Xiong Qiyan, Feng Jiao, Zhang Yu, Sun Yunxiao, Lu Yong, Li Taiming, Zhang Xiaohong, Cao Rongyue, Jin Liang, Wu Jie
Minigene Pharmacy Laboratory, School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China; Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture, National Center for Engineering Research of Veterinary Bio-products, Nanjing 210014, China.
Minigene Pharmacy Laboratory, School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China; Department of Biological Engineering, Liaoning Economic Management Cadre Institute, Shenyang 110122, China.
Immunol Lett. 2016 Feb;170:80-7. doi: 10.1016/j.imlet.2015.12.006. Epub 2015 Dec 28.
Previous evidence has proved the ability of immunization with heat shock protein (HSP) 60/65 to induce atherosclerosis. P277, a 24-residue peptide of human HSP60, is a promising peptide vaccine against autoimmune diabetes. But as a fragment of HSP60, its potential ability of promoting atherosclerosis has never been investigated yet. In the present study, the rabbits fed with normal standard diet or high cholesterol diet were immunized with P277 or PBS emulsified in incomplete Freund's adjuvant 4 times at 4-week intervals. Atherosclerotic lesions of the rabbits receiving P277 treatment and fed with high cholesterol diet increased significantly compared with those of the rabbits receiving PBS treatment and the same diet. However, no obvious lesions were found in the two groups of rabbits fed with the normal standard diet. Significant expression of P277 was detected in the high cholesterol diet-induced atherosclerotic lesions and heat-stressed endothelial cells. Surface exposure of P277 was also observed in the stressed cells. In the subsequent assay of endothelial cells in vitro, the purified anti-P277 antibodies mediated a noticeable cytotoxicity to the stressed cells with the participation of complement. In conclusion, subcutaneous immunization with P277 emulsified in IFA can aggravate the atherosclerosis in high cholesterol diet-fed rabbits. Surface expression of P277 was observed on stressed endothelial cells, and were suggested to mediate the autoimmune attack and promote the disease.
先前的证据已证明用热休克蛋白（HSP）60/65进行免疫可诱发动脉粥样硬化。P277是人类HSP60的一个24个残基的肽段，是一种有前景的抗自身免疫性糖尿病的肽疫苗。但作为HSP60的一个片段，其促进动脉粥样硬化的潜在能力尚未得到研究。在本研究中，给喂食正常标准饮食或高胆固醇饮食的兔子每隔4周用不完全弗氏佐剂乳化的P277或PBS免疫4次。与接受PBS处理并喂食相同饮食的兔子相比，接受P277处理并喂食高胆固醇饮食的兔子的动脉粥样硬化病变显著增加。然而，在两组喂食正常标准饮食的兔子中未发现明显病变。在高胆固醇饮食诱导的动脉粥样硬化病变和热应激内皮细胞中检测到P277的显著表达。在应激细胞中也观察到P277的表面暴露。在随后的体外内皮细胞测定中，纯化的抗P277抗体在补体的参与下介导了对应激细胞的明显细胞毒性。总之，用不完全弗氏佐剂乳化的P277皮下免疫可加重高胆固醇饮食喂养兔子的动脉粥样硬化。在应激内皮细胞上观察到P277的表面表达，提示其介导自身免疫攻击并促进疾病发展。
