Bidkhori Hamid Reza, Farshchian Moein, Kazemi Noughabi Mahboubeh, Hassanzadeh Halimeh, Rafatpanah Houshang
Immunology Research Center, Inflammation and Inflammatory Diseases Division, Mashhad University of Medical Sciences, Mashhad, Iran.
Stem Cells and Regenerative Medicine Department, Academic Center for Education, Culture, and Research (ACECR)-Khorasan Razavi, Mashhad, Iran.
Iran J Basic Med Sci. 2023 Feb;26(2):241-247. doi: 10.22038/IJBMS.2022.67353.14771.
The B18R protein encoded by the Vaccinia virus decoys Type 1 interferons and inhibits the activity of several type I IFN members. In vitro transcription protocols benefit from this molecule's involvement in enhancing cell viability by inhibiting interferon signal transduction. As a result of their immunomodulatory properties and potential to regenerate, mesenchymal stromal cells (MSCs) are increasingly considered an alternative treatment for a wide range of immune disorders. In this study, we investigated the modification of expression of several genes involved in immune-related pathways after preconditioning MSCs with two immune stimuli, including poly(I:C) and LPS.
ASCs were isolated and primed with B18R, and after exposure to poly(I:C) and LPS, the expression of the same sets of genes as in the previous experiment was evaluated. Following total RNA isolation from primed cells and cDNA preparation, real-time quantitative PCR was performed for several immunomodulatory and immune-related genes, including , and .
Pretreatment of MSCs with poly(I:C) and LPS significantly increased the expression of all mentioned genes, while upon the B18R challenge followed by poly(I:C) and LPS treatment, they were down-regulated. Finally, it was observed that the relative expression level of has significantly decreased in MSCs+B18R+poly(I:C) and LPS in comparison with these groups without B18R.
The data indicated that the presence of B18R prevents the overexpression of several immune-related genes, which are overexpressed in the inflammatory environment.
痘苗病毒编码的B18R蛋白可诱骗I型干扰素并抑制几种I型干扰素成员的活性。体外转录方案受益于该分子通过抑制干扰素信号转导来提高细胞活力。由于其免疫调节特性和再生潜力,间充质基质细胞(MSC)越来越被认为是治疗多种免疫疾病的替代方法。在本研究中,我们研究了用两种免疫刺激剂(包括聚肌胞苷酸和脂多糖)对MSC进行预处理后,几种参与免疫相关途径的基因表达的变化。
分离脂肪干细胞并用B18R进行预处理,在暴露于聚肌胞苷酸和脂多糖后,评估与先前实验中相同的一组基因的表达。从预处理细胞中分离总RNA并制备cDNA后,对几种免疫调节和免疫相关基因进行实时定量PCR,包括……
用聚肌胞苷酸和脂多糖对MSC进行预处理可显著增加所有上述基因的表达,而在B18R刺激后再进行聚肌胞苷酸和脂多糖处理时,这些基因表达下调。最后,观察到与没有B18R的组相比,在MSC+B18R+聚肌胞苷酸和脂多糖组中……的相对表达水平显著降低。
数据表明,B18R的存在可防止几种在炎症环境中过表达的免疫相关基因的过表达。