Department of Integrative Bioscience and Biotechnology, Institute of Anticancer Medicine Development, Sejong University, 209 Neungdong-ro, Gwangjin-gu, Seoul, 05006, South Korea.
Department of Biomedical Laboratory Science, College of Health and Biomedical Services, Sangji University, Wonju, 26339, South Korea.
Inflamm Res. 2024 Sep;73(9):1511-1527. doi: 10.1007/s00011-024-01915-3. Epub 2024 Jul 19.
Invariant natural killer T (iNKT) cells play protective or pathogenic roles in a variety of immune and inflammatory diseases. However, whether iNKT cells contribute to the progression of acute neuroinflammation remains unclear. Thus, we addressed this question with a mouse model of lipopolysaccharide (LPS)-induced acute neuroinflammation.
For induction of acute neuroinflammation, wild-type (WT) C57BL/6 (B6) mice were injected intraperitoneally (i.p.) with LPS for either three or five consecutive days, and then these mice were analyzed for brain-infiltrating leukocytes or mouse behaviors, respectively. To examine the role of iNKT cell activation in LPS-induced neuroinflammation, mice were injected i.p. with the iNKT cell agonist α-galactosylceramide (α-GalCer) seven days prior to LPS treatment. Immune cells infiltrated into the brain during LPS-induced neuroinflammation were determined by flow cytometry. In addition, LPS-induced clinical behavior symptoms such as depressive-like behavior and memory impairment in mice were evaluated by the open field and Y-maze tests, respectively.
We found that iNKT cell-deficient Jα18 mutant mice display delayed disease progression and decreased leukocyte infiltration into the brain compared with WT mice, indicating that iNKT cells contribute to the pathogenesis of LPS-induced neuroinflammation. Since it has been reported that pre-treatment with α-GalCer, an iNKT cell agonist, can convert iNKT cells towards anti-inflammatory phenotypes, we next explored whether pre-activation of iNKT cells with α-GalCer can regulate LPS-induced neuroinflammation. Strikingly, we found that α-GalCer pre-treatment significantly delays the onset of clinical symptoms, including depression-like behavior and memory impairment, while decreasing brain infiltration of pro-inflammatory natural killer cells and neutrophils, in this model of LPS-induced neuroinflammation. Such anti-inflammatory effects of α-GalCer pre-treatment closely correlated with iNKT cell polarization towards IL4- and IL10-producing phenotypes. Furthermore, α-GalCer pre-treatment restored the expression of suppressive markers on brain regulatory T cells during LPS-induced neuroinflammation.
Our findings provide strong evidence that α-GalCer-induced pre-activation of iNKT cells expands iNKT10 cells, mitigating depressive-like behaviors and brain infiltration of inflammatory immune cells induced by LPS-induced acute neuroinflammation. Thus, we suggest the prophylactic potential of iNKT cells and α-GalCer against acute neuroinflammation.
固有自然杀伤 T(iNKT)细胞在多种免疫和炎症性疾病中发挥保护或致病作用。然而,iNKT 细胞是否有助于急性神经炎症的进展尚不清楚。因此,我们使用脂多糖(LPS)诱导的急性神经炎症小鼠模型来解决这个问题。
为了诱导急性神经炎症,将野生型(C57BL/6,B6)小鼠腹膜内(i.p.)注射 LPS,连续 3 或 5 天,然后分别分析大脑浸润的白细胞或小鼠行为。为了研究 iNKT 细胞激活在 LPS 诱导的神经炎症中的作用,在 LPS 处理前 7 天,将小鼠腹膜内注射 iNKT 细胞激动剂α-半乳糖神经酰胺(α-GalCer)。通过流式细胞术测定 LPS 诱导的神经炎症中浸润到大脑的免疫细胞。此外,通过旷场和 Y 迷宫试验分别评估 LPS 诱导的临床行为症状,如抑郁样行为和记忆障碍。
我们发现,与野生型小鼠相比,iNKT 细胞缺陷型 Jα18 突变小鼠显示出疾病进展延迟和大脑白细胞浸润减少,表明 iNKT 细胞参与了 LPS 诱导的神经炎症的发病机制。由于先前的报道表明,iNKT 细胞激动剂α-GalCer 的预处理可以将 iNKT 细胞转化为抗炎表型,我们接下来探讨了 α-GalCer 预先激活 iNKT 细胞是否可以调节 LPS 诱导的神经炎症。令人惊讶的是,我们发现,α-GalCer 预处理显著延迟了临床症状的发作,包括抑郁样行为和记忆障碍,同时减少了大脑中促炎自然杀伤细胞和中性粒细胞的浸润,在这种 LPS 诱导的神经炎症模型中。α-GalCer 预处理的这种抗炎作用与 iNKT 细胞向产生 IL4 和 IL10 的表型极化密切相关。此外,α-GalCer 预处理恢复了 LPS 诱导的神经炎症期间大脑调节性 T 细胞上抑制性标记物的表达。
我们的研究结果为α-GalCer 诱导的 iNKT 细胞预先激活扩大 iNKT10 细胞,减轻 LPS 诱导的急性神经炎症引起的抑郁样行为和大脑炎症免疫细胞浸润提供了有力证据。因此,我们建议 iNKT 细胞和 α-GalCer 对急性神经炎症具有预防作用。