Departamento de Bioquímica y Biología Estructural, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Ciudad de México, México.
Departamento de Bioquímica y Biología Estructural, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Ciudad de México, México; Max Delbrück-Centrum für Molekulare Medizin, Berlin, Germany.
Arch Med Res. 2018 Oct;49(7):456-470. doi: 10.1016/j.arcmed.2019.01.007. Epub 2019 Feb 18.
Atherosclerosis as an inflammatory disease involved in the etiology of cardiovascular disease worldwide, in our days demands an array of different therapeutic approaches in order to soon be able to visualize an effective prevention. Based on an immunotherapeutic approach, we designed a non-invasive vaccine (HB-ATV-8), contained in a micellar nanoparticle composed of lipids and a peptide segment derived from the C-terminus of the cholesterol-ester transfer protein (CETP). Now we extend our successful proof of concept from the rabbit to a porcine model and investigated its effect in an attempt to undoubtedly establish the efficacy of vaccination in a model closer to the human.
A preclinical trial was designed to study the efficacy of vaccine HB-ATV-8 in pigs (Large White × Landrace). Male experimental animals were fed with standard diet (control), high fat diet (HFD) or the same HFD but treated with HB-ATV-8 (HFD + Vaccine) applied nasally for up to 7 months. All biochemical and enzymatic analyses were performed in peripheral venous blood and thoracic aorta and liver samples examined using conventional, two-photon excitation and second harmonic generation microscopy to identify atherosclerotic and hepatic lesions. mRNA concentrations for KLF2, ACTA2, SOD1, COL1A1 genes and protein levels for PPARα and ABCA1 were quantified in aorta and liver respectively using qPCR and Western blot analysis.
The administration of vaccine HB-ATV-8 induced anti-CETP IgG antibodies and reduced atherosclerotic and hepatic lesions promoted by the high fat diet. In addition, plasma triglyceride levels of vaccine treated pigs fed the HFD were similar to those of control group, in contrast to high concentrations reached with animals exclusively fed with HFD. Moreover, HFD promotes a tendency to decrease hepatic PPARα levels and increase in aorta gene expression of KLF2, ACTA2, SOD1 and COL1A1, while vaccine application promotes recovery close to control values.
Vaccine HB-ATV-8 administration constitutes a promissory preventive approach useful in the control of atherogenesis and fatty liver disease. The positive results obtained, the non-invasive characteristics of the vaccine, the simple design employed in its conception and its low production cost, support the novelty of this therapeutic strategy designed to prevent the process of atherogenesis and control the development of fatty liver disease.
动脉粥样硬化作为一种炎症性疾病，参与了全球心血管疾病的病因，如今需要多种不同的治疗方法，以便尽快能够看到有效的预防效果。基于免疫治疗方法，我们设计了一种非侵入性疫苗（HB-ATV-8），包含在由脂质和胆固醇酯转移蛋白（CETP）C 端衍生肽段组成的胶束纳米颗粒中。现在，我们将从兔子身上获得的成功概念验证扩展到猪模型中，并研究其效果，试图在更接近人类的模型中确定疫苗接种的疗效。
设计了一项临床前试验，以研究疫苗 HB-ATV-8 在猪（大白 × 长白）中的疗效。雄性实验动物用标准饮食（对照）、高脂肪饮食（HFD）或相同的 HFD 但用 HB-ATV-8 治疗（HFD+疫苗）喂养长达 7 个月。所有生化和酶分析均在外周静脉血中进行，通过常规、双光子激发和二次谐波产生显微镜检查胸主动脉和肝组织样本，以识别动脉粥样硬化和肝损伤。使用 qPCR 和 Western blot 分析分别定量主动脉和肝中 KLF2、ACTA2、SOD1、COL1A1 基因的 mRNA 浓度和 PPARα和 ABCA1 的蛋白水平。
疫苗 HB-ATV-8 的给药诱导了抗 CETP IgG 抗体，并减少了高脂肪饮食引起的动脉粥样硬化和肝损伤。此外，用 HFD 喂养的疫苗处理猪的血浆甘油三酯水平与对照组相似，而仅用 HFD 喂养的动物则达到了较高的浓度。此外，HFD 促进了肝中 PPARα 水平降低和主动脉中 KLF2、ACTA2、SOD1 和 COL1A1 基因表达增加的趋势，而疫苗的应用则促进了接近对照值的恢复。
疫苗 HB-ATV-8 的给药构成了一种有前途的预防方法，可用于控制动脉粥样硬化和脂肪肝疾病。所获得的积极结果、疫苗的非侵入性特征、其设计中采用的简单设计及其低廉的生产成本，支持了这种旨在预防动脉粥样硬化过程和控制脂肪肝疾病发展的治疗策略的新颖性。
