此前已有研究证实，在蠕虫感染背景下，有多种细胞和分子机制可促进宿主耐受性，从而防止免疫病理损伤。其中包括M2型巨噬细胞的作用以及神经免疫调节回路。尽管取得了这些进展，但在全球传播寄生虫感染之一——旋毛虫（T. spiralis）感染中，促进耐受性的具体通路仍有待明确。因此，我们首先旨在研究旋毛虫感染肠道期M2型巨噬细胞的作用。感染后第8天，小肠中M2相关标志物Arg1、Retnla和Chil3的表达升高，提示存在感染诱导的M2型巨噬细胞（图1A）。为评估巨噬细胞对宿主保护作用的贡献，我们在感染后经静脉给予小鼠磷酸盐缓冲液（PBS：P-005）或载有氯膦酸盐（CL：C-005）的脂质体，以清除多种髓系细胞群体。结果显示，CL脂质体处理确实耗竭了Ly6C+CD11b+单核细胞和F4/80+CD11b+巨噬细胞（图1 B–E）。此外，CL脂质体处理还导致小肠中感染诱导的Arg1、Retnla和Chil3表达丧失（图1A）。无论PBS还是CL处理，感染动物间iNOS表达均无显著变化（附录图S1A）。关键在于，接受CL脂质体处理的小鼠较对照组体重下降更明显，死亡率也更高（图1 F和G）。上述数据表明，通过CL脂质体递送清除吞噬细胞会导致肠道内感染诱导的M2型反应丧失，并与宿主发病率和死亡率升高相关。
M2型巨噬细胞的缺失与旋毛虫感染后发病率和死亡率的增加相关。经静脉注射磷酸盐缓冲液（PBS）或载有氯膦酸盐（CL）脂质体的小鼠被旋毛虫感染，并于感染后第8天处死。（A）通过RT-qPCR检测肠道中M2相关标志物的表达。（B和C）单核细胞（Ly6C+CD11b+）以及（D和E）巨噬细胞（F4/80+CD11b+）在脾脏中的存在与否通过流式细胞术测定。细胞群体从CD45hiCD3−CD19−Ly6G−细胞中设门圈出。流式细胞术图中的数字代表CD45hi细胞的百分比。（F）在整个感染过程中监测体重变化，（G）记录死亡率。
论文信息：
论文题目：Monocytes maintain central nervous system homeostasis following helminth-induced inflammation
期刊名称：PNAS
时间期卷：119 (37) e2201645119
DOI: 10.1073/pnas.2201645119
产品信息：
货号：CP-010-010
规格：10ml+10ml
品牌：Liposoma
产地：荷兰
名称：Clodronate Liposomes&Control Liposomes
办事处：靶点科技
Clodronate Liposomes氯膦酸盐脂质体外周血单核巨噬细胞。荷兰Liposoma巨噬细胞清除剂ClodronateLiposomes见刊于PNAS：单核细胞在蠕虫诱导的炎症后维持中枢神经系统稳态。
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：
In vivo macrophage depletion
For depletion of CCR2+ monocytes, CCR2-DTR mice and littermate controls were handled as published previously (34). The generalized depletion of macrophages and other phagocytic cells were executed based on the protocol from Liposoma. At necropsy, single-cell suspensions of mesenteric lymph nodes and spleens, and intestinal worm counts were processed as previously described (83). Whole blood was collected in BD Vacutainer Glass Blood Collection Tubes with K3 EDTA, and plasma was isolated for later analysis. Sections of brain, small intestine, kidney, liver, and gastrocnemius muscle were collected in RNAlater buffer for RT-qPCR analysis and formalin for histological analysis (H&E) and immunofluorescent microscopy. ImageJ (v1.52a) software was used to measure cross-sectional areas of muscle fibers from the gastrocnemius (84).
For the adoptive transfer of monocytes to monocyte-depleted animals, monocytes were isolated from infected WT mice bone marrow and spleen 2, 3, 4, 5, and 6 dpi, and then sort-purified. One to one and a half million cells were given to each DTR recipient through retro-orbital vein intravenous injection. Control animals received same volume of sterile PBS.
巨噬细胞清除材料和方法文献截图：
