Department of Pediatrics, Faculty of Medicine, Kuwait University, Kuwait City, Kuwait.
Department of Physiology, College of Medicine and Medical Sciences, Arabian Gulf University, Manama, Bahrain.
PLoS One. 2021 May 10;16(5):e0248455. doi: 10.1371/journal.pone.0248455. eCollection 2021.
The Immune System-Released Activating Agent (ISRAA) was discovered as a novel molecule that functions as a mediator between the nervous and immune systems in response to a nervous stimulus following an immune challenge. This research investigated the role of ISRAA) in promoting the ontogeny of the mouse brain astrocytes. Astrocyte cultures were prepared from two-month-old BALB/c mice. Recombinant ISRAA protein was used to stimulate astrocyte cultures. Immunohistochemistry and ELISA were utilized to measure ISRAA and IFN-γ levels, IFN-γR expression and STAT1 nuclear translocation. MTT-assay was used to evaluate cellular survival and proliferation. To assess astrocyte cell lysates and tyrosine-phosphorylated proteins, SDS-PAGE and western blot were used. ISRAA was highly expressed in mouse embryonic astrocytes, depending on cell age. Astrocytes aged seven days (E7) showed increased proliferation and diminished differentiation, while 21-day-old (E21) astrocytes depicted reversed effects. IFN-γ was involved in the ISRAA action as ISRAA induced IFN-γ in both age groups, but only E21 astrocytes expressed IFN-γR. ISRAA stimulation of E21 resulted in tyrosine phosphorylation of numerous cellular proteins and the nuclear translocation of STAT1, a signalling pathway utilized by IFN-γ. The results suggest that ISRAA is involved in mouse brain development through the cytokine network involving IFN-γ.
免疫系统释放的激活剂(ISRAA)是一种新型分子,它作为神经系统和免疫系统之间的中介,在免疫挑战后对神经刺激做出反应。本研究探讨了 ISRAA 在促进小鼠大脑星形胶质细胞发生中的作用。从两个月大的 BALB/c 小鼠中制备星形胶质细胞培养物。使用重组 ISRAA 蛋白刺激星形胶质细胞培养物。免疫组织化学和 ELISA 用于测量 ISRAA 和 IFN-γ 水平、IFN-γR 表达和 STAT1 核易位。MTT 测定用于评估细胞存活和增殖。评估星形胶质细胞细胞裂解物和酪氨酸磷酸化蛋白时,使用 SDS-PAGE 和 Western blot。ISRAA 在小鼠胚胎星形胶质细胞中高度表达,依赖于细胞年龄。七天龄(E7)的星形胶质细胞显示出增殖增加和分化减少,而 21 天大的(E21)星形胶质细胞则呈现相反的效果。IFN-γ 参与 ISRAA 作用,因为 ISRAA 在两个年龄组中诱导 IFN-γ,但只有 E21 星形胶质细胞表达 IFN-γR。ISRAA 刺激 E21 导致许多细胞蛋白的酪氨酸磷酸化和 STAT1 的核易位,这是 IFN-γ 利用的信号通路。结果表明,ISRAA 通过涉及 IFN-γ 的细胞因子网络参与小鼠大脑发育。