论文部分内容阅读
目的:通过分枝杆菌热休克蛋白保守性B细胞表位多次免疫小鼠,研究热休克蛋白65(HSP65)与动脉粥样硬化之间的关系。方法:PCR技术扩增出HSP65上与动脉粥样硬化密切相关的6个B细胞表位基因。将这6个表位的基因分成两组,分别与CTB基因相融合并在大肠杆菌中表达。两种蛋白表达产物经过分离、纯化、混合复性,复性后的蛋白经GM1-ELISA证明可在体外自组装成5聚体。用复性后的重组蛋白小剂量多次滴鼻免疫ICR小鼠,然后用大量的HSP65蛋白经皮下免疫,采集血样,ELISA检测小鼠血清中抗HSP65的抗体。结果:重组蛋白预免疫ICR小鼠后,可以使免疫组较对照组对随后的大量HSP65刺激产生更加强烈的免疫反应。结论:分枝杆菌热休克蛋白保守性B细胞表位经鼻黏膜的反复预免疫强化了小鼠对随后攻击性免疫的反应。
OBJECTIVE: To study the relationship between heat shock protein 65 (HSP65) and atherosclerosis by immunizing mice with mycobacterial heat shock protein conserved B cell epitopes multiple times. Methods: Six B cell epitope genes closely related to atherosclerosis on HSP65 were amplified by PCR. The six epitopes were divided into two groups and fused with CTB gene respectively and expressed in E. coli. After the two protein expression products were separated, purified and mixed refolded, the renatured protein was proved to be self-assembled into 5-mer in vitro by GM1-ELISA. The ICR mice were immunized with small amount of refolded recombinant protein and then subcutaneously immunized with a large amount of HSP65 protein. The blood samples were collected and the anti-HSP65 antibody in serum was detected by ELISA. Results: After pre-immunization of ICR mice with recombinant protein, the immunized group could produce a more intense immune response to the subsequent massive HSP65 stimulation than the control group. Conclusions: The repeated pre-immunization of mycobacterial heat shock protein-conserved B cell epitopes through the nasal mucosa potentiates the mouse response to subsequent offensive immunity.