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| 1 | Immunogenicity and immunizing protection effect of GAMA gene DNA vaccine on Plasmodium berghei显示文摘Objective: To explore the effect of immunogenicity and immunizing protection of GAMA gene DNA vaccine, which was related with merozoite, ookinete and sporozoite invasion. Methods:Gene fragments were obtained using PCR technique and eukaryotic expression vector(containing immunostimulatory sequence) was built. BALB/c mice were divided into PBS control group, empty vector control group and study group and were immunized at week 0, 3 and 6 respectively. Blood was collected 2 weeks after each immunization and serum was separated to detect the Ig G, Ig G1 and Ig G2 a levels. Spleen of mice was obtained for preparation of splenic mononuclear cell and the cytokine IL-4 and IFN-αlevels were detected. Indirect immunofluorescence and western blot were employed to verify the specificity of antiserum. Sporozoite and merozoite invasion were used respectively to detect the immune protective effect 2 weeks after the third immunization. Ookinete conversion rate in vitro and oocyst numbers of mosquito stomach were observed to evaluate the transmission-blocking levels. Results: In GAMA DNA vaccine group: antiserum could be combined with recombinant protein specifically and green fluorescence signals of merozoite, ookinete and sporozoite were observable, while specific fragments and fluorescence signals were not observable in empty vector group. Compared with control group, specific Ig G in DNA vaccine immunity group significantly increased(P<0.01), and Ig G1 and Ig G2 a all increased(P<0.01). IL-4, IFN-αcontent in study group significantly increased, compared with control group(P<0.01). GAMA DNA vaccine immunity could not obviously block the erythrocyte-stage infection(caused by sporozoite invasion); compared with control group, liver worm load was slightly reduced(P<0.05), and antiserum ookinete numbers(cultured in vitro) had no significant difference with oocyst numbers of mosquito stomach in DNA vaccine group. Conclusions:GAMA has good antigenicity, which could stimulate the body to produce specific immune responses; while DNA vaccine immunity could not play a good protective effect, the effect of which is only limited to the slight reduction of liver worm load, and has no obvious erythrocytestage protective effect and transmission-blocking effect. Therefore, trying other immunization strategies for further research on the value of GAMA(as multi-stage antigen vaccine and multistage combined vaccine components of the life-cycle of plasmodium) is necessary. | Feng Du Si Wang Chen Zhao Ya-Ming Cao En-Jie Luo | 2016 | Asian Pacific Journal of Tropical Medicine2016，9，2： | 1 |
| 2 | 严重发热伴血小板减少综合征病毒核蛋白核酸疫苗的构建及免疫原性研究显示文摘目的 :构建SFTSV的核蛋白核酸疫苗并探讨其免疫原性。方法:采用PCR方法扩增SFTSV核蛋白基因,并用引物引入限制性内切酶酶切位点,克隆入载体p JW4303。经酶切和测序鉴定正确的质粒,用PEI瞬时转染HEK293T细胞,用免疫印迹法鉴定核蛋白的表达。同时以质粒p JW4303作为阴性对照,免疫BALB/c小鼠,酶联免疫法验证其免疫原性。结果:成功构建质粒p JW4303-N,经Western blot证实SFTSV核蛋白能够在HEK293T细胞中表达。ELISA检测免疫后小鼠血清特异性Ig G抗体及其亚型发现,Ig G抗体及其亚型较免疫前明显升高,亚型中以Ig G2a升高为主。结论:SFTSV核蛋白核酸疫苗p JW4303-N具有良好的免疫原性,为进一步研究SFTSV核蛋白奠定了基础。 | 周宜庆 金柯 徐菱遥 李军 | 2015 | 南京医科大学学报(自然科学版)2015，35，7： | 1 |
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