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细粒棘球蚴重组抗原蛋白BCG-egG1Y162免疫原性的初步研究 预览

Immunogenicity analysis of recombinant protein BCG-egG1Y162 of Echinococcus granulosus
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摘要 目的初步研究细粒棘球蚴重组抗原蛋白BCG-egG1Y162的免疫原性。方法选取60只小鼠分别分为3组实验组、重组卡介苗组和正常组小鼠分别注射重组抗原蛋白BCG-egG1Y162、重组卡介苗(BCG)和生理盐水(NS)。在免疫的第0、2、4、6、8周收集各组小鼠的血清,通过酶联免疫吸附法(ELISA法)检测血清中抗体IgG的动态变化水平。在第8周随机处死各组一半数量的小鼠提取脾淋巴细胞,分别通过四甲基偶氮咗蓝实验(MTT法)以及ELISA法检测小鼠脾淋巴细胞体外增殖反应和细胞上清中IFN-γ、IL-4的水平变化。在免疫后第8周用活棘球蚴原头蚴感染各组存活的小鼠,感染后第24周处死各组小鼠,检测血清中IgG水平、包囊个数及其直径。结果(1)免疫后的第2~8周,BCG-egG1Y162免疫组小鼠血清中IgG持续升高,与正常组和BCG组相比差异具有统计学意义(P<0.05)。BCG组和正常组的IgG水平之间的差异不具有统计学意义(P>0.05)。(2)BCG-egG1Y162免疫组小鼠的脾淋巴细胞在体外能被BCG-egG1Y162特异性的刺激增生,增殖水平显著高于BCG组和正常组(P<0.05),其上清中IFN-γ、IL-4的水平以及IFN-γ/IL-4的比值同样显著高于正常组和BCG组(P<0.05)。这些指标在BCG组和正常组之间的差异不具有统计学意义(P>0.05)。(3)在各组小鼠感染活棘球蚴原头蚴后的第24周,BCG-egG1Y162免疫组具有87.3%的免疫保护力,并且血清中的IgG水平显著高于BCG组和正常组(P<0.05)。结论细粒棘球蚴重组抗原蛋白BCG-egG1Y162免疫小鼠后可上调小鼠血清中抗体IgG的水平,刺激脾淋巴细胞增殖活化,特异性地增强脾淋巴细胞的Th1免疫反应和机体抵抗细粒棘球蚴感染的能力,说明BCG-egG1Y162是具有研究价值的细粒棘球蚴病候选疫苗分子。 Objective To investigate the immunogenicity of recombinant protein BCG-egG1 Y162 of Echinococcus granulosus. Methods 60 mice were divided into three groups: the experimental group(BCG-egG1 Y162 immunization group), BCG group and normal saline group(NS group). The mice were injected with BCG-egG1 Y162, BCG and saline,respectively. On 2 nd to 8 th weeks, serum IgG levels of three groups were examined by ELISA. On 8 th week, the splenic T lymphocytes were isolated for the half total number on mice. The proliferation of splenic T lymphocytes and the levels of IFN-γand IL-4 in the culture supernatant were measured by MTT and ELISA, respectively. On 8 th week,mice challenged with E. granulosus protoscoleces. On 24 th week after infection,the mice were killed. The level of IgG and the number of the fresh hydatid cysts were evaluated. Results(1) The level of IgG in mice immunized with BCG-egG1 Y162 increased obviously on 2 nd to 8 th weeks, and the difference was significant compared to NS group and BCG group(P<0.05). There were no significant difference between the level of IgG in NS group and BCG group(P>0.05).(2) The spleen lymphocyte proliferation of BCG-egG1 Y162 immunized mice was stimulated by BCG-egG1 Y162 in vitro. The proliferation level of spleen lymphocyte was significantly higher than that of BCG group and normal group(P<0.05). The levels of IFN-gamma and IL-4 in supernatant and the ratio of IFN-gamma/IL-4 were also significantly higher than that of normal group and BCG group(P<0.05). There was no significant difference in these indexes between BCG group and normal group(P>0.05).(3) On 24 th week after infection, the number and the size of fresh hydatid cyts in BCG-egG1 Y162 immunization group was significant less than the normal group with a protective efficacy of 87.3%. Serum IgG levels in BCG-egG1 Y162 immunization group was significantly higher than that of NS group and BCG group. Conclusion These data indicated that BCG-egG1 Y162 protein immunization could induce the production of serum IgG, meanwhile
作者 李海丽 刘永太 张峰波 李玉娇 李智伟 赵骁 丁剑冰 王红英 LI Haili;LIU Yongtai;ZHANG Fengbo;LI Yujiao;LI Zhiwei;ZHAO Xiao;DING Jianbing;WANG Hongying(First Clinical Medical College, Xinjiang Medical University, Urumqi 830011, China;School of Preclinical Medicine, Xinjiang Medical University, Urumqi 830011, China;Department of Clinical Laboratory, the First Division Hospital of Xinjiang Production and Construction Crops, Aksu Xinjiang 843000, China;State Key Laboratory of Pathogenesis, Prevention, Treatment of Central Asian High Incidence Diseases, Xinjiang Medical University, Urumqi 830011, China)
出处 《新疆医科大学学报》 CAS 2019年第5期593-598,共6页 Journal of Xinjiang Medical University
基金 新疆维吾尔自治区自然科学基金医学联合基金项目(2014211C006).
关键词 细粒棘球蚴病 免疫原性 重组蛋白BCG-egG1Y162 Th1免疫反应 Echinococcus granulosus immunogenicity antigen BCG-egG1Y162 Th1 immune response
作者简介 李海丽(1988-),女,在读硕士,初级技师,研究方向:寄生虫免疫;通信作者:王红英,女,教授,硕士生导师,研究方向:感染免疫,E-mail:736932811@qq.com.
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