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使用基于HIV的慢病毒载体进行基因工程改造的脂肪间充质干细胞作为自身免疫性疾病的基因疗法

Genetically Engineered Adipose Mesenchymal Stem Cells Using HIV-Based Lentiviral Vectors as Gene Therapy for Autoimmune Diseases.

作者信息

Rostami Masoumeh, Haidari Kamran, Shahbazi Majid

机构信息

1 Department of Molecular Medicine, School of Advanced Technologies in Medicine, Golestan University of Medical Sciences , Gorgan, Iran .

2 Department of Anatomy, Faculty of Medical Sciences, Golestan University of Medical Sciences , Gorgan, Iran .

出版信息

Cell Reprogram. 2018 Dec;20(6):337-346. doi: 10.1089/cell.2018.0006. Epub 2018 Oct 9.

DOI:10.1089/cell.2018.0006
PMID:30307322
Abstract

The immunomodulatory and self-renewable features of human adipose-derived mesenchymal stem cells (hAD-MSCs) mark their importance in regenerative medicine. Interleukin (IL)-23 as a proinflammatory cytokine suppresses T regulatory cells and promotes the response of T helper 17 and T helper 1 cells. This pathway initiates inflammation and immunosuppression in several autoimmune diseases. The current study aimed at producing recombinant IL-23 decoy receptor (RIL-23R) using hAD-MSCs as a good candidate for cell-based gene therapy purposes to reduce inflammation in autoimmune diseases. hAD-MSCs was isolated from lipoaspirate and then characterized by differentiation. RIL-23R was designed and cloned into a pCDH813A-1 lentiviral vector. The transduction of hAD-MSCs was performed at multiplicity of infection = 50 with pCDH-EFI α-RIL-23R-PGK copGFP. Expressions of RIL-23R and octamer-binding transcription factor 4 (OCT-4) were determined by real-time polymerase chain reaction. Self-renewing properties were assayed with OCT-4. Bioactivity of the designed RIL-23R was evaluated by IL-17 and IL-10 expression of mouse splenocytes. The results showed that the transducted hAD-MSCs/RIL-23R, expressing IL-23 decoy receptor, can provide a useful approach for a basic research on cell-based gene therapy for autoimmune disorders.

摘要

人脂肪来源间充质干细胞(hAD-MSCs)的免疫调节和自我更新特性标志着它们在再生医学中的重要性。白细胞介素(IL)-23作为一种促炎细胞因子,可抑制调节性T细胞并促进辅助性T细胞17和辅助性T细胞1的反应。该途径在几种自身免疫性疾病中引发炎症和免疫抑制。当前研究旨在利用hAD-MSCs作为基于细胞的基因治疗的良好候选者来生产重组IL-23诱饵受体(RIL-23R),以减轻自身免疫性疾病中的炎症。从抽脂物中分离出hAD-MSCs,然后通过分化进行表征。设计RIL-23R并将其克隆到pCDH813A-1慢病毒载体中。用pCDH-EFIα-RIL-23R-PGK copGFP以感染复数=50对hAD-MSCs进行转导。通过实时聚合酶链反应测定RIL-23R和八聚体结合转录因子4(OCT-4)的表达。用OCT-4检测自我更新特性。通过小鼠脾细胞的IL-17和IL-10表达评估设计的RIL-23R的生物活性。结果表明,转导的hAD-MSCs/RIL-23R表达IL-23诱饵受体,可为自身免疫性疾病基于细胞的基因治疗的基础研究提供有用的方法。

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