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JEM中性粒細(xì)胞被巨噬細(xì)胞清除劑清除單核巨噬細(xì)胞的干擾

時間:2024-11-2閱讀:677

中文摘要:

氯膦酸鹽脂質(zhì)體 (Clo-Lip,Liposoma,CAT:CP-005-005) 已被廣泛用于消耗單核吞噬細(xì)胞 (MoPh),以研究這些細(xì)胞在體內(nèi)的功能。在這里,我們重新審視了 Clo-Lip 的作用以及 MoPh 缺乏癥的遺傳模型,揭示了 CloLip 獨立于 MoPh 發(fā)揮其抗炎作用。值得注意的是,不僅 MoPh 而且多型核中心粒細(xì)胞 (PMN) 在體內(nèi)攝入 Clo-Lip,導(dǎo)致它們的功能停滯。PMN 的過繼轉(zhuǎn)移,而不是 MoPh,逆轉(zhuǎn)了 Clo-Lip 治療的抗炎作用,表明 PMN 的stunning而不是 MoPh 的消耗解釋了 Clo-Lip 在體內(nèi)的抗炎作用。我們的數(shù)據(jù)強(qiáng)調(diào)了對當(dāng)前關(guān)于 MoPh 在炎癥中的作用的文獻(xiàn)進(jìn)行關(guān)鍵修訂的必要性。

英文摘要:

Clodronate liposomes (Clo-Lip, Liposoma, CAT:CP-005-005) have been widely used to deplete mononuclear phagocytes (MoPh) to study the function ofthese cells in vivo. Here, we revisited the effects of Clo-Lip together with genetic models of MoPh deficiency, revealing that CloLip exert their anti-inflammatory effects independent of MoPh. Notably, not only MoPh but also polymorphonuclearneutrophils (PMN) ingested Clo-Lip in vivo, which resulted in their functional arrest. Adoptive transfer of PMN, but not ofMoPh, reversed the anti-inflammatory effects of Clo-Lip treatment, indicating that stunning of PMN rather than depletion ofMoPh accounts for the anti-inflammatory effects of Clo-Lip in vivo. Our data highlight the need for a critical revision of thecurrent literature on the role of MoPh in inflammation.


論文信息:

論文題目: Stunning of neutrophils accounts for the anti-inflammatory effects of clodronate liposomes

期刊名稱:JEM- J Exp Med

時間期卷: J Exp Med (2023) 220 (6): e20220525.

在線時間:2023年3月28日

DOI: doi.org/10.1084/jem.20220525


Liposoma巨噬細(xì)胞清除劑Clodronate Liposomes見刊于JEM:

JEM中性粒細(xì)胞被巨噬細(xì)胞清除劑清除單核巨噬細(xì)胞的干擾


Liposoma巨噬細(xì)胞清除劑Clodronate Liposomes的材料和方法

JEM中性粒細(xì)胞被巨噬細(xì)胞清除劑清除單核巨噬細(xì)胞的干擾

Liposomes

Clodronate liposomes (SKU: C-005) and PBS liposomes(SKU: P-005) were commercially available and purchasedfrom Liposoma BV . The concentration of the clodronatein the suspension was 5 mg/ml. Liposome suspensions were injected directly without any further dilutions. For labeling ofclodronate liposomes with Vybrant DiD (Vybrant DiD Cell-Labeling Solution, Invitrogen, V22887), 10 μl of the Vybrant dyesolution was added to 1 ml clodronate liposome suspension andmixed by gentle shaking. After incubation at 37°C for 20 min(slowly shaking), labeled liposome suspension was centrifuged at1,500 rpm for 5 min, producing a loose phase of liposomes and anupper aqueous phase. The aqueous phase was removed, and anequivalent volume of PBS was added to the liposomes and mixedwell by gentle pipetting. After an additional centrifugation step(1,500 rpm, 5 min) and removal of the upper aqueous phase, anequivalent volume of PBS was added to liposomes to get theoriginal volume.

Cellular depletion approaches

For DT-mediated cell depletion in Cx3cr1cre;iDTR mice, DT(500 ng/mouse) was either injected twice within 24 h starting6 d before arthritis induction for the predepletion or daily(500 ng DT/mouse on the first day, and 100 ng DT/mouse onfollowing days) starting 1 d before arthritis induction for continuous depletion.Clo-Lip (200 μl) were i.v. injected. For imaging and flowcytometry, liposomes were labeled with Vybrant DiD usingVybrant Cell-Labeling solutions. 10 μl of Vybrant dye per mlliposomes was incubated for 20 min at 37°C slowly shakingfollowed by two washing steps with PBS and reconstitution tothe original volume. Arthritis induction, intravital microscopy(IVM), and blood collection for ex vivo stimulation of neutrophils were conducted 24 h after liposome injection.Antibody-mediated depletion was performed by intraperitoneal injection of 500 µg InVivoMab anti-mouse Ly6G antibody(1A8, BioXCell) every 2 d. Isotype injection was used as a control.



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