口服重组酵母(ApoB100肽段)对小鼠产生特异性抗体的诱导作用

Induction of mice specific antibody by oral administration ofwhole recombinant yeast vaccine(ApoB100)

【目的】开发一种针对低密度脂蛋白成分ApoB100的新型重组酵母疫苗,为动脉粥样硬化(Atherosclerosis,AS)的防治提供一种思路。【方法】利用PCR得到包含编码ApoB100的3 136-3 155位氨基酸残基的目的片段,将其克隆到酵母表达载体JMB88-HA-OVA-MCS上,构建JMB88-HA-OVA-hApoB载体,用硫酸铜诱导表达后获得HA-OVA-hApoB融合蛋白。采用口服的方法对昆明小白鼠免疫成功表达HA-OVA-hApoB蛋白的酵母细胞12周,每周1次,每次1×108个酵母细胞,免疫结束后4周采血,用Western blot检测小鼠是否产生特异性抗体。【结果】PCR扩增获得了859bp的人ApoB100基因。成功构建了JMB88-HA-OVA-hApoB载体。口服免疫12周后,HA-OVA-hApoB免疫组小鼠的血清中含有特异性ApoB100抗体。【结论】灌服表达人ApoB100肽段融合蛋白的重组酿酒酵母能诱导小鼠的免疫反应,使其产生特异性抗体。

[ObjectivelThe objective of this study was to develop a novel recombinant yeast vaccine which targets the low-density lipoprotein apolipoprotein B-100 （ApoB100） to induce a protective immune response,and could improve the prevention and cure of atherosclerosis （AS）. [Method]PCR was used to amplify a target fragment from human genomeeontainng apoB-100, and clone it to express vetor JMB88- HA-OVA-MCS to construct JMB88-HA-OVA-hApoB vector. HA-OVA-hApoB fusion protein was ob- tained after induction using copper sulfate. Oral delivery of 1 X 108 recombinant yeasts with HA-OVA- hApoB protein to each mouse was conducted once a week for 12 weeks. Serum samples of immunized mice were collected 4 weeks after the immunization and analyzed using Western bolt method to check if ApoB100-specific antibody presented. [Result] A 859 bp human ApoBlO0 fragment was obtained by PCR. JMB88-HA-OVA-hApoB vector was established successfully. Oral administration of apoB-100-specific re- combinant yeast vaccine for 12 weeks had a specific immune response to ApoB100 peptide. [Conclusion] O- ral administration of recombinant yeast with ApoB100 fusion protein induced mice to create specific antibody.