在人类志愿者外周血单个核细胞中引起物种特异性炎症反应

StephenW

在人类志愿者外周血单个核细胞中引起物种特异性炎症反应的反义寡核苷酸的早期鉴定和避免

塞巴斯蒂安·A·布雷尔
, 托德·马赫默
, 布伦达·F·贝克
, T.杰西·郭
, 苏珊娜·帕兹
, 胡萨姆·尤尼斯
和斯科特·P·亨利
在线发布：2022 年 8 月 17 日 https://doi.org/10.1089/nat.2022.0033

    部分
    查看文章

    权限和引文
    分享

抽象的

开发了一种基于人外周血单核细胞 (PBMC) 的测定法，以鉴定具有激活超出可接受水平的细胞先天免疫反应的潜力的反义寡核苷酸 (ASO)。当 ISIS 353512 靶向人类 C 反应蛋白 (CRP) 的信使核糖核酸在 I 期临床试验中进行测试时，该检测的开发就开始了，其中健康的人类志愿者意外地经历了白细胞介素 6 (IL-6) 的增加和 CRP。在啮齿动物和非人类灵长类动物安全性研究中，没有观察到过度促炎作用的证据，没有预料到这种水平的免疫刺激。 ISIS 353512 诱导的 IL-6 增加是由 B 细胞的激活引起的。 IL-6 诱导受到 PBMC 的氯喹预处理的抑制，并且 ASO 的性质表明该反应是由 Toll 样受体 (TLR) 介导的，很可能是 TLR9。在评估 PBMC 供体间的变异性时，确定了对 ISIS 353512 的两类人类 PBMC 响应者（鉴别者和非鉴别者）。在没有 ASO 处理的情况下，鉴别器供体 PBMC 在培养 24 小时后显示出低水平的 IL-6。使用鉴别器供体的 PBMC 测定被证明是可重复的，通过与以前在临床试验中被证明是安全或炎症的已知基准 ASO 对照进行比较，可以可靠地评估 ASO 的免疫潜力。临床试验注册号：NCT00048321 NCT00330330 NCT00519727。

StephenW

Early-Stage Identification and Avoidance of Antisense Oligonucleotides Causing Species-Specific Inflammatory Responses in Human Volunteer Peripheral Blood Mononuclear Cells

Sebastien A. Burel
, Todd Machemer
, Brenda F. Baker
, T. Jesse Kwoh
, Suzanne Paz
, Husam Younis
, and Scott P. Henry
Published Online:17 Aug 2022https://doi.org/10.1089/nat.2022.0033

    Sections
    View article

    Permissions & Citations
    Share

Abstract

A human peripheral blood mononuclear cell (PBMC)-based assay was developed to identify antisense oligonucleotide (ASO) with the potential to activate a cellular innate immune response outside of an acceptable level. The development of this assay was initiated when ISIS 353512 targeting the messenger ribonucleic acid for human C-reactive protein (CRP) was tested in a phase I clinical trial, in which healthy human volunteers unexpectedly experienced increases in interleukin-6 (IL-6) and CRP. This level of immune stimulation was not anticipated following rodent and nonhuman primate safety studies in which no evidence of exaggerated proinflammatory effects were observed. The IL-6 increase induced by ISIS 353512 was caused by activation of B cells. The IL-6 induction was inhibited by chloroquine pretreatment of PBMCs and the nature of ASOs suggested that the response is mediated by a Toll-like receptor (TLR), in all likelihood TLR9. While assessing the inter PBMC donor variability, two classes of human PBMC responders to ISIS 353512 were identified (discriminator and nondiscriminators). The discriminator donor PBMCs were shown to produce low level of IL-6 after 24 h in culture, in the absence of ASO treatment. The PBMC assay using discriminator donors was shown to be reproducible, allowing to assess reliably the immune potential of ASOs by comparison to known benchmark ASO controls that were previously shown to be either safe or inflammatory in clinical trials. Clinical Trial registration numbers: NCT00048321 NCT00330330 NCT00519727.

StephenW

https://www.liebertpub.com/doi/1 ... +November+9%2C+2022