Real-time monitoring of in vivo acute necrotic cancer cell death induced by near infrared photoimmunotherapy using fluorescence lifetime imaging.
Real-time monitoring of in vivo acute necrotic cancer cell death induced by near infrared photoimmunotherapy using fluorescence lifetime imaging.
复制标题
DOI：
10.1158/0008-5472.can-12-1298
复制
发表时间：
2012-09-15
期刊：
影响因子：
11.2
通讯作者：
Kobayashi H
中科院分区：
文献类型：
--
作者：
Nakajima T;Sano K;Mitsunaga M;Choyke PL;Kobayashi H
最近已经描述了一种新型的基于单克隆抗体（mAb）的高度特异性光疗（光免疫疗法; PIT），其利用与mAb缀合的近红外（NIR）酞菁染料IRDye 700DX（IR 700）。NIR光暴露导致体外立即靶向选择性坏死细胞死亡。检测体内细胞立即死亡更困难，因为肿瘤开始缩小至少需要3天。在本研究中，在PIT之前和之后评估荧光寿命（FLT）以监测NIR介导的mAb-IR 700 PIT的即时细胞毒性效应。抗EGFR帕尼单抗-IR 700用于靶向表达EGFR的A431肿瘤细胞。在体外的细胞团块中和在体内的小鼠皮下异种移植肿瘤中，用各种剂量的NIR光进行PIT。在PIT前和PIT后0、6、24和48 h测量FLT。在体外，在较高剂量的NIR光下的PIT立即导致A431细胞中的FLT缩短。在体内，在阈值NIR剂量为30 J/cm 2或更高后，PIT诱导治疗肿瘤中的FLT立即缩短。相比之下，较低水平的近红外光（10 J/cm 2或更小）不会诱导FLT缩短。在PIT后6小时观察到肿瘤部位周围组织中的FLT延长，可能反映了巨噬细胞的吞噬作用。总之，FLT成像可用于监测mAb-IR 700诱导的PIT的急性细胞毒性作用，甚至在靶向肿瘤中可见形态学变化之前。
A new type of monoclonal antibody (mAb)-based, highly specific phototherapy (photoimmunotherapy; PIT) that utilizes a near infrared (NIR) phthalocyanine dye, IRDye700DX (IR700) conjugated with a mAb, has recently been described. NIR light exposure leads to immediate, target-selective necrotic cell death in vitro. Detecting immediate in vivo cell death is more difficult because it takes at least 3 days for the tumor to begin to shrink in size. In this study, fluorescence lifetime (FLT) was evaluated before and after PIT for monitoring the immediate cytotoxic effects of NIR mediated mAb-IR700 PIT. Anti-EGFR panitumumab-IR700 was used for targeting EGFR-expressing A431 tumor cells. PIT with various doses of NIR light was performed in cell pellets in vitro and in subcutaneously xenografted tumors in mice in vivo. FLT measurements were obtained before and 0, 6, 24 and 48 h after PIT. In vitro, PIT at higher doses of NIR light immediately led to FLT shortening in A431 cells. In vivo PIT induced immediate shortening of FLT in treated tumors after a threshold NIR dose of 30J/cm2 or greater. In contrast, lower levels of NIR light (10J/cm2 or smaller) did not induce shortening of FLT. Prolongation of FLT in tissue surrounding the tumor site was noted 6 hours after PIT, likely reflecting phagocytosis by macrophages. In conclusion, FLT imaging can be used to monitor the acute cytotoxic effects of mAb-IR700-induced PIT even before morphological changes can be seen in the targeted tumors.