Institute of Biomedical Science, National Chung-Hsing University, Taichung, Taiwan.
Division of Immunology and Rheumatology, Department of Internal Medicine, China Medical University Hospital, Taichung, Taiwan.
PLoS One. 2018 Jun 12;13(6):e0198821. doi: 10.1371/journal.pone.0198821. eCollection 2018.
DNA vaccines have recently emerged as a therapeutic agent for treating autoimmune diseases, such as multiple sclerosis. Antiphospholipid antibody syndrome (APS) is an autoimmune disease characterized by β2-glycoprotein I (β2-GPI)-targeting antiphospholipid antibodies (APAs) and vascular thrombosis or obstetrical complications. To examine the therapeutic potential of a β2-GPI DNA vaccine, we administered a vaccine mixed with FK506 as an adjuvant to a mouse model of obstetric APS. First, the pCMV3-β2-GPI DNA vaccine, which encodes the full-length human β2-GPI gene, was constructed. Then, we administered the β2-GPI DNA vaccine in 0.1 ml of saline, mixed with or without 100 μg of FK506, intramuscularly to the mice on days 28, 35 and 42. Blood titers of the anti-β2-GPI antibody, platelet counts, activated partial thromboplastin times (aPTTs), and the percentage of fetal loss were measured. We also stimulated murine splenic T cells ex vivo with β2-GPI and determined the T helper cell proportion and cytokine secretion. The administration of the β2-GPI DNA vaccine mixed with FK506 reduced the blood IgG anti-β2-GPI antibody titers and suppressed APS manifestations in mice. The combination also suppressed interferon-γ and interleukin (IL)-17A secretion but increased the Treg cell proportion and IL-10 secretion in murine splenic T cells following ex vivo stimulation with β2-GPI. Our results demonstrated the therapeutic efficacy of a β2-GPI DNA vaccine and FK506 as an adjuvant in a murine model of obstetric APS. Possible mechanisms include the inhibition of Th1 and Th17 responses and the up-regulation of Treg cells.
DNA 疫苗最近已成为治疗自身免疫性疾病（如多发性硬化症）的一种治疗剂。抗磷脂抗体综合征（APS）是一种自身免疫性疾病，其特征为β2-糖蛋白 I（β2-GPI）靶向抗磷脂抗体（APAs）和血管血栓形成或产科并发症。为了研究β2-GPI DNA 疫苗的治疗潜力，我们将疫苗与 FK506 混合作为佐剂，用于产科 APS 的小鼠模型。首先，构建了编码全长人β2-GPI 基因的 pCMV3-β2-GPI DNA 疫苗。然后，我们在第 28、35 和 42 天，将 0.1ml 生理盐水混合的β2-GPI DNA 疫苗（含或不含 100μg FK506）肌肉内注射到小鼠中。测量抗β2-GPI 抗体的血滴度、血小板计数、活化部分凝血活酶时间（aPTT）和胎儿丢失率。我们还在体外使用β2-GPI 刺激小鼠脾 T 细胞，并确定辅助性 T 细胞比例和细胞因子分泌。β2-GPI DNA 疫苗与 FK506 混合给药可降低血液 IgG 抗β2-GPI 抗体滴度并抑制小鼠 APS 表现。该组合还抑制了干扰素-γ和白细胞介素（IL）-17A 的分泌，但在体外使用β2-GPI 刺激后增加了小鼠脾 T 细胞中的 Treg 细胞比例和 IL-10 分泌。我们的结果表明，β2-GPI DNA 疫苗和 FK506 作为佐剂在产科 APS 的小鼠模型中具有治疗功效。可能的机制包括抑制 Th1 和 Th17 反应以及上调 Treg 细胞。
