Regresar

Internalisation enhances photo-induced cytotoxicity of monoclonal antibody-phthalocyanine conjugates

Abstract:

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 (AIPcS4) 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 (A1) at molar ratios ranging from 6 to 9 moles of AIPcS4 per mole of MAb. The 35A7-(AIPcS4A1)8 conjugate induced 68% growth inhibition of the SKOv3-CEA-1B9 cell line after a 20 h incubation at 2.50 μg/ml (based on AIPcS4A1 content) following light exposure. However, the FSP77-(AIPcS4A1)6 conjugate gave a 51% growth inhibition for an AIPcS4A1 concentration as low as 0.04 μg/ml after the same incubation time and exposure to the same light dose. At a 1.25 μg/ml AIPcS4A1 concentration, the FSP77-(AIPcS4A1)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. © 2001 Cancer Research Campaign.