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 | ||
| مقاله 9، دوره 22، شماره 8، آبان 2019، صفحه 901-907 اصل مقاله (708.92 K) | ||
| نوع مقاله: Original Article | ||
| شناسه دیجیتال (DOI): 10.22038/ijbms.2019.34872.8276 | ||
| نویسندگان | ||
| Fatemeh Ghafarifar* 1؛ Mohammad Jafarimodrek2؛ Hossein Vazini3؛ Zohreh Sharifi4؛ Abdolhossein Dalimi1؛ Mohammad Saaid Dayer1 | ||
| 1Department of Parasitology and Entomology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran | ||
| 2Department of Medical Parasitology, Faculty of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran | ||
| 3Nursing Department, Basic Sciences faculty, Hamedan Branch, Islamic Azad University, Hamedan, Iran | ||
| 4Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran | ||
| چکیده | ||
| 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 µg), pc-MIC3 (50 µg), or combined pCAGGS-IL12 (50 µg) and pc-MIC3 (50 µg). 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 µg) 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. | ||
| کلیدواژهها | ||
| BALB/c mice؛ DNA vaccine؛ Immunization؛ pCAGGS-IL12؛ pc-MIC3؛ Toxoplasma gondii | ||
| مراجع | ||
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