Tan B, Tang H, Xu Y, Chen M J, Wang M Z, Qian J M
Department of Gastroenterology, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Science, Beijing 100730, China.
Department of Internal Medicine, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Science, Beijing 100730, China.
Zhonghua Yi Xue Za Zhi. 2020 Nov 17;100(42):3332-3337. doi: 10.3760/cma.j.cn112137-20200520-01598.
Modeling the immune-related adverse events (irAE) colitis in mice, and explore the protective effect and related mechanism of (LGG) on irAE colitis. C57BL/6 mice were divided into dextran sodium sulfate (DSS) group (=3), DSS+anti-programmed death receptor 1 (PD-1) group (=4), DSS+anti-PD-1+anti-cytotoxic T lymphocyte associated protein 4 (CTLA-4) Group (=4), DSS+anti-PD-1+anti-CTLA-4+LGG group (=4), all were given corresponding drugs and probiotics intervention. The severity of colitis were assessed by weight loss, disease activity index (DAI), colon length, colon histopathological score. The inflammatory cytokines and T cell immunity of CD4, CD8, FoxP3regulatory T cells (Treg), were detected by real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and immunohistochemical staining respectively. Compared to DSS group, the Day 9 weight [(87.40±1.79)% vs (94.57±0.53)%, <0.05], colon length [(5.33±0.27)cm vs (6.63±0.12)cm, <0.05] were lower, and DAI score(2.66±0.24 vs 0.89±0.48), colon histopathological score (12.50±1.04 vs 5.67±0.33), tumor necrosis factor-α (TNF-α) (6.73±1.68 vs 0.91±0.40) (<0.05), as well CD8T cells (156.80±8.84 vs 89.00±6.66) and FoxP3Treg cells (103.80±2.66 vs 48.33±3.18) (<0.05) were higher in DSS+anti-PD-1+anti-CTLA-4 group. Compared to DSS+anti-PD-1+anti-CTLA-4 group, the DAI score(1.83±0.17 vs 2.66±0.24), colonic histopathology score (8.75±0.63 vs 12.50±1.04), TNF-α level (1.32±0.18 vs 6.73±1.68) (<0.05) were lower; and CD8T cells(97.75±3.75 vs 156.80±8.84, <0.01) level was lower with higher FoxP3Treg cells (126.00±8.33 vs 103.80±2.66, =0.046) in DSS+anti-PD-1+anti-CTLA-4+LGG group. DSS combined with anti-PD-1 and anti-CTLA-4 can successfully modeling the irAE colitis in mice, LGG can reduce irAE colitis severity by regulating Treg cells.
建立小鼠免疫相关不良事件（irAE）结肠炎模型，探讨鼠李糖乳杆菌（LGG）对irAE结肠炎的保护作用及相关机制。将C57BL/6小鼠分为葡聚糖硫酸钠（DSS）组（n = 3）、DSS+抗程序性死亡受体1（PD-1）组（n = 4）、DSS+抗PD-1+抗细胞毒性T淋巴细胞相关蛋白4（CTLA-4）组（n = 4）、DSS+抗PD-1+抗CTLA-4+LGG组（n = 4），均给予相应药物及益生菌干预。通过体重减轻、疾病活动指数（DAI）、结肠长度、结肠组织病理学评分评估结肠炎的严重程度。分别采用实时定量逆转录-聚合酶链反应（qRT-PCR）和免疫组织化学染色检测炎性细胞因子以及CD4、CD8、FoxP3调节性T细胞（Treg）的T细胞免疫情况。与DSS组相比，DSS+抗PD-1+抗CTLA-4组第9天体重[（87.40±1.79）% vs （94.57±0.53）%，P<0.05]、结肠长度[（5.33±0.27）cm vs （6.63±0.12）cm，P<0.05]较低，而DAI评分（2.66±0.24 vs 0.89±0.48）、结肠组织病理学评分（12.50±1.04 vs 5.67±0.33）、肿瘤坏死因子-α（TNF-α）（6.73±1.68 vs 0.91±0.40）（P<0.05）以及CD8 T细胞（156.80±8.84 vs 89.00±6.66）和FoxP3 Treg细胞（103.80±2.66 vs 48.33±3.18）（P<0.05）较高。与DSS+抗PD-1+抗CTLA-4组相比，DSS+抗PD-1+抗CTLA-4+LGG组的DAI评分（1.83±0.17 vs 2.66±0.24）、结肠组织病理学评分（8.75±0.63 vs 12.50±1.04）、TNF-α水平（1.32±0.18 vs 6.73±1.68）（P<0.05）较低；CD8 T细胞水平（97.75±3.75 vs 156.80±8.84，P<0.01）较低，而FoxP3 Treg细胞较高（126.00±8.33 vs 103.80±2.66，P = 0.046）。DSS联合抗PD-1和抗CTLA-4可成功建立小鼠irAE结肠炎模型，LGG可通过调节Treg细胞减轻irAE结肠炎的严重程度。
