Non-CpG-containing antisense 2′-methoxyethyl oligonucleotides activate a proinflammatory response independent of toll-like receptor 9 or myeloid differentiation factor 88
Non-CpG-containing antisense 2′-methoxyethyl oligonucleotides activate a proinflammatory response independent of toll-like receptor 9 or myeloid differentiation factor 88
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DOI：
10.1124/jpet.105.084004
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发表时间：
2005-09-01
影响因子：
3.5
通讯作者：
Henry, SP
中科院分区：
文献类型：
--
作者：
Senn, JJ;Burel, S;Henry, SP
具有“CpG”基序的寡核苷酸通过激活 Toll 样受体 9 (TLR9) 引发促炎反应，目前正在研究利用这些特性作为佐剂和癌症治疗。然而，用于反义应用 (ASO) 的寡核苷酸旨在通过避免 CpG 基序和使用化学修饰 [即 2'-甲氧基乙基 (MOE) 糖和 5-甲基胞嘧啶残基] 来最大限度地减少促炎反应。尽管如此，修饰的 ASO 在高剂量下能够引发促炎反应，尽管与 CpG 寡核苷酸相比轻微。为了确定这种现象是否是 TLR 介导的，野生型、TLR9 敲除和骨髓分化因子 88 (MyD88) 敲除小鼠用硫代磷酸酯修饰的寡脱氧核糖核苷酸 CpG 最佳寡核苷酸 (ISIS 12449) 和代表性非 CpG 2'-MOE 寡核苷酸 (ISIS 116847) 进行处理。相对于 ISIS 12449，非 CpG 寡核苷酸的促炎效力较低，在野生型动物中需要高 10 倍的剂量才能引发促炎反应。此外，低剂量 ISIS 12449 的炎症反应依赖于 TLR9 和 MyD88，而非 CpG 寡核苷酸保留了在基因敲除动物中激活促炎症反应的能力。用非CpG寡核苷酸处理的动物表现出脾脏重量增加、细胞因子水平升高、肝脏中免疫细胞浸润增加以及单核细胞/巨噬细胞类型细胞典型的细胞表面标志物的mRNA水平增加。用非 CpG 寡核苷酸处理的野生型和基因敲除动物的骨髓来源细胞的反应与 MIP-2 的产生和细胞外信号调节的 kianse1/2 的激活类似。这些数据暗示非 CpG 2'-MOE 寡核苷酸的 TLR 独立激活机制。
Oligonucleotides with a "CpG" motif trigger a proinflammatory response through activation of Toll-like receptor 9 (TLR9) and are being studied to exploit these properties for use as adjuvants and cancer therapies. However, oligonucleotides intended for antisense applications (ASOs) are designed to minimize proinflammatory responses by avoiding CpG motifs and by using chemical modifications [i.e., 2'-methoxyethyl (MOE) sugars and 5-methyl cytosine residues]. Nonetheless, modified ASOs are capable of eliciting a proinflammatory response at high doses, albeit mild compared with CpG oligos. To determine whether this phenomena is TLR-mediated, wild-type, TLR9 knockout, and myeloid differentiation factor 88 (MyD88) knockout mice were treated with a phosphorothioate-modified oligodeoxyribonucleotide CpG optimal oligo ( ISIS 12449), and a representative non-CpG 2'-MOE oligonucleotide (ISIS 116847). The non-CpG oligonucleotide had a lower proinflammatory potency relative to ISIS 12449, requiring a > 10-fold higher dose in wild-type animals to trigger a proinflammatory response. Furthermore, the inflammatory response to ISIS 12449 at low doses was TLR9 and MyD88-dependent, whereas non-CpG oligonucleotides retained the ability to activate a proinflammatory response in the knockout animals. Animals treated with the non-CpG oligonucleotide exhibited an increased spleen weight, elevated cytokine levels, increased immune cell infiltrates in liver, and an increased level of mRNA for cell surface markers typical of monocyte/macrophage type cells. Bone marrow-derived cells from wild-type and knockout animals treated with non-CpG oligonucleotide responded similarly with the production of MIP-2 and the activation of extracellular signal-regulated kianse1/2. These data implicate a TLR-independent mechanism of activation for non-CpG 2'-MOE oligonucleotides.