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

• Published in: Experimental parasitology Vol. 118; no. 3; pp. 393 - 401
• Main Authors: Rafati, Sima, Zahedifard, Farnaz, Azari, Mahnaz Kakeh, Taslimi, Yasaman, Taheri, Tahereh
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 01.03.2008; Elsevier
• Subjects: Animals; Antibodies, Protozoan - biosynthesis; Antibodies, Protozoan - blood; Biological and medical sciences; Blotting, Western; bovine serum albumin; BSA; C-terminal extension of cysteine proteinase type I; CMV; Con A; concanavalin A; CpG; Cysteine Endopeptidases - chemistry; Cysteine Endopeptidases - genetics; Cysteine Endopeptidases - immunology; cytomegalovirus; cytosine phosphate guanine oligodeoxynucleotide; deoxyribonucleic acid; DHFR; dihydrofolate reductase; DNA; E. coli; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Escherichia coli; F/T; Female; frozen-and-thawed; Fundamental and applied biological sciences. Psychology; His; histidine; IFN-γ; IgG; IL-5; Immunization, Secondary; Immunogenicity; immunoglobulin G; Immunoglobulin G - biosynthesis; Immunoglobulin G - blood; Immunology; interferon gamma; Interferon-gamma - analysis; interleukin 5; Interleukin-5 - analysis; IPTG; isopropyl β-d-1-thiogalactopyranoside; L. infantum; Leishmania infantum; Leishmania infantum - enzymology; Leishmania infantum - genetics; Leishmania infantum - immunology; Leishmaniasis, Visceral - immunology; Leishmaniasis, Visceral - prevention & control; Life cycle. Host-agent relationship. Pathogenesis; Life Sciences; Liver - parasitology; Luria Bertani; Mice; Mice, Inbred BALB C; nickel–nitrilotriacetic acid; Ni–NTA; O-phenylenediamine; OPD; PBMC; PBS; peripheral blood mononuclear cell; phosphate buffer saline; Protozoa; Protozoan Vaccines - immunology; SDS–PAGE; sodium dodecyl sulfate–polyacrylamide gel electrophporesis; Spleen - immunology; Spleen - parasitology; T helper; Vaccines, DNA - immunology; Vaccinology; PBS; OPD; Con A; IgG; IL-5; F/T; SDS–PAGE; PBMC; CMV; C-terminal extension of cysteine proteinase type I; DHFR; BSA; IFN-γ; His; Ni–NTA; TH; Immunogenicity; DNA; LB; IPTG; CpG; L. infantum; E. coli; Kinetoplastida; Protozoa; Cysteine; Leishmania infantum; Vaccination; Rodentia; Parasite; Extension; Vertebrata; Mammalia; Mouse
• ISSN: 0014-4894; 1090-2449
• DOI: 10.1016/j.exppara.2007.10.004

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 100μg of pcDNA-CTE and 3 weeks later boosted with combination of 30μg rCTE, 50μg 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.