DNA prime and peptide boost immunization protocol encoding the Toxoplasma gondii GRA4 induces strong protective immunity in BALB/c mice.
DNA prime and peptide boost immunization protocol encoding the Toxoplasma gondii GRA4 induces strong protective immunity in BALB/c mice.
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编码弓形虫 GRA4 的 DNA 初免和肽加强免疫方案在 BALB/c 小鼠中诱导强保护性免疫
DOI：
10.1186/1471-2334-13-494
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发表时间：
2013-10-23
影响因子：
3.7
通讯作者：
He S
中科院分区：
文献类型：
--
作者：
Meng M;Zhou A;Lu G;Wang L;Zhao G;Han Y;Zhou H;Cong H;Zhao Q;Zhu XQ;He S
弓形虫是一种广泛分布于细胞内的寄生虫，可感染大多数脊椎动物宿主并引起人畜共患感染。疫苗策略仍然是预防和控制弓形虫病的一种有前途的方法。T.弓形虫GRA 4蛋白已被鉴定为疫苗开发的潜在候选物。在我们的研究中，我们评估了编码TgGRA 4的四种不同免疫接种策略诱导的免疫应答。BALB/c小鼠肌内（i. m.）根据具体的免疫接种计划接种四次。通常，实验组中的小鼠用多肽、pGRA 4、肽/DNA或DNA/肽免疫，对照组中的小鼠用PBS或pEGFP注射。免疫后，IgG抗体和细胞因子的产生水平通过酶联免疫吸附试验（ELISA）测定。用强毒力T.弓形虫RH株结果表明，不同免疫方案（多肽、pGRA 4、肽/DNA、DNA/肽）均能诱导小鼠产生特异性的体液和细胞免疫应答，其中总IgG、IgG 2a亚型和γ-干扰素（IFN-γ）水平较高，提示特异性Th 1免疫被激活。致死攻击后，免疫组小鼠的存活时间（11.8 ± 4.8）d较PBS组和pEGFP组明显延长（P < 0.05）。注射PBS或pEGFP的小鼠在8天内死亡，两组保护水平无明显差异（P > 0.05）。这些结果表明，与其他免疫组相比，编码TgGRA 4的这种DNA初免和肽加强免疫方案可以引发最高水平的体液和细胞免疫应答，这是一种有前途的提高DNA免疫效力的方法。
Toxoplasma gondii is a widespread intracellular parasite, which infects most vertebrate animal hosts and causes zoonotic infection in humans. Vaccine strategy remains a promising method for the prevention and control of toxoplasmosis. T. gondii GRA4 protein has been identified as a potential candidate for vaccine development. In our study, we evaluated the immune response induced by four different immunization vaccination strategies encoding TgGRA4. BALB/c mice were intramuscularly (i.m.) immunized four times according to specific immunization schedules. Generally, mice in experimental groups were immunized with polypeptide, pGRA4, peptide/DNA, or DNA/peptide, and mice in the control groups were injected with PBS or pEGFP. After immunization, the levels of IgG antibodies and cytokine productions were determined by enzyme-linked immunosorbent assays (ELISA). The survival time of mice was also evaluated after challenge infection with the highly virulent T. gondii RH strain. The results showed that mice vaccinated with different immunization regimens (polypeptide, pGRA4, peptide/DNA, or DNA/peptide) elicited specific humoral and cellular responses, with high levels of total IgG, IgG2a isotype and gamma interferon (IFN-γ), which suggested a specific Th1 immunity was activated. After lethal challenge, an increased survival time was observed in immunized mice (11.8 ± 4.8 days) compared to the control groups injected with PBS or pEGFP (P < 0.05). Mice injected with PBS or pEGFP died within 8 days, and there was no significant difference in the protection level in two groups (P > 0.05). These results demonstrated that this DNA prime and peptide boost immunization protocol encoding the TgGRA4 can elicit the highest level of humoral and cellular immune responses compared to other immunized groups, which is a promising approach to increase the efficacy of DNA immunization.
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