Assessment of DNA vaccine encoding Toxoplasma gondii microneme complete gene and IL-12 as adjuvant in BALB/c mice

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GHAFFARIFAR FATEMEH; JAFARI MODREK MOHAMMAD; Vazini Hossein; SHARIFI ZOHREH; DALIMI ABDOLHOSSEIN; DAYER MOHAMMAD SAAID

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Journal: Iranian Journal of Basic Medical Sciences Year:2019 | Volume:22 | Issue:8 Page(s): 901-907

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Title

Assessment of DNA vaccine encoding Toxoplasma gondii microneme complete gene and IL-12 as adjuvant in BALB/c mice

Author(s)

Keywords

BALB/c mice DNA vaccine Immunization pCAGGS

IL12 pc

MIC3 Toxoplasma gondii

Abstract

Objective(s): Toxoplasma gondii is an obligate intracellular protozoan parasite that causes toxoplasmosis in humans and animals. Micronemes (MICs) are effective candidates for DNA vaccine. Materials and Methods: In this study, we evaluated the immune response of BALB/c mice against MIC3 gene of Toxoplasma gondii and interleukin 12 (IL-12) as DNA vaccine. The MIC3 gene was cloned into the PTZ57R/T vector before sub-cloning in pcDNA3. Recombinant pc-MIC3 was transformed into Escherichia coli (TOP10 strain). The pc-MIC3 plasmid was then transfected into Chinese Hamster Ovary (CHO) cells, and the expression of the MIC3 gene was evaluated by SDS-PAGE and Western blotting. Sixty female BALB/c mice were divided into 6 groups. Each group received 3 intramuscular immunizations on days 0, 21st and 42nd using one of the following stimulants: phosphate-buffered saline, pcDNA3, pCAGGS-IL12, pc-MIC3 (100 μ g), pc-MIC3 (50 μ g), or combined pCAGGS-IL12 (50 μ g) and pc-MIC3 (50 μ g). The enzyme-linked immunosorbent assays was applied to evaluate interferon gamma (IFN-γ ) and IL-4 cytokines excretion of lymphocytes stimulated with tachyzoites lysate antigen, as well as the total levels of immunoglobulin G (IgG), IgG2a and IgG1 in immunized mice sera. Results: Our results showed that mice challenged with pc-MIC3 (100 μ g) had the highest longevity and quantity of immunoglobulin. Moreover, the highest expression level of IFN-γ was found in mice injected with combined pcMIC3 and pCAGGS-IL12 (P<0. 05). Conclusion: The MIC3 gene can be an efficient DNA vaccine candidate against toxoplasmosis. While, the single-gene vaccine can confer partial protection to mice against toxoplasmosis, the multigene vaccine can significantly enhance immune responses.

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APA: Copy

GHAFFARIFAR, FATEMEH, JAFARI MODREK, MOHAMMAD, Vazini, Hossein, SHARIFI, ZOHREH, DALIMI, ABDOLHOSSEIN, & DAYER, MOHAMMAD SAAID. (2019). Assessment of DNA vaccine encoding Toxoplasma gondii microneme complete gene and IL-12 as adjuvant in BALB/c mice. IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES, 22(8), 901-907. SID. https://sid.ir/paper/725672/en

Vancouver: Copy

GHAFFARIFAR FATEMEH, JAFARI MODREK MOHAMMAD, Vazini Hossein, SHARIFI ZOHREH, DALIMI ABDOLHOSSEIN, DAYER MOHAMMAD SAAID. Assessment of DNA vaccine encoding Toxoplasma gondii microneme complete gene and IL-12 as adjuvant in BALB/c mice. IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES[Internet]. 2019;22(8):901-907. Available from: https://sid.ir/paper/725672/en

IEEE: Copy

FATEMEH GHAFFARIFAR, MOHAMMAD JAFARI MODREK, Hossein Vazini, ZOHREH SHARIFI, ABDOLHOSSEIN DALIMI, and MOHAMMAD SAAID DAYER, “Assessment of DNA vaccine encoding Toxoplasma gondii microneme complete gene and IL-12 as adjuvant in BALB/c mice,” IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES, vol. 22, no. 8, pp. 901–907, 2019, [Online]. Available: https://sid.ir/paper/725672/en

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