靶向CD30单克隆抗体^(64)Cu-NOTA-CD30的淋巴瘤免疫PET显像研究  

ImmunoPET studies of^(64)Cu-labeled CD30 monoclonal antibody^(64)Cu-NOTA-CD30 in lymphoma models

作　　者：杨旭[1] 李翠翠 刘俊 张茗昱 弓建华[2] 苗庆芳[2] 杨吉刚[1] Yang Xu;Li Cuicui;Liu Jun;Zhang Mingyu;Gong Jianhua;Miao Qingfang;Yang Jigang(Department of Nuclear Medicine,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,China;Laboratory of Oncology,Institute of Medicinal Biotechnology,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京友谊医院核医学科,北京100050 [2]中国医学科学院北京协和医学院医药生物技术研究所肿瘤室,北京100050

出　　处：《中华核医学与分子影像杂志》2023年第3期171-176,共6页Chinese Journal of Nuclear Medicine and Molecular Imaging

基　　金：国家自然科学基金(81771860)。

摘　　要：目的制备靶向CD30单克隆抗体(简称单抗)^(64)Cu-1,4,7-三氮杂环壬烷-1,4,7-三乙酸(NOTA)-CD30,无创性可视化评价淋巴瘤CD30的表达。方法通过Western blot评价5种淋巴瘤细胞株(Karpas299、Raji、Daudi、Ramos和U266)中CD30的表达水平。选择高和低表达CD30的细胞株行流式细胞术评估抗CD30单抗特异性结合能力。取NSG小鼠13只构建CD30阳性和阴性皮下荷瘤鼠模型。标记获得^(64)Cu-NOTA-CD30,以^(64)Cu-NOTA-免疫球蛋白(Ig)G为对照探针。经尾静脉注射2种探针后2、24和48 h行microPET显像及生物分布分析。采用重复测量方差分析及Bonferroni法进行数据比较。结果Karpas299细胞呈CD30高表达,Raji细胞呈CD30低表达。流式细胞术示抗CD30单抗与Karpas299细胞特异性结合。^(64)Cu-NOTA-CD30与^(64)Cu-NOTA-IgG的放化纯均>95%。在microPET显像中,Karpas299肿瘤^(64)Cu-NOTA-CD30摄取随时间延长逐渐升高,2、24和48 h分别为(11.46±0.58)、(17.60±1.16)与(19.46±0.99)每克组织百分注射剂量率(%ID/g);48 h时与本底对比度良好,肿瘤与心(血液)比值为2.20±0.22。48 h时,^(64)Cu-NOTA-CD30在Karpas299肿瘤的摄取高于其在Raji肿瘤[(6.10±1.03)%ID/g]及^(64)Cu-NOTA-IgG在Karpas299肿瘤的摄取[(5.12±0.89)%ID/g],差异均有统计学意义(F=290.99,t值:19.65和22.25,均P<0.001)。^(64)Cu-NOTA-CD30与^(64)Cu-NOTA-IgG在各组心、肝的摄取随时间延长逐渐降低。48 h体外生物分布结果与活体microPET显像基本一致。结论^(64)Cu-NOTA-CD30能在活体水平无创性可视化评价淋巴瘤CD30的表达及分布情况,有望应用于靶向CD30免疫治疗的受益群体筛选及疗效评价。Objective To develop the anti-CD30 monoclonal antibody^(64)Cu-1,4,7-trizacyclononane-1,4,7-triacetic acid(NOTA)-CD30 and visualize CD30 expression in lymphoma non-invasively.Methods The CD30 expression levels of 5 cell lines(Karpas299,Raji,Daudi,Ramos,and U266)were assessed by Western blot.Cell lines with high and low CD30 expression were selected for flow cytometry to evaluate the specific binding affinity of anti-CD30 monoclonal antibody.Thirteen NSG mice were used to established CD30 positive and negative subcutaneous xenograft models.^(64)Cu-NOTA-CD30 was obtained and^(64)Cu-NOTA-immunoglobulin(Ig)G was used as the control.ImmunoPET imaging was performed 2,24,and 48 h after the injection of^(64)Cu-NOTA-CD30 or^(64)Cu-NOTA-IgG.Finally,the biodistribution studies were conducted.Repeated-measures analysis of variance and Bonferroni test were conducted for comparison.Results Karpas299 showed the highest CD30 expression,while Raji showed the lowest.Flow cytometry showed specific binding affinity of the anti-CD30 monoclonal antibody to the Karpas299 cell line.The radiochemical purities of the probes were both higher than 95%.In microPET,the^(64)Cu-NOTA-CD30 uptake of Karpas299 xenograft tumors increased over time,with(11.46±0.58),(17.60±1.16)and(19.46±0.99)percentage activity of injection dose per gram of tissue(%ID/g)at 2,24 and 48 h respectively.The contrast to normal tissue was good at 48 h,with the tumor/heart(blood)ratio of 2.20±0.22.The uptake of^(64)Cu-NOTA-CD30 in Karpas299 tumor at 48 h after injection was significantly higher than that in Raji tumor((6.10±1.03)%ID/g)and^(64)Cu-NOTA-IgG in Karpas299 tumor((5.12±0.89)%ID/g;F=290.99,t values:19.65 and 22.25,all P<0.001).The uptake of^(64)Cu-NOTA-CD30 and the control probe in the heart and liver decreased over time in all groups.Ex vivo biodistribution at 48 h was mainly consistent with the results of microPET in vivo.Conclusions^(64)Cu-NOTA-CD30 is able to visualize the expression level and distribution of CD30 non-invasively.It is promising to be appl

关 键 词：淋巴瘤 抗体 单克隆 抗原 CD30 同位素标记 铜放射性同位素 正电子发射断层显像术 肿瘤细胞 培养的 小鼠 

分 类 号：R733.1[医药卫生—肿瘤]