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SAG1与IL-2基因佐剂联合DNA介导免疫对小鼠弓形虫感染的保护作用

Protective effect of DNA-mediated immunization with a combination of SAG1 and IL-2 gene adjuvant against infection of Toxoplasma gondii in mice.

作者信息

Chen Guanjin, Chen Haifeng, Guo Hong, Zheng Huanqin

机构信息

Institute of Parasitology, Sun Yat-Sen University of Medical Sciences, Guangzhou 510089, China.

出版信息

Chin Med J (Engl). 2002 Oct;115(10):1448-52.

Abstract

OBJECTIVE

To characterize the immune response induced by SAG1 encoding plasmid combined with IL-2 gene adjuvant in mice and to assess the protective effect of this vaccination against toxoplasmosis.

METHODS

Mice were co-injected intramuscularly with plasmid encoding Toxoplasma gondii SAG1 plus murine IL-2 expression vector at a dose of 100 microg. Booster immunizations were employed 2 more times at 3-week interval. As controls, mice were inoculated with PBS or empty plasmid pcDNA3. Humoral and cellular responses were assayed using ELISA for the determination of Ab, Ab isotype and IFN-gamma, as well as IL-4. To detect the integration and dissemination of DNA in the injected mice, PCR and in situ hybridization were performed. All mice were then infected with highly virulent RH tachyzoites of Toxoplasma gondii intraperitoneally.

RESULTS

Significant increases in specific IgG levels were observed in mice after immunization three times with SAG1 expression plasmid. With respect to the IgG isotype, co-inoculation of IL-2 expression plasmid enhanced the level of IgG2a and the production of IFN-gamma. Challenging mice by vaccinating with combined plasmids with RH tachyzoites resulted in prolonged survival.

CONCLUSION

Humoral and cytokine responses elicited by SAG1 DNA immunization can be modulated by co-inoculation with IL-2 expression plasmid. The use of DNA vaccine in combination with an appropriate cytokine gene to prevent T. gondii infection warrants further investigation.

摘要

目的

鉴定编码SAG1的质粒与白细胞介素-2(IL-2)基因佐剂联合诱导的小鼠免疫反应,并评估该疫苗接种对弓形虫病的保护作用。

方法

将编码刚地弓形虫SAG1的质粒与小鼠IL-2表达载体以100微克的剂量联合肌肉注射给小鼠。每隔3周再进行2次加强免疫。作为对照,小鼠接种磷酸盐缓冲液(PBS)或空质粒pcDNA3。使用酶联免疫吸附测定法(ELISA)检测体液和细胞反应,以测定抗体(Ab)、抗体亚型以及干扰素-γ(IFN-γ)和白细胞介素-4(IL-4)。为检测DNA在注射小鼠体内的整合和扩散情况,进行了聚合酶链反应(PCR)和原位杂交。然后所有小鼠经腹腔注射高毒力的刚地弓形虫RH速殖子。

结果

用SAG1表达质粒免疫小鼠3次后,观察到特异性IgG水平显著升高。关于IgG亚型,共接种IL-2表达质粒提高了IgG2a水平和IFN-γ的产生。用联合质粒疫苗接种的小鼠再感染RH速殖子后存活时间延长。

结论

与IL-2表达质粒共接种可调节SAG1 DNA免疫引发的体液和细胞因子反应。使用DNA疫苗联合适当的细胞因子基因预防刚地弓形虫感染值得进一步研究。

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