恒定链CLIP的2个片段在增强体液免疫效果的比较

发布时间：2019-01-13 08:50

【摘要】：【目的】比较恒定链(invariant chain,Ii)CLIP的2个片段(PBS、GBS)影响免疫载体增强体液免疫的效果。【方法】首先,构建了基于小鼠Ii功能片段与新城疫病毒抗原肽F2的6个嵌合体(Cyt/TM/F2、Cyt/TM/F2/GBS、Cyt/TM/PBS/F2、Cyt/TM/F2/TRIM、Cyt/TM/F2/GBS/TRIM、Cyt/TM/PBS/F2/TRIM);其次,定向克隆至原核表达载体pET-32a中,并转化大肠杆菌Rosetta(DE3)诱导表达和纯化目的蛋白;最后,免疫小鼠,经间接ELISA法测定血清抗体效价,以及比较分析不同嵌合体免疫组免疫效果。【结果】一是所构建的含不同Ii功能片段/F2嵌合体的6个免疫组,均比单独F2的免疫组提高抗体水平1.5-4.9倍;二是在上述6个免疫组中,含CLIP的PBS或GBS的免疫组比不含CLIP的免疫组增强1.6-2.4倍;三是含PBS比含GBS的免疫组增强1.5倍。【结论】Ii胞浆区、跨膜区具有增强免疫效果的作用,而CLIP的PBS在Ii免疫载体中的作用优于GBS。
[Abstract]:[objective] to compare the effects of two fragments (PBS,GBS) of invariant chain,Ii CLIP on the enhancement of humoral immunity by immune vectors. Six chimerism (Cyt/TM/F2,Cyt/TM/F2/GBS,Cyt/TM/PBS/F2,Cyt/TM/F2/TRIM,Cyt/TM/F2/GBS/TRIM,) based on mouse Ii functional fragment and Newcastle disease virus peptide F2 were constructed. Cyt/TM/PBS/F2/TRIM); Secondly, it was cloned into prokaryotic expression vector pET-32a and transformed into Escherichia coli Rosetta (DE3) to induce expression and purification of the target protein. Finally, the antibody titers of serum were determined by indirect ELISA assay, and the immune effects of different chimeric immune groups were compared and analyzed. [results] one of the six immune groups with different Ii functional fragments / F2 chimerism was constructed. The antibody level was 1.5-4.9 times higher than that of F2 immunized group. Second, the PBS or GBS immunized with CLIP was 1.6-2.4 times higher than that without CLIP in the above six immunization groups. [conclusion] the cytoplasmic domain and transmembrane region of Ii have the effect of enhancing immunity, and PBS of CLIP is superior to GBS. in Ii immune vector.
【作者单位】：安徽农业大学安徽省人畜共患病重点实验室;
【基金】：国家自然科学基金(31172306)~~
【分类号】：R392

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