含细粒棘球绦虫Eg95-EgA31融合基因转基因苜蓿免疫Balb/c小鼠脾细胞亚群的变化

Q3 Medicine

中华地方病学杂志 Pub Date : 2010-07-20 DOI:10.3760/CMA.J.ISSN.1000-4955.2010.04.010

Y. Ye, Li Wengui

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Ye, Li Wengui","doi":"10.3760/CMA.J.ISSN.1000-4955.2010.04.010","DOIUrl":null,"url":null,"abstract":"Objective To investigate the change of splenocyte subsets in Balb/c mice immunized with transgenic alfalfa(Medicago sativa)containing Eg95-EgA31 fusion gene of Echinococcus granulosus(Eg) and challenged with Eg protoscoleces.Methods Leaf protein was extracted from transgenic alfalfa containing Eg95-EgA31 fusion gene by heat-coagulation method,and concentration of 20 g/L was used in the study.Meanwhile,leaf protein extracted from the transgenic alfalfa containing blank vector(pBI121)and the normal alfalfa was served as control.Thirty-two female Balb/c mice were randomly divided into 4 groups,8 mice in each group.Oral group was immunized with the leaf protein containing Eg95-EgA31 fusion antigen intragastrically(100μl per mouse);intranasal group was immunized with the leaf protein containing Eg95-EgA31 fusion antigen intranasally(10 μl per mouse);blank vector group was vaccinated intranasally with 10μl leaf protein with blank vector(pBI121);and normal control group was given 100μl normal leaf protein intragastrically.All mice in the above mentioned groups were immunized every 3 days for 2 months.Then,the mice were challenged intraperitoneally with Eg protoscoleces(50 protoscoleces per mouse)8 weeks after last vaccination and sacrified 24 weeks pest infection to separate the splenocytes.Flow cytometry was used to measure the percentages of CD4+ and CD8+ T ceils subsets.Resuits Compared with the normal control group(0.166±0.018,0.083±0.006,2.019 ±0.369),the percentages of CD4+(0.286±0.009)and CD8+(0.102±0.004)T cell subsets and the ratio of CD4+/CD8+(2.814±0.014)in oral group increased significantly (P 0.05).Conclusions CD4+ T cell may play an important role in the protection induced by transgenic alfalfa vaccine against the challenge of Eg protoscoleces.Intragastrical immunization may be a good route. \n \nKey words: \nEchinococcus granulosus; Plants,genetically modified; Medicago sativa; Lymphocyte subsets","PeriodicalId":55880,"journal":{"name":"中华地方病学杂志","volume":"14 1","pages":"387-390"},"PeriodicalIF":0.0000,"publicationDate":"2010-07-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"The change of splenocyte subsets in Balb/c mice by immunization with the transgenic alfalfa(Medicago sativa) containing Eg95-EgA31 fusion gene of Echinococcus granulosus\",\"authors\":\"Y. 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引用次数: 0

摘要

目的观察含有细粒棘球绦虫Eg95-EgA31融合基因的转基因紫花苜蓿免疫Balb/c小鼠后脾细胞亚群的变化。方法采用热凝法提取含有Eg95-EgA31融合基因的转基因紫花苜蓿叶片蛋白，浓度为20 g/L。同时，以含有空白载体(pBI121)的转基因紫花苜蓿叶片蛋白和正常紫花苜蓿叶片蛋白为对照。将32只Balb/c雌性小鼠随机分为4组，每组8只。口服组灌胃含有Eg95-EgA31融合抗原的叶蛋白(100μl /只)，鼻内组灌胃含有Eg95-EgA31融合抗原的叶蛋白(10 μl /只)，空白载体组灌胃含有10μl空白载体(pBI121)的叶蛋白，正常对照组灌胃100μl正常叶蛋白。各组小鼠每3天免疫一次，连续2个月。最后一次接种8周后，腹腔注射Eg原头节(每只50个原头节)，并牺牲24周的感染分离脾细胞。流式细胞术检测CD4+和CD8+ T细胞亚群的百分比。结果与正常对照组(0.166±0.018,0.083±0.006,2.019±0.369)比较，口服组CD4+(0.286±0.009)和CD8+(0.102±0.004)T细胞亚群百分比及CD4+/CD8+比值(2.814±0.014)均显著升高(P < 0.05)。结论CD4+ T细胞可能在转基因苜蓿疫苗对Eg原头节攻击的保护中起重要作用。肠内免疫可能是一个很好的途径。关键词:细粒棘球蚴;转基因植物;紫花苜蓿;淋巴细胞亚群

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The change of splenocyte subsets in Balb/c mice by immunization with the transgenic alfalfa(Medicago sativa) containing Eg95-EgA31 fusion gene of Echinococcus granulosus

Objective To investigate the change of splenocyte subsets in Balb/c mice immunized with transgenic alfalfa(Medicago sativa)containing Eg95-EgA31 fusion gene of Echinococcus granulosus(Eg) and challenged with Eg protoscoleces.Methods Leaf protein was extracted from transgenic alfalfa containing Eg95-EgA31 fusion gene by heat-coagulation method,and concentration of 20 g/L was used in the study.Meanwhile,leaf protein extracted from the transgenic alfalfa containing blank vector(pBI121)and the normal alfalfa was served as control.Thirty-two female Balb/c mice were randomly divided into 4 groups,8 mice in each group.Oral group was immunized with the leaf protein containing Eg95-EgA31 fusion antigen intragastrically(100μl per mouse);intranasal group was immunized with the leaf protein containing Eg95-EgA31 fusion antigen intranasally(10 μl per mouse);blank vector group was vaccinated intranasally with 10μl leaf protein with blank vector(pBI121);and normal control group was given 100μl normal leaf protein intragastrically.All mice in the above mentioned groups were immunized every 3 days for 2 months.Then,the mice were challenged intraperitoneally with Eg protoscoleces(50 protoscoleces per mouse)8 weeks after last vaccination and sacrified 24 weeks pest infection to separate the splenocytes.Flow cytometry was used to measure the percentages of CD4+ and CD8+ T ceils subsets.Resuits Compared with the normal control group(0.166±0.018,0.083±0.006,2.019 ±0.369),the percentages of CD4+(0.286±0.009)and CD8+(0.102±0.004)T cell subsets and the ratio of CD4+/CD8+(2.814±0.014)in oral group increased significantly (P 0.05).Conclusions CD4+ T cell may play an important role in the protection induced by transgenic alfalfa vaccine against the challenge of Eg protoscoleces.Intragastrical immunization may be a good route. Key words: Echinococcus granulosus; Plants,genetically modified; Medicago sativa; Lymphocyte subsets

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来源期刊

中华地方病学杂志我国对人类健康危害特别严重的地方性疾病：克山病、大骨节病、碘缺乏病、地方性氟中毒、地方性砷中毒、鼠疫、布鲁氏菌病、寄生虫、新冠肺炎等疾病，同时还报道多发性自然疫源性疾病。

CiteScore

1.60

自引率

0.00%

发文量

8714

期刊介绍： The Chinese Journal of Endemiology covers predominantly endemic diseases threatening health of the people in the areas affected by the diseases including Keshan disease, Kaschin-Beck Disease, iodine deficiency disorders, endemic fluorosis, endemic arsenism, plague, epidemic hemorrhagic fever, brucellosis, parasite diseases and the diseases related to local natural and socioeconomic conditions; and reports researches in the basic science, etiology, epidemiology, clinical practice, control as well as multidisciplinary studies on the diseases.