中文摘要：
氯膦酸盐脂质体 （Clo-Lip，Liposoma，CAT：CP-005-005） 已被广泛用于消耗单核吞噬细胞 （MoPh），以研究这些细胞在体内的功能。在这里，我们重新审视了 Clo-Lip 的作用以及 MoPh 缺乏症的遗传模型，揭示了 CloLip 独立于 MoPh 发挥其抗炎作用。值得注意的是，不仅 MoPh 而且多型核中心粒细胞 （PMN） 在体内摄入 Clo-Lip，导致它们的功能停滞。PMN 的过继转移，而不是 MoPh，逆转了 Clo-Lip 治疗的抗炎作用，表明 PMN 的stunning而不是 MoPh 的消耗解释了 Clo-Lip 在体内的抗炎作用。我们的数据强调了对当前关于 MoPh 在炎症中的作用的文献进行关键修订的必要性。
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.
论文信息：
论文题目： Stunning of neutrophils accounts for the anti-inflammatory effects of clodronate liposomes
期刊名称：JEM- J Exp Med
时间期卷： J Exp Med (2023) 220 (6): e20220525.
在线时间：2023年3月28日
DOI： doi.org/10.1084/jem.20220525
Liposoma巨噬细胞清除剂Clodronate Liposomes见刊于JEM：
Liposoma巨噬细胞清除剂Clodronate Liposomes的材料和方法：
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.
