Center for Environmental Health Sciences, Department of Comparative Biomedical Sciences, College of Veterinary Medicine, Mississippi State University, MS, United States.
Center for Environmental Health Sciences, Department of Comparative Biomedical Sciences, College of Veterinary Medicine, Mississippi State University, MS, United States.
Int Immunopharmacol. 2024 Mar 10;129:111654. doi: 10.1016/j.intimp.2024.111654. Epub 2024 Feb 9.
Previous studies demonstrated that cannabinoids exhibit immunosuppressive effects in experimental autoimmune encephalomyelitis (EAE), the animal model of multiple sclerosis (MS). To ask questions about treatment timing and investigate mechanisms for immune suppression by the plant-derived cannabinoids, cannabidiol (CBD) and Δ-tetrahydrocannabinol (THC), an in vitro peptide stimulation of naive splenocytes (SPLC) was developed to mimic T cell activation in EAE. The peptide was derived from the myelin oligodendrocyte glycoprotein (MOG) protein, which is one component of the myelin sheath. MOG peptide is typically used with an immune adjuvant to trigger MOG-reactive T cells that attack MOG-containing tissues, causing demyelination and clinical disease in EAE. To develop the in vitro model, naïve SPLC were stimulated with MOG peptide on day 0 and restimulated on day 4. Cytokine analyses revealed that CBD and THC suppressed MOG peptide-stimulated cytokine production. Flow cytometric analysis showed that intracellular cytokines could be detected in CD4+ and CD8+ T cells. To determine if intracellular calcium was altered in the cultures, cells were stimulated for 4 days to assess the state of the cells at the time of MOG peptide restimulation. Both cannabinoid-treated cultures had a smaller population of the calcium-positive population as compared to vehicle-treated cells. These results demonstrate the establishment of an in vitro model that can be used to mimic MOG-reactive T cell stimulation in vivo.
先前的研究表明，大麻素在实验性自身免疫性脑脊髓炎（EAE），即多发性硬化症（MS）的动物模型中具有免疫抑制作用。为了探讨治疗时机问题并研究植物来源的大麻素，即大麻二酚（CBD）和Δ-四氢大麻酚（THC）对免疫的抑制机制，开发了一种体外肽刺激幼稚脾细胞（SPLC）的方法，以模拟 EAE 中 T 细胞的激活。该肽源自髓鞘少突胶质细胞糖蛋白（MOG）蛋白，后者是髓鞘的组成部分之一。MOG 肽通常与免疫佐剂一起使用，以触发攻击含有 MOG 的组织的 MOG 反应性 T 细胞，从而在 EAE 中引起脱髓鞘和临床疾病。为了开发体外模型，在第 0 天用 MOG 肽刺激幼稚 SPLC，并在第 4 天再次刺激。细胞因子分析显示 CBD 和 THC 抑制了 MOG 肽刺激的细胞因子产生。流式细胞术分析显示，CD4+和 CD8+T 细胞中可检测到细胞内细胞因子。为了确定细胞内钙是否在培养物中发生改变，用 MOG 肽对细胞进行了 4 天的刺激，以评估 MOG 肽再次刺激时细胞的状态。与 vehicle 处理的细胞相比，两种大麻素处理的培养物中钙阳性群体的比例较小。这些结果表明，已经建立了一种体外模型，可用于模拟体内 MOG 反应性 T 细胞的刺激。
