恶性黑色素瘤细胞光免疫治疗的研究

Photoimmunotherapy of Human Ocular Melanoma Cells with McAb-photosensitizer

目的:探讨光免疫治疗(PIT)能否选择性提高人眼黑色素瘤细胞的光毒作用。方法:单克隆抗体IG12连结光敏物质CMA,形成免疫连结物(McAb-CMA),以评价其对体外人眼黑色素瘤培养细胞OCM431的光免疫治疗作用,RPMI1846黑色素瘤细胞与单克隆抗体IG12无交叉反应,作为对照组。氩离子-染料激光波长654nm,连结1mm光导纤维,光斑大小35mm。靶细胞和对照细胞的存活率用MTT法测定。结果:靶细胞OCM431预先与McAb-CMA培养,激光照射剂量5～40J/cm2显示靶细胞存活率随着激光剂量增加而明显降低。在相同激光照射剂量下,增加McAb-CMA的剂量,细胞存活曲线亦随着降低,当McAb-CMA的量增加超过6.7μmol/L时,肿瘤的光免疫杀伤作用趋平缓。对照细胞(RPMI1846)给予相同的照射条件,在激光剂量40J/cm2时仍有74%±2.6%细胞存活。结论:这一结果显示单克隆抗体IG12PIT可选择性提高人眼黑色素瘤的光杀伤作用。

Objective: To detect whether tumor-associated monoclonal antibodies can be used to enhance photodestruction of human ocular melanoma cells. Methods: Monoclonal antibody IG12 conjugated photosensitizer chlorin e6 mono-ethylenediamine onoamide (CMA) and evaluated the effectiveness of this immunoconjugate (McAb-CMA) in the destruction of OCM431 human ocular melanoma cells in vitro. RPMI1846 melanoma cells were used as non-target cells. Argon pumped dye laser was turned to 654nm, coupled to a 1mm quartz fiber optic and spot size was 35mm. Both target and non-target cells were treated with IC or light alone, Cell survival was assessed by colorimetric assay tetrazolium salt MTT. Results:Target cell (OCM431) preincubated with IC and irradiated with 5-40J/cm2 showed light dose-dependent decrease in cell survival. Under same light dose, cell survival decrease by increase McAb-CMA dose from 5.2霱 to 6.7霱. Non-target cells (RPMI1846) showed much less phototoxicity; cell survival was 74