摘要
目的对pSG.SS.Flk-1265-2493.C3d3.YL及pSG.SS.Flk-1265-2493.YL真核表达质粒的免疫效应进行初步研究。方法构建pSG.SS.Flk-1265-2493.C3d3.YL及pSG.SS.Flk-1265-2493.YL真核表达质粒后,48只BALB/c小鼠随机均分为疫苗组、疫苗对照组、质粒对照组、空白对照组(12只/组),分别肌肉注射接种此2种质粒、pSG.SS.YL质粒和PBS。免疫后动物:①每组6只取脾脏分离细胞,MTT法测定经Flk-1蛋白刺激后的增殖效应,乳酸脱氢酶释放法测定对培养小鼠肺微血管内皮细胞的杀伤作用;②其余6只动物于不同时间点断尾取血分离血清,ELISA法测定抗Flk-1抗体水平的动态变化。结果经Flk-1蛋白刺激,疫苗组小鼠脾淋巴细胞的增殖效应明显较强,刺激指数显著高于其他3组(P均<0.01),对靶细胞(MVECs)的杀伤活性在不同效/靶比时均显著高于其他3组(P均<0.01)。疫苗组动物血清抗Flk-1抗体比对照疫苗组抗体阳性反应出现早2周,二者峰值效价均出现在5周(血清稀释度:1∶3840vs1∶60),前者12周时仍保持较高水平(1∶960),后者10周时抗体阳性反应已消失。结论C3d3的连接有效增强了Flk-1265-2493的免疫原性。
Objective To evaluate the immunological effect of eukaryotic expression plasmids pSG. SS. Flk-1265-2493. C3d3. YL and pSG. SS. Flk-1265-2493. YL. Mothods After construction of eukaryotic expression plasmids pSG. SS. Flk-1265-2493. C3d3. YL and pSG. SS. Flk-1265-2493. YL, BALB/c female mice （n =48） were separated into four groups （n = 12 per group） and were inoculated by intramuscular injection of the two plasraids, pSG. SS. YL plasmid （plasmid control） and PBS （blank control）, respectively. After inoculation, proliferation of spleen lymphocytes stimulated with Flk-1 protein was studied using MTT method and the specific cytotoxieity T lymphocyte （CTL） effect on target cells （microvaseular endothelial cells, MVECs） was assayed with lactate dehydrogenase-release method. Then anti-Flk-1 antibody titers in the immune sera were assayed with indirect enzyme-linked immunosordent assay （ELISA）. Results After stimulation with Flk-1 antigen, the proliferation and specific CTL activities of spleen lymphocytes in mice immunized with pSG. SS. Flk-1265-493. C3d3. YL were obviously stronger than those in other three groups. Although both plasmids could elicit Flk-1 specific antibody, in pSG. SS. Flk-1265-2493.C3d3. YL immunized mice, the antibody presented 2 weeks earlier, and the titer was significantly higher than that in mice immunized with pSG. SS. Flk-1265-2493. YL. Conclusion The results show that the C3d3 fusion strategy can enhance the immunogenicity of Flk-1 significantly
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2009年第13期1239-1242,共4页
Journal of Third Military Medical University
基金
国家自然科学基金(30571864)~~
