抗磷脂综合征(APS)是一种自身免疫性疾病,其特征是产生β2-糖蛋白I(β2GPI)依赖性自身抗体,血管血栓形成或产科并发症。大约20%的APS患者对目前的治疗是难以治疗的。Crassolide,一种从软珊瑚中提取的类胶膜二萜,是一个潜在的治疗候选。这里,为了检查克拉索利的抗炎特性,我们首先确定了其对骨髓来源和脾树突状细胞(DC)的影响。具体来说,我们应用脂多糖(LPS)或β2GPI刺激,并测量CD80和CD86的表达以及细胞因子的分泌。我们还在OT-II小鼠中确定,如果骨髓来源的DC能够刺激抗原特异性T细胞。此外,我们在依赖于β2GPI主动免疫的APS小鼠模型中检测了克拉索利免疫后的治疗潜力.根据肠系膜微血管中荧光素诱导的血栓来评估血管表现,而产科表现是根据妊娠后胎儿丢失的比例进行评估的。我们还测量了抗β2GPI抗体的血液滴度,离体β2GPI刺激后脾细胞增殖反应和细胞因子分泌。最后,我们在这些老鼠身上确定,血液学,Crassolide的肝和肾毒性。Crassolide在LPS刺激后抑制DC成熟和肿瘤坏死因子(TNF)-α的分泌,白细胞介素(IL)-6,IL-12和IL-23,以及下游T细胞活化。Crassolide可以部分改善BALB/c小鼠APS的血管和产科表现。β2GPI刺激后,抗β2GPI抗体的血液滴度和脾细胞增殖均降低。β2GPI刺激后脾Th1和Th17反应也降低。最后,在治疗剂量的克拉索利,我们没有发现其毒性的证据.总之,我们显示了crassolide抑制DC和下游T细胞反应的能力。因此,Crassolide是APS辅助治疗的潜在候选者。
Antiphospholipid syndrome (APS) is an autoimmune disease characterized by the production of β2-glycoprotein I (β2GPI)-dependent autoantibodies, with vascular thrombosis or obstetrical complications. Around 20% of APS patients are refractory to current treatments. Crassolide, a cembranoid diterpene extracted from soft corals, is a potential therapeutic candidate. Here, to examine the anti-inflammatory properties of crassolide, we first determined its effects on bone marrow-derived and splenic dendritic cells (DC). Specifically, we applied lipopolysaccharide (LPS) or β2GPI stimulation and measured the expressions of CD80 and CD86, and secretions of cytokines. We also determined in the OT-II mice, if bone marrow-derived DC was able to stimulate antigen-specific T cells. Moreover, we examined the therapeutic potential of crassolide postimmunization in a murine model of APS that depended on active immunization with β2GPI. The vascular manifestations were evaluated in terms of fluorescein-induced thrombi in mesenteric microvessels, whereas the obstetric manifestations were evaluated based on the proportion of fetal loss after pregnancy. We also measured blood titers of anti-β2GPI antibody, splenic cell proliferative responses and cytokine secretions after β2GPI stimulation ex vivo. Finally, we determined in these mice, hematological, hepatic and renal toxicities of crassolide. Crassolide after LPS stimulation suppressed DC maturation and secretion of tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-12 and IL-23, and downstream T cell activation. Crassolide could partially ameliorate both the vascular and obstetric manifestations of APS in BALB/c mice. Both blood titers of anti-β2GPI antibody and splenic cell proliferation after β2GPI stimulation were reduced. Splenic Th1 and Th17 responses were also lowered after β2GPI stimulation. Finally, within therapeutic doses of crassolide, we found no evidence of its toxicity. In conclusion, we showed the ability of crassolide to suppress DC and downstream T cell responses. Crassolide is therefore a potential candidate for adjunctive therapy in APS.