负载热休克蛋白60的致耐受性树突状细胞疫苗对小鼠动脉粥样硬化斑块的影响

Effects of rapamycin-treated HSP60-pulsed dendritic cells on the progression of the atherosclerotic plaque in mice

目的:探讨负载热休克蛋白60(HSP60)的致耐受性树突状细胞(DC)接种对ApoE-/-小鼠动脉粥样硬化斑块的影响。方法:制备ApoE-/-小鼠骨髓DC,分别用HSP60或HSP60加雷帕霉素(rap)处理获得DCHSP60和rap-DCHSP60,体外检测各组DC功能。分别用CFSE染色的DCHSP60和rap-DCHSP60及PBS经静脉接种高脂饲养ApoE-/-小鼠3次,末次接种后8周观察各组斑块面积,CD4+T细胞和DC在斑块中浸润情况及脾细胞对HSP60的反应。结果:体外HSP60负载DC CD86表达显著高于未负载DC,而致耐受DC CD86表达显著低于未负载DC。接种小鼠后,DCHSP60组斑块中炎性细胞浸润明显多于PBS组,DCHSP60组斑块面积大于PBS组;而rap-DCHSP60组炎性细胞浸润明显减少,斑块面积显著小于PBS组;HSP60刺激脾细胞后,DCHSP60组IFN-γ分泌增加,而rap-DCHSP6组IL-10分泌增加,反应呈抗原特异性。结论:负载HSP60的耐受性DC接种可抑制HSP60特异性免疫反应,抑制动脉粥样斑块中的炎症反应和斑块的进展。

AIM： To evaluate whether tolerogenic dendritic cells （DC） loaded with heat shock protein 60 （HSP60） inhibit the progression of aortic atherosclerofic plaque in hypercholesterolemic apolipoprotein E （Apo- E） - null mice. METHODS： Bone marrow derived DC of the mice were loaded with HSP60 and co- cultured with rapamycin to generate tolerogenlc DC. The tolerogenic DC, DC loaded only HSP60 and PBS were injected into the ApoE - null mice at 8 weeks of age for three times at a one - week interval, 8 weeks after the last injection, aorta were harvested for HE staining and anti - CD4 ＋ T cell immunostaining. Responses of plcenic cells to HSP60 were also evaluated. RESULTS： Compared with DC, DCHSP60 expressed higher levels of CD86, and stimulated T lymphocytes to proliferation significantly, while the tolerogenic DC expressed lower levels of CD86, and inhibited T lymphocytes to proliferation. After immunization with different injection, the numbers of CD4^＋ T cells in plaque were increased significantly in DCHSP60 group vs in PBS group （ P 〈 0.01 ）. On the other hand, they were reduced significantly in rap- DCHSP60 group vs in PBS group （ P 〈 0.01）. Plaque axeas in the tolerogenic DC group were smaller than that in the PBS group （ P 〈 0.01）. Stimulated by HSP60, plcenic cells in tolerogenic DC group secreted more IL- 10, while in DCHSP60 group more IFN- T secretion was observed. CONCLUSION： HSP60 specific tolerogenic DC immunization attenuated the progression of plaque, indicating a new immune therapy for atherosclerosis.