Internalisation enhances photo-induced cytotoxicity of monoclonal antibody-phthalocyanine conjugates
Internalisation enhances photo-induced cytotoxicity of monoclonal antibody-phthalocyanine conjugates
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DOI：
10.1054/bjoc.2001.2170
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发表时间：
2001-11-30
影响因子：
8.8
通讯作者：
Pèlegrin, A
中科院分区：
文献类型：
--
作者：
Carcenac, M;Dorvillius, M;Pèlegrin, A
癌症免疫光疗将单克隆抗体 (MAb) 对过度表达的肿瘤标志物的特异性与缀合染料的光毒性特性相结合。分析染料内化对光诱导细胞毒性的潜在作用。我们比较了两种靶抗原，不内化的癌胚抗原 (CEA) 和内化的 ErbB2。用 CEA cDNA 转染表达高水平 ErbB2 的人卵巢癌 SKOv3 细胞。使用 FACS 分析，所得细胞系 SKOv3-CEA-1B9 表现出两种靶抗原的表达水平相当。四磺酞菁铝 (AlPcS4) 通过五碳磺酰胺间隔链 (A(1)) 以每摩尔 MAb 6 至 9 摩尔 AlPcS4 的摩尔比与抗 CEA MAb 35A7、抗 ErbB2 MAb FSP77 和非特异性 MAb PX 共价偶联。 35A7-(AlPcS(4)A(1))(8) 缀合物在光照后以 2.50 mug/ml（基于 AlPcS(4)A(1) 含量）孵育 20 小时后，对 SKOv3-CEA-1B9 细胞系产生 68% 的生长抑制。然而，在相同的孵育时间和暴露于相同的光剂量后，FSP77-(AlPcS(4)A(1))(6)缀合物对低至0.04μg/ml的AlPCS(4)A(1)浓度产生51％的生长抑制。在 1.25 mug/ml AlPcS(4)A(1) 浓度下，FSP77-(AlPcS(4)A(1))(6) 缀合物在短至 1 小时的孵育时间后产生 67% 的生长抑制，在 8 小时的孵育时间后达到 96% 的抑制。使用表达两种不同靶抗原的独特细胞系。我们证明了内化相对于非内化 MAb 染料缀合物在光毒性功效方面的明显优势。缀合物光动力学特性的体内评估正在进行中。 (C) 2001 年癌症研究运动。
Immunophototherapy of cancer combines the specificity of a monoclonal antibody (MAb) to an overexpressed tumor marker with the phototoxic properties of the conjugated dye. To analyze the potential role of internalisation of the dye on photo-induced cytotoxicity. we compared two target antigens, carcinoembryonic antigen (CEA) that does not internalise and ErbB2 that does. Human ovarian carcinoma SKOv3 cells that express a high level of ErbB2 were transfected with the CEA cDNA. Using FACS analysis, the resulting cell line, SKOv3-CEA-1B9, demonstrated comparable levels of expression of the two target antigens. Aluminium tetrasulfophthalocyanine (AlPcS4) was covalently coupled to anti-CEA MAb 35A7, anti-ErbB2 MAb FSP77 and a non-specific MAb PX, via a five-carbon sulfonamide spacer chain (A(1)) at molar ratios ranging from 6 to 9 moles of AlPcS4 per mole of MAb. The 35A7-(AlPcS(4)A(1))(8) conjugate induced 68% growth inhibition of the SKOv3-CEA-1B9 cell line after a 20 h incubation at 2.50 mug/ml (based on AlPcS(4)A(1) content) following light exposure. However, the FSP77-(AlPcS(4)A(1))(6) conjugate gave a 51 % growth inhibition for an AlPCS(4)A(1) concentration as low as 0.04 mug/ml after the same incubation time and exposure to the same light dose. At a 1.25 mug/ml AlPcS(4)A(1) concentration, the FSP77-(AlPcS(4)A(1))(6), conjugate gave a 67% growth inhibition after an incubation time as short as 1 h, reaching a 96% inhibition after an 8 h incubation time. Using an unique cell line that expresses two different target antigens. we demonstrated a clear advantage of an internalising over a non-internalising MAb-dye conjugate in terms of phototoxic efficacy. In vivo evaluation of the photodynamic properties of the conjugates is in progress. (C) 2001 Cancer Research Campaign.