中文摘要：
肿瘤相关巨噬细胞 （TAM） 通过促进肿瘤血管生成在肿瘤生长和转移中起关键作用。用包埋在脂质体中的氯膦酸盐 （Clodronate Liposomes） 治疗有效地耗尽了小鼠 F9 畸胎癌和人 A673 横纹肌肉瘤小鼠肿瘤模型中的这些吞噬细胞，导致肿瘤生长的显着抑制，范围为 75% 至 >92%，具体取决于治疗和时间表。肿瘤抑制伴随着肿瘤组织中血管密度的急剧降低。血管内皮生长因子 （VEGF） 是肿瘤血管生成的主要诱导剂之一，也是巨噬细胞募集所必需的。氯膦盐脂质体和 VEGF 中和抗体的联合治疗观察到效果为优，而游离氯膦酸盐没有显著活性。肿瘤的免疫组织学评估显示 F4/80+ 和 MOMA-1+ 显著耗竭，而 CD11b+ TAM 耗竭不太明显。A673 模型中血管染色 （CD31） 和血管以及 TAM 和肿瘤相关树突状细胞 （TADC） 的定量显示，即使在治疗结束后 9 天，复位率也为 85% 至 >94%。此外，CD11c+ TADC 已被证明在肿瘤释放的生长和分化因子刺激后可能分化为内皮样细胞，氯膦盐脂质体或抗体治疗同样降低了 CD11c+ TADC。这些结果验证了氯膦酸盐脂质体 （Clodronate Liposomes） 疗法与血管生成抑制剂联合使用是一种很有前途的新型癌症治疗策略，用于针对刺激肿瘤生长和播散的造血前体细胞，并作为研究巨噬细胞和树突状细胞在肿瘤发生中的作用的工具。
英文摘要：
Tumour-associated macrophages, TAMs, play a pivotal role in tumour growth and metastasis by promoting tumour angiogenesis. Treatment with clodronate encapsulated in liposomes (clodrolip) efficiently depleted these phagocytic cells in the murine F9 teratocarcinoma and human A673 rhabdomyosarcoma mouse tumour models resulting in significant inhibition of tumour growth ranging from 75 to >92%, depending on therapy and schedule. Tumour inhibition was accompanied by a drastic reduction in blood vessel density in the tumour tissue. Vascular endothelial growth factor (VEGF) is one of the major inducers of tumour angiogenesis and is also required for macrophage recruitment. The strongest effects were observed with the combination therapy of clodrolip and a VEGF-neutralising antibody, whereas free clodronate was not significantly active. Immunohistologic evaluation of the tumours showed significant depletion of F4/80+ and MOMA-1+ and a less pronounced depletion of CD11b+ TAMs. Blood vessel staining (CD31) and quantification of the vessels as well as TAMs and tumour-associated dendritic cells (TADCs) in the A673 model showed reduction rates of 85 to >94%, even 9 days after the end of therapy. In addition, CD11c+ TADCs, which have been shown to potentially differentiate into endothelial-like cells upon stimulation by tumour released growth and differentiation factors, were similarly reduced by clodrolip or antibody treatment. These results validate clodrolip therapy in combination with angiogenesis inhibitors as a promising novel strategy for an indirect cancer therapy aimed at the haematopoietic precursor cells that stimulate tumour growth and dissemination and as a tool to study the role of macrophages and dendritic cells in tumorigenesis.
论文信息：
论文题目： Clodronate-liposome-mediated depletion of tumour-associated macrophages: a new and highly effective antiangiogenic therapy approach
期刊名称：British Journal of Cancer
时间期卷： volume 95, pages272–281 (2006)
在线时间：2006年7月11日
DOI： doi.org/10.1038/sj.bjc.6603240
Liposoma巨噬细胞清除剂氯膦酸盐脂质体Clodronate Liposomes见刊于BJC：
Liposoma巨噬细胞清除剂氯膦酸盐脂质体Clodronate Liposomes的材料和方法：
氯膦酸盐脂质体介导的肿瘤相关巨噬细胞(TAM)耗竭：一种新的高效抗血管生成治疗方法，巨噬细胞清除解决方案
Exponentially growing F9 teratocarcinoma (7 × 106 50 μl−1) or A673 rhabdomyosarcoma cells (6–8 × 106 50 μl−1 mixed 1 : 1, v v−1 with Matrigel, Beckton Dickinson, Basel, Switzerland) were injected subcoutanously (s.c.) on the flanks of mice. Treatment was started 6 h after inoculation of F9 cells (female Sv129 mice) and 24 h after inoculation of A673 cells (female CD-1 nude mice), respectively. The mice (6–8/group) received clodronate dissolved in phosphate buffer (PB, 67 mM, pH 7.4) or clodrolip by intraperitoneal (i.p.) injection as initial dose of 2 mg 20 g−1 mouse body weight, followed by 1 mg for the subsequent doses. The Abs were given at 0.5 mg 20 g−1 mouse body weight in 100 μl PB by intravenous (i.v.) injection into the tail vein. Tumour growth was measured in a blinded fashion with a caliper and volumes were calculated using the equation: V=πab2/6 (a=largest tumour diameter, b=perpendicular diameter). Relative percentual tumour growth was normalised to day one. Mice were killed 8–22 days after onset of treatment and tumours and spleens removed for histology.
6-8周，20g小鼠，首剂量腹腔注射2mg，按Liposoma产品货号CP-005-005，规格是5ml+5ml。浓度是5mg/ml。相当于注射了400ul，后期是1mg剂量维持，也就是注射200ul。
The in vitro cytotoxicity of clodronate was assessed as described before . Briefly, cells were incubated in 96-well plates with liposomes, clodronate and clodrolip (6 h, 37°C, 1 mg clodronate ml−1) and cell viability was determined by addition of WST-1 reagent (Roche Diagnostics, Mannheim, Germany) according to the manufacturer's recommendations.
