甘露聚糖诱导的银屑病
携带Ncf1基因突变的小鼠经鼻内给予20 mg甘露聚糖,以诱导银屑病和银屑病关节炎(PsA)(24)。采用与前述相同的评分系统,每日对关节炎严重程度进行评分。银屑病样皮肤表现的严重程度通过一套15分制系统进行监测,其中每只耳朵或每个爪子按1至3分的等级进行评估(1分,轻微脱皮;2分,中度脱皮;3分,严重脱皮并伴有部分脱毛)(24)。
对于体内巨噬细胞清除实验,在诱导关节炎前2天,经鼻内给予小鼠50 μl氯膦酸二钠脂质体或对照PBS脂质体(荷兰Liposoma BV公司)。通过流式细胞术分析评估清除效率。
对于体内Ym1蛋白补充实验,在诱导关节炎的第0天经鼻内给予小鼠甘露聚糖,并在诱导后的第0、2和4天经鼻内给予重组Ym1蛋白(每只小鼠1 μg;北京义翘神州生物技术有限公司)。
小鼠肺部炎症模型
对于甘露聚糖诱导的肺部炎症模型,BR.Ncf1和BR.Ncf1.Ym1Δ小鼠经鼻内给予甘露聚糖(每只小鼠0.5 mg)。分别于第0、2和5天处死小鼠。计算肺重与体重的比值(肺指数)。制备肺组织进行苏木精-伊红(H&E)染色。通过流式细胞术分析支气管肺泡灌洗液(BALF)中的炎症细胞。
对于过敏性气道肺部炎症(AIPI)模型的诱导,B10.RIII和BR.Ym1Δ小鼠在第0天和第7天通过腹腔注射0.2 ml乳化液进行致敏,该乳化液由10 μg(溶于100 μl)卵清蛋白(OVA)(美国Sigma-Aldrich公司)和100 μl Imject Alum佐剂(美国Pierce,Thermo Fisher Scientific公司)混合而成。在第14天至第20天期间,小鼠每天接受1% OVA气溶胶激发30分钟。对照组小鼠接受假性致敏,并暴露于相同体积的PBS。在第21天处死小鼠。制备肺组织用于组织学分析和细胞因子表达检测。通过流式细胞术分析BALF中的炎症细胞。收集血清,用于检测Ym1和OVA特异性IgG1及IgE的浓度。
肺泡巨噬细胞清除效果检测

论文信息:
论文题目:Natural polymorphism of Ym1 regulates pneumonitis through alternative activation of macrophages
期刊名称:Science Advances
时间期卷:Vol 6, Issue43(2020)
在线时间:2020年10月21日
DOI: 10.1126/sciadv.aba9337
产品信息:
货号:CP-005-005
规格:5ml+5ml
品牌:Liposoma
产地:荷兰
名称:Clodronate Liposomes&Control Liposomes
办事处:靶点科技
Clodronate Liposomes氯膦酸盐脂质体清除肺炎模型小鼠体内肺泡巨噬细胞。荷兰Liposoma巨噬细胞清除剂ClodronateLiposomes见刊于Science Advances:Ym1的自然多态性通过巨噬细胞的替代性激活来调节肺炎。

Liposoma巨噬细胞清除剂Clodronate Liposomes氯膦酸二钠脂质体清除巨噬细胞的材料和方法:ClodronateLiposoems氯膦酸盐脂质体清除甘露聚糖诱导的肺部炎症模型巨噬细胞模型
In vivo macrophage depletion
Mice with Ncf1 mutation were administrated intranasally with 20 mg of mannan to induce psoriasis and PsA (24) and were scored daily for arthritis severity using the same scoring system as mentioned above. The severity of the psoriasis-like skin manifestations was monitored on a 15-score system, in which ears or each paw was evaluated by a scale ranging from 1 to 3 (1, weak skin peeling; 2, moderate skin peeling; and 3, heavy skin peeling with some hair loss) (24). For the in vivo macrophage depletion experiment, 50 μl of clodronate-liposome or control PBS-liposome (Liposoma BV, The Netherlands) was administrated intranasally to mice 2 days before arthritis induction. The depletion efficiency was assessed by flow cytometry analysis. For the in vivo Ym1 protein supplement experiment, mice were intranasally administrated with mannan to induce arthritis at day 0, and recombinant Ym1 protein (1 μg per mouse; Sino Biological Inc.) was treated intranasally at days 0, 2, and 4 after arthritis induction.
巨噬细胞清除材料和方法文献截图:ClodronateLiposoems氯膦酸盐脂质体清除甘露聚糖诱导的肺部炎症模型巨噬细胞模型