单克隆抗体4E5的^131I标记及^131I-4E5对B细胞淋巴瘤细胞的杀伤效应  

An experimental study on labeling monoclonal antibody 4E5 with ^131I and the lethal effects of ×^131I- 4E5 against B cell lymphoma

作　　者：翟士军[1] 邱王华[2] 万卫星[1] 苏成海[3] 陈昌友[2] 杜晓庆[1] 吴娜静[1] 

机构地区：[1]苏州大学附属第四医院、无锡市第四人民医院核医学科,214062 [2]苏州大学医学部免疫学系 [3]苏州大学附属第一医院核医学科

出　　处：《中华核医学与分子影像杂志》2012年第3期218-221,共4页Chinese Journal of Nuclear Medicine and Molecular Imaging

摘　　要：目的研究CD80单克隆抗体4E5的^131I标记及标记产物^131I-4E5对B细胞淋巴瘤细胞的杀伤作用，为B细胞淋巴瘤的放射免疫导向治疗提供实验依据。方法采用Iodogen法对4E5进行^131I标记，以三氯醋酸法测定标记率和放化纯，并对^131I-4E5的免疫活性及稳定性进行分析。以四甲基偶氮唑蓝（MTT）比色法观察^13114E5和4E5对B细胞淋巴瘤Raji细胞的杀伤效应。采用SPSS13．0软件对数据进行单因素方差分析和t检验。结果^131I-4E5的标记率为（78．3±2．4）％，放化纯为（95．7±1．8）％，比活度为0．58MBq／lig，放射性浓度为3．90×10^10Bq／L。^131I-4E5加入血清中放置3d后，放化纯仍〉90％。^131I-4E5与Raji细胞的最大结合率为（36．06±2．63）％；^131I-4E5对Raji细胞的杀伤作用呈剂量依赖性，高剂量组的细胞杀伤效应明显强于低剂量组：放射性浓度为1．48×10^10、7．40×10^9、3．70×10^9、1．85×10^9和9．25×10^8 Bq/L时，细胞抑制率分别为（52．98±5．19）％、（46．29±2．80）％、（41．05±4．83）％、（33．68±3．79）％和（17．89±2．78）％，F=33．882，P〈0．001；4E5组（质量浓度为20．0、10．0、5．0、2．5和1．25mg／L）的细胞抑制率分别为（32．98±3．99）％、（30．88±3．98）％、（27．14±2．05）％、（20．35±4．38）％和（8．42±1．05）％，^131I-4E5组显著高于4E5组（t=5．290、5．489、4．596、3．986和5．515，P均〈0．05）。结论Iodogen法^131I标记4E5标记率和放化纯高，标记物稳定性好；^131I-4E5可与Rajj细胞特异性结合并发挥杀伤效应，为以B细胞淋巴瘤CD80为靶点的放射免疫显像及治疗提供了可能性。Objective To label the monoclonal antibody 4E5 with ^131I and to evaluate the lethal effects of ^131I-4E5 against B cell lymphoma. Methods 4E5 was radiolabeled with ^131I using the Iodogen method at room temperature. The labeling efficiency and radiochemical purity was measured with trichloroacetic acid （TCA） precipitation, and the immune activity and stability of ^131I-4E5 was analyzed. The lethal effects of ^131I-4E5 and 4E5 against Raji ceils were evaluated by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyhe-trazolium bromide （MTF） assay. Analysis of variance and t-test were used for data analysis with SPSS 13.0. Results The labeling rate of ^131 I-4E5 was （ 78.3 ± 2.4 ） % , and the radiochemical purity was （95.7 ± 1.8）%. Its specific activity and radioactive concentration were 0.58 MBq/μg and 3.90 ×10^10±Bq/L, respectively. The radiochemical purity of ^131I-4E5 mixed with serum and PBS was over 90% after three days. The maximum specific binding efficiency of ^131I-4E5 with Raji cells was （36.06 ±2.63）%. ^131I- 4E5 exhibited a dose-dependent cytotoxicity against Raji cells. The lethal effect of the high dose group was significantly stronger than that of the low dose group. When the radioactive concentrations were 1.48 × 10^10 , 7.40 × 10^9 , 3.70 × 10^9 , 1.85× 10^9 and 9.25 × 10^8 Bq/L, the cell inhibition ratios were （52.98 ± 5. 19）%, （46.29±2.80）%, （41.05±4.83）%, （33.68±3.79）% and （17.89±2.78）%, respectively （F = 33. 882, P 〈0.001 ）. In the 4E5 group, when concentrations of 4E5 were 20.0, 10.0, 5.0, 2.5 and 1.25 mg/L, the cell inhibition ratios were （32.98 ± 3.99） %, （30.88 ± 3.98 ） %, （27. 14 ± 2.05 ） % , （ 20.35 ±4.38） % and （ 8.42± 1.05 ） %, respectively. Accordingly, significantly higher growth inhibition rates for Raji cells than 4E5 at all antibody concentrations were tested （t -- 5. 290,5.489, 4.596, 3. 986 and 5.515, all P 〈 0.05）. Conclusions The labeling efficiency and radiochemical purity of 131 I-4E5 using

关 键 词：抗体 单克隆 同位素标记 淋巴瘤 B细胞 碘放射性同位素 放射免疫疗法 

分 类 号：R733.1[医药卫生—肿瘤]