Objective To construct the eukaryotic expression recombinant plasmid, pcIFN γ, as a genetic adjuvant and observe the immune responses elicited by pcDNA3 rhoptry protein 1 (pc ROP1) combined with pcIFN γ aga...Objective To construct the eukaryotic expression recombinant plasmid, pcIFN γ, as a genetic adjuvant and observe the immune responses elicited by pcDNA3 rhoptry protein 1 (pc ROP1) combined with pcIFN γ against Toxoplasma gondii (T gondii) infection in mice Methods A fragment of the IFN γ gene was directly inserted into the pcDNA3 plasmid and identified by two restriction endonucleases digestion pcIFN and pcROP1 DNA was injected into the left leg muscle of mice at a dosage of 100?μg, and a booster vaccination was given at the same dosage after two weeks Control groups were injected with pcDNA3 blank plasmid or normal saline At 30, 50 and 70 days after booster injection, kinetic tests were carried out: MTT assay for the proliferation response of T lymphocyte cells and the activity of NK cells, sandwich ABC ELISA for the determination of IFN γ, IL 2 and IL 10; a serum enzymetic aassay for nitric oxide (NO) in sera and ELISA for the titer of IgG antibody in sera Results The recombinant plasmid, pcIFN γ was constructed The proliferation response of spleen T lymph cells, NK cell killing activity, and serum levels of IFN γ, IL 2 and NO in mice injected with pcROP1 and pcIFN γ were higher than in those injected with pcROP1 alone There was no difference in IgG antibody levels between the two groups Conclusion The genetic adjuvant, pcIFN γ, could enhance the cellular immune response induced by DNA vaccine of pcROP1 in mice against Toxoplasma gondii infection展开更多
目的 研究日本血吸虫卵黄铁蛋白基因 DNA免疫小鼠诱导的保护性免疫力。方法 将构建的日本血吸虫卵黄铁蛋白基因真核表达重组质粒 p L XSN- Fer l经左后腿肌肉注射 BAL B/ c小鼠 ,共免疫 2次 ,间隔 2周。末次免疫后 5周以血吸虫尾蚴攻...目的 研究日本血吸虫卵黄铁蛋白基因 DNA免疫小鼠诱导的保护性免疫力。方法 将构建的日本血吸虫卵黄铁蛋白基因真核表达重组质粒 p L XSN- Fer l经左后腿肌肉注射 BAL B/ c小鼠 ,共免疫 2次 ,间隔 2周。末次免疫后 5周以血吸虫尾蚴攻击感染 ,攻击感染后 6周计数成虫负荷及肝组织虫卵数和死亡卵百分比。设空质粒免疫组为对照组。结果 p L XSN- Fer l免疫小鼠 ,诱生的Ig G亚类主要是 Ig G2 a和 Ig G2 b;免疫组可产生较高水平的 Ig A和 IFN- γ应答。 p L XSN- Fer l免疫小鼠诱生一定程度的减虫率 34 .0 8% ,减卵率 36 .83% ,死亡卵百分比为 30 .13%。结论 p L XSN-Fer l免疫小鼠能产生较明显的保护性免疫力 ,值得进一步深入研究。展开更多
文摘Objective To construct the eukaryotic expression recombinant plasmid, pcIFN γ, as a genetic adjuvant and observe the immune responses elicited by pcDNA3 rhoptry protein 1 (pc ROP1) combined with pcIFN γ against Toxoplasma gondii (T gondii) infection in mice Methods A fragment of the IFN γ gene was directly inserted into the pcDNA3 plasmid and identified by two restriction endonucleases digestion pcIFN and pcROP1 DNA was injected into the left leg muscle of mice at a dosage of 100?μg, and a booster vaccination was given at the same dosage after two weeks Control groups were injected with pcDNA3 blank plasmid or normal saline At 30, 50 and 70 days after booster injection, kinetic tests were carried out: MTT assay for the proliferation response of T lymphocyte cells and the activity of NK cells, sandwich ABC ELISA for the determination of IFN γ, IL 2 and IL 10; a serum enzymetic aassay for nitric oxide (NO) in sera and ELISA for the titer of IgG antibody in sera Results The recombinant plasmid, pcIFN γ was constructed The proliferation response of spleen T lymph cells, NK cell killing activity, and serum levels of IFN γ, IL 2 and NO in mice injected with pcROP1 and pcIFN γ were higher than in those injected with pcROP1 alone There was no difference in IgG antibody levels between the two groups Conclusion The genetic adjuvant, pcIFN γ, could enhance the cellular immune response induced by DNA vaccine of pcROP1 in mice against Toxoplasma gondii infection
文摘目的 研究日本血吸虫卵黄铁蛋白基因 DNA免疫小鼠诱导的保护性免疫力。方法 将构建的日本血吸虫卵黄铁蛋白基因真核表达重组质粒 p L XSN- Fer l经左后腿肌肉注射 BAL B/ c小鼠 ,共免疫 2次 ,间隔 2周。末次免疫后 5周以血吸虫尾蚴攻击感染 ,攻击感染后 6周计数成虫负荷及肝组织虫卵数和死亡卵百分比。设空质粒免疫组为对照组。结果 p L XSN- Fer l免疫小鼠 ,诱生的Ig G亚类主要是 Ig G2 a和 Ig G2 b;免疫组可产生较高水平的 Ig A和 IFN- γ应答。 p L XSN- Fer l免疫小鼠诱生一定程度的减虫率 34 .0 8% ,减卵率 36 .83% ,死亡卵百分比为 30 .13%。结论 p L XSN-Fer l免疫小鼠能产生较明显的保护性免疫力 ,值得进一步深入研究。
