霍乱毒素B亚单位与热休克蛋白65功能表位融合蛋白皮下免疫加剧动脉粥样硬化  

Aggravating Atherosclerosis of the Fusion-Protein of CTB and Heat Shock 65 Epitopes by Subcutaneous Injection

作　　者：孙云霄[1,2,3] 张礼标[1,2] 王华倩[3,4] 张宇[3] 熊祺琰[3] 李泰明[3] 曹荣月[3] 

机构地区：[1]广东省昆虫研究所 [2]华南濒危动物研究所,广东省广州市510260 [3]中国药科大学生命科学与技术学院微基因药物实验室,江苏省南京市210009 [4]广东工业大学药学院,广东省广州市510260

出　　处：《中国动脉硬化杂志》2014年第10期988-992,共5页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金资助项目(81102358);广东省科学院优秀青年人才基金(rcjj201104);广东省自然科学基金(S2012040007488)

摘　　要：目的研究热休克蛋白65 B细胞表位经皮下免疫高脂膳食动物对动脉粥样硬化(As)斑块生成的影响。方法利用HSP65上与炎症性自身免疫性疾病相关的两段功能表位P1(179-190)、P2(31-46),以CTB为载体蛋白构建高效表达载体p ET28a-CTB-p117,提取纯化获得可用于动物实验的蛋白,皮下免疫新西兰大白兔,比较抗HSP65抗体、血液总胆固醇含量变化与主动脉粥样硬化斑块的生成情况。结果能够诱导产生抗HSP65的抗体,血液中总胆固醇含量与对照组比较显著提高,并且能显著增加主动脉As斑块生成(P<0.05)。结论 HSP65相关表位蛋白以CTB为载体蛋白,在弗氏不完全佐剂条件下,经皮下免疫能显著加重As斑块生成,分析是因为抗HSP65抗体生成,促进炎症反应,脂类代谢变化进而促进As斑块的形成。可基于此,利用以上表位,改变免疫方式,设计出可用于降低斑块生成的抑制As斑块生成的免疫调节剂。对于CTB单独皮下免疫也致As斑块加重的结果需进一步实验考察验证。Aim To study the immunization effects of B epitopes of heat shock protein 65 on the atherosclerosis plaque formation of high fat diets animals. Methods By using two functional epitopes P1（ 179-190）,P2（ 31-46） of HSP65 related to inflammatory autoimmune disease,high-level expression vector of p ET28a-CTB-p117 was built with CTB as a carrier protein. The fusion protein of CTB-p117 was acquired by expression and purification process and used to immunize the animals by subcutaneous and intracutaneous injections. Results Anti-HSP65 antibodies were induced and both blood total cholesterol levels and aortic As plaques significantly increased compared with control group（ P 〈 0. 05）.Conclusions The HSP65 epitopes（ CTB as the carrier protein） were able to aggravate As plaque formation after subcutaneous injections which could be the reasons that the induced anti-HSP65 antibodies promoted inflammatory reaction,lipid metabolic changes,and then accelerate the As plaque formation. The above HSP65 epitopes can be used to construct the anti-As vaccine to inhibit the As plaque formation by different immunization way.

关 键 词：动脉粥样硬化 热休克蛋白 免疫 

分 类 号：R9[医药卫生—药学]