弓形虫鸡尾酒DNA疫苗的免疫保护作用研究

doi:10.19640/j.cnki.jtau.2023.03.008

Journal of Tianjin Agricultural University ›› 2023, Vol. 30 ›› Issue (3): 40-46.doi: 10.19640/j.cnki.jtau.2023.03.008

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Study on immune protection of Toxoplasma gondii cocktail DNA vaccine

He Jinling¹, Du Rongqi¹, Zheng Qiyue¹, Song Qiqi¹, Zhang Dongchao^{1,Corresponding Author}, Jin Tianming^{2,Corresponding Author}

1. Tianjin Key Laboratory of Agricultural Animal Breeding and Healthy Husbandry, College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, Tianjin 300392, China;
2. Tianjin Academy of Agricultural Sciences, Tianjin 300192, China

Received:2022-04-26 Online:2023-06-30 Published:2023-09-06

Abstract

Abstract: The aim of this study is to construct a DNA cocktail vaccine against Toxoplasma gondii（T. gondii）and evaluate its immunoprotective effect on mice. The genes encoding T. gondii ROP5, ROP9, and ROP17 were constructed into the eukaryotic expression vector pVAX1, and the recombinant eukaryotic expression plasmids were named pVAX1-ROP5, PVAX1-ROP9, and PVAX1-ROP17, respectively. The target genes of the recombinant plasmids were verified by PCR and double enzyme digestion, and the expression of the plasmids in eukaryotic cells was detected by Western blot. The three recombinant plasmids were mixed at the ratio of 1∶1∶1, subsequently, the 100 μg plasmid mixture was used to immunize mice by intramuscular injection of leg. After the booster immunization, serum antibody, lymphocyte proliferation, and cytokine levels of mice were detected, and T. gondii RH virulent strain was inoculated to evaluate the protection effect of the immunized mice. The results showed that when the eukaryotic expression plasmids pVAX1-ROP5, pVAX1-ROP9, and pVAX1-ROP17 were verified by PCR and double-digestion, the specific target bands were found at 1 686 bp, 1 128 bp, and 1 836 bp, respectively. Western blot analysis showed that the target bands were detected at 62, 41, and 68 kDa, which were consistent with the molecular weight of the expected target proteins ROP5, ROP9, and ROP17, respectively. Compared with pVAX1 and normal saline group, all the levels of antibody IgG in serum, cytokines IFN-γ and IL-6 produced by splenic lymphocytes, and stimulation index（SI）of splenic lymphocyte proliferation produced by mice immunized with cocktail vaccine were extremely significantly increased（P<0.01）. After inoculation with T. gondii RH virulent strain, the survival time of the immunized mice（16.1±2.07）was extremely significantly prolonged（P<0.01）than that of control mice（9.2±0.74 or 9.0±0.77）. The results indicated that the constructed T. gondii cocktail DNA vaccine pVAX1-ROP5/ROP9/ROP17 could stimulate the humoral and cellular immune responses in mice and enhance the resistance of mice to the parasite infection. The results provide reference for the development of the novel T. gondii vaccine.

Key words: Toxoplasma gondii, DNA vaccine, ROP5, ROP9, ROP17

CLC Number:

S855.9

He Jinling, Du Rongqi, Zheng Qiyue, Song Qiqi, Zhang Dongchao, Jin Tianming. Study on immune protection of Toxoplasma gondii cocktail DNA vaccine[J]. Journal of Tianjin Agricultural University, 2023, 30(3): 40-46.

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Study on immune protection of Toxoplasma gondii cocktail DNA vaccine

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