysteine proteinase type I of Leishmania infantum in BALB/c mice. Group I as vaccinated group primed with 100 microg of pcDNA-CTE and 3 weeks later boosted with combination of 30 microg rCTE, 50 microg of CpG and Montanide 720. Groups II and III were served as control groups. Although, this vaccination regimen did not protect mice against the infectious challenge but it was highly immunogenic. IgG2a has been raised strongly against rCTE in contrast to IgG1 and remains high at every time point under study. By analysis of CTE synthetic peptides (CTE100) before challenge, both IgG1 and IgG2a were produced and for all overlapping synthetic peptides (CTE 1-8) IgG1 raised significantly. This statue is changed at 7 weeks after challenge and only CTE2 and CTE3 have shown to induce considerable amount of IgG1. In all groups, the level of IL-5 started to increase with high concentration shortly passing only 3 weeks after infectious challenge. In compare with two control groups, the vaccinated group produced significantly higher level of IL-5 at 7 weeks post-infection. The parasite burden of all groups is similar at 4 weeks post-challenge in both liver and spleen. In contrast, at 8 weeks post challenge, the spleen of the vaccinated group showed significantly higher level of parasite load in compare with two control groups. This study demonstrated that immunization with CTE display both type 1 and 2 immune signatures in experimental murine model of L. infantum infection.
Keywords:IL-5  interleukin 5  IgG  immunoglobulin G  PBMC  peripheral blood mononuclear cell  F/T  frozen-and-thawed  PBS  phosphate buffer saline  CMV  cytomegalovirus  DNA  deoxyribonucleic acid  IFN-γ  interferon gamma  E  coli  Escherichia coli  LB  Luria Bertani  DHFR  dihydrofolate reductase  Ni-NTA  nickel-nitrilotriacetic acid  CpG  cytosine phosphate guanine oligodeoxynucleotide  OPD  O-phenylenediamine  BSA  bovine serum albumin  Con A  concanavalin A  IPTG  d-1-thiogalactopyranoside" target="_blank">isopropyl β-d-1-thiogalactopyranoside  SDS-PAGE  sodium dodecyl sulfate-polyacrylamide gel electrophporesis  His  histidine  TH  T helper
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