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[用重组肌动蛋白疫苗免疫ICR小鼠对细粒棘球绦虫免疫反应的研究]

[Study on immune response in ICR mice by immunization with recombinant myophilin vaccine against Echinococcus granulosus].

作者信息

Ma Rui, Shi Zhi-Yun, Wang Ya-Na, Li Zong-Ji, Sun Jun-Feng, Zhao Wei

机构信息

Higher Health Vocational and Technical College of Ningxia Medical University, Yinchuan 750004, China.

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2012 Oct 30;30(5):357-60.

Abstract

OBJECTIVE

To investigate the immune response and the protection in mice induced by the recombinant myophilin protein of Echinococcus granulosus.

METHODS

Thirty-six male ICR mice of 6-8 weeks old were randomly divided into groups A, B and C each with 12. The mice in the 3 groups were subcutaneously immunized with Eg myophilin protein, blank plasmid protein or PBS, respectively, by 3 times and challenged with protoscoleces of E. granulosus 2wk after the last vaccination. Mice were sacrificed 20wk after the infection, the hydatid cysts were collected for measuring the weight reduction. Spleens were obtained and the splenocytes were separated and cultured in vitro with EgAg or ConA stimulus for 4-5 h. The subsets of CD4+ and CD8+ T cells were measured by FACsort. The proliferation of splenocytes was determined by MTT method with blank plasmid and PBS as control.

RESULTS

The average weight of the hydatid cysts in the immunized group decreased by 69.1% in comparison to the blank plasmid and PBS groups. The CD4+ subset [(29.7 +/- 0.9)%] and CD84+ subset [(9.7 +/- 0.8)%] in group A increased significantly than group C, [(11.6 +/- 1.4)%] and [(7.8 +/- 0.2)%] respectively (P < 0.01 or < 0.05). The ratio of CD4+/CD8+ subsets in group A (3.061 +/- 0.015) was also higher than group C (1.487 +/- 0.106) (P < 0.01). Without stimulation, the proliferation of T lymphocytes in group A(0.237 +/- 0.009) was higher than group C (0.159 +/- 0.005) (P < 0.01), with EgAg or ConA stimulus, it was also higher in group A than that of group C (P < 0.01).

CONCLUSION

The recombinant myophilin protein of E. granulosus can induce the proliferation of splenocytes and Th1 response in mice, and the CD4+ T cells subset may bear a part in the induced protection against the challenge of protoscoleces.

摘要

目的

研究细粒棘球绦虫重组亲肌球蛋白蛋白诱导小鼠的免疫反应及保护作用。

方法

将36只6 - 8周龄雄性ICR小鼠随机分为A、B、C组,每组12只。3组小鼠分别皮下免疫细粒棘球绦虫亲肌球蛋白蛋白、空质粒蛋白或PBS,共免疫3次,末次免疫后2周用细粒棘球绦虫原头蚴攻击。感染后20周处死小鼠,收集包虫囊肿测量重量减轻情况。取脾脏分离脾细胞,体外分别用细粒棘球绦虫抗原(EgAg)或刀豆蛋白A(ConA)刺激培养4 - 5小时。用流式细胞仪检测CD4⁺和CD8⁺T细胞亚群。以空质粒和PBS为对照,采用MTT法检测脾细胞增殖情况。

结果

免疫组包虫囊肿平均重量较空质粒组和PBS组减轻69.1%。A组CD4⁺亚群[(29.7±0.9)%]和CD8⁺亚群[(9.7±0.8)%]较C组[分别为(11.6±1.4)%和(7.8±0.2)%]显著增加(P<0.01或<0.05)。A组CD4⁺/CD8⁺亚群比值(3.061±0.015)也高于C组(1.487±0.106)(P<0.01)。未刺激时,A组T淋巴细胞增殖率(0.237±0.009)高于C组(0.159±0.005)(P<0.01),经EgAg或ConA刺激后,A组仍高于C组(P<0.01)。

结论

细粒棘球绦虫重组亲肌球蛋白蛋白可诱导小鼠脾细胞增殖及Th1反应,CD4⁺T细胞亚群可能参与诱导的针对原头蚴攻击的保护作用。

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