Institute for Inflammation Research, Center for Rheumatology and Spine Diseases, Copenhagen University Hospital, Rigshospitalet, Copenhagen, Denmark.
Copenhagen Lupus and Vasculitis Clinic, Center for Rheumatology and Spine Diseases, Copenhagen University Hospital, Rigshospitalet, Copenhagen, Denmark.
Clin Exp Immunol. 2021 Apr;204(1):64-77. doi: 10.1111/cei.13569. Epub 2021 Jan 25.
Circulating microvesicles (MVs) from patients with systemic lupus erythematosus (SLE) express the type 1 interferon (IFN)-inducible protein galectin-3 binding protein (G3BP), which may enhance their deposition in the glomerular basement membrane. The release of G3BP-expressing MVs from normal peripheral blood mononuclear cells (PBMCs) is induced by Toll-like receptor 9 (TLR-9) ligands, and these vesicles contain autoantibody-accessible double-stranded DNA (dsDNA). This study compares the release of MVs expressing G3BP and dsDNA from PBMCs derived from SLE patients with or without active lupus nephritis (LN) and from healthy donors, and taps further into the potential dependency on IFN-α for their generation and impacts of TLR-7/TLR-9 co-stimulation. PBMCs from 10 healthy donors and 12 SLE patients, six of whom had active LN at study inclusion, were stimulated in-vitro with recombinant human IFN-α and the TLR-9 agonists oligodeoxynucleotide (ODN)2216 or ODN2395 alone or in combination with the TLR-7 agonist gardiquimod. MVs in the supernatants were subsequently isolated by differential centrifugation and their expression of G3BP and dsDNA was quantified by flow cytometry. Stimulation with ODN2395 significantly increased the release of MVs co-expressing G3BP and dsDNA from PBMCs isolated from healthy donors and SLE patients. The expression of G3BP on individual MVs and the proportion of G3BP and dsDNA double-positive MVs released were increased in active LN patients. Neither co-stimulation with gardiquimod nor with the IFN-α inhibitor IN-1 had any effect on the MV release induced by ODN2395. In conclusion, the TLR-9-mediated inducibility of MVs co-expressing G3BP and dsDNA is increased in SLE patients with active LN.
循环微泡 (MVs) 来自系统性红斑狼疮 (SLE) 患者，表达 I 型干扰素 (IFN)-诱导蛋白 galectin-3 结合蛋白 (G3BP)，这可能增强它们在肾小球基底膜中的沉积。正常外周血单核细胞 (PBMC) 中 G3BP 表达 MV 的释放是由 Toll 样受体 9 (TLR-9) 配体诱导的，这些囊泡包含可与自身抗体结合的双链 DNA (dsDNA)。本研究比较了来自有或无活动性狼疮肾炎 (LN) 的 SLE 患者和健康供体的 PBMC 中表达 G3BP 和 dsDNA 的 MV 的释放，并进一步探讨了其产生对 IFN-α的潜在依赖性以及 TLR-7/TLR-9 共刺激的影响。将来自 10 名健康供体和 12 名 SLE 患者的 PBMC 在体外用重组人 IFN-α和 TLR-9 激动剂寡脱氧核苷酸 (ODN)2216 或 ODN2395 单独或与 TLR-7 激动剂 gardiquimod 刺激。随后通过差速离心分离上清液中的 MV，并通过流式细胞术定量其 G3BP 和 dsDNA 的表达。单独刺激 ODN2395 显著增加了从健康供体和 SLE 患者分离的 PBMC 中表达 G3BP 和 dsDNA 的 MV 的释放。在活动性 LN 患者中，单个 MV 上 G3BP 的表达以及 G3BP 和 dsDNA 双阳性 MV 的释放比例均增加。Gardiquimod 的共刺激或 IFN-α抑制剂 IN-1 对 ODN2395 诱导的 MV 释放均无影响。总之，在活动性 LN 的 SLE 患者中，TLR-9 介导的共表达 G3BP 和 dsDNA 的 MV 诱导性增加。
