此前已有研究證實,在蠕蟲感染背景下,有多種細胞和分子機制可促進宿主耐受性,從而防止免疫病理損傷。其中包括M2型巨噬細胞的作用以及神經免疫調節回路。盡管取得了這些進展,但在全球傳播寄生蟲感染之一——旋毛蟲(T. spiralis)感染中,促進耐受性的具體通路仍有待明確。因此,我們首先旨在研究旋毛蟲感染腸道期M2型巨噬細胞的作用。感染后第8天,小腸中M2相關標志物Arg1、Retnla和Chil3的表達升高,提示存在感染誘導的M2型巨噬細胞(圖1A)。為評估巨噬細胞對宿主保護作用的貢獻,我們在感染后經靜脈給予小鼠磷酸鹽緩沖液(PBS:P-005)或載有氯膦酸鹽(CL:C-005)的脂質體,以清除多種髓系細胞群體。結果顯示,CL脂質體處理確實耗竭了Ly6C+CD11b+單核細胞和F4/80+CD11b+巨噬細胞(圖1 B–E)。此外,CL脂質體處理還導致小腸中感染誘導的Arg1、Retnla和Chil3表達喪失(圖1A)。無論PBS還是CL處理,感染動物間iNOS表達均無顯著變化(附錄圖S1A)。關鍵在于,接受CL脂質體處理的小鼠較對照組體重下降更明顯,死亡率也更高(圖1 F和G)。上述數據表明,通過CL脂質體遞送清除吞噬細胞會導致腸道內感染誘導的M2型反應喪失,并與宿主發病率和死亡率升高相關。

M2型巨噬細胞的缺失與旋毛蟲感染后發病率和死亡率的增加相關。經靜脈注射磷酸鹽緩沖液(PBS)或載有氯膦酸鹽(CL)脂質體的小鼠被旋毛蟲感染,并于感染后第8天處死。(A)通過RT-qPCR檢測腸道中M2相關標志物的表達。(B和C)單核細胞(Ly6C+CD11b+)以及(D和E)巨噬細胞(F4/80+CD11b+)在脾臟中的存在與否通過流式細胞術測定。細胞群體從CD45hiCD3?CD19?Ly6G?細胞中設門圈出。流式細胞術圖中的數字代表CD45hi細胞的百分比。(F)在整個感染過程中監測體重變化,(G)記錄死亡率。
論文信息:
論文題目:Monocytes maintain central nervous system homeostasis following helminth-induced inflammation
期刊名稱:PNAS
時間期卷:119 (37) e2201645119
DOI: 10.1073/pnas.2201645119
產品信息:
貨號:CP-010-010
規格:10ml+10ml
品牌:Liposoma
產地:荷蘭
名稱:Clodronate Liposomes&Control Liposomes
辦事處:靶點科技
Clodronate Liposomes氯膦酸鹽脂質體外周血單核巨噬細胞。荷蘭Liposoma巨噬細胞清除劑ClodronateLiposomes見刊于PNAS:單核細胞在蠕蟲誘導的炎癥后維持中樞神經系統穩態。

Liposoma巨噬細胞清除劑Clodronate Liposomes氯膦酸二鈉脂質體清除巨噬細胞的材料和方法:
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.
巨噬細胞清除材料和方法文獻截圖:

請輸入賬號
請輸入密碼
請輸驗證碼
以上信息由企業自行提供,信息內容的真實性、準確性和合法性由相關企業負責,化工儀器網對此不承擔任何保證責任。
溫馨提示:為規避購買風險,建議您在購買產品前務必確認供應商資質及產品質量。