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[目的]制备小鼠抗人IgG-葡聚糖—盐酸吡柔比星免疫靶向偶联物,在肿瘤组织中接种人IgG非特异性靶点,探讨人造靶点免疫导向治疗肿瘤的可行性。[方法]制备小鼠抗人IgG-葡聚糖—盐酸吡柔比星(THP)偶联物,建立荷瘤小鼠模型。采用ELISA法测定小鼠抗人IgG-葡聚糖—盐酸吡柔比星偶联物抗体活性;体外细胞毒性实验(四甲基偶氮唑蓝法)比较分析小鼠抗人IgG、游离盐酸吡柔比星、小鼠抗人IgG-葡聚糖—盐酸吡柔比星对S180细胞的毒性作用;采用高效液相法(HPLC)测定肿瘤组织中盐酸吡柔比星药物浓度。[结果]制备的偶联物中,小鼠抗人IgG、葡聚糖与盐酸吡柔比星物质的量比为1∶2.5∶54;偶联物保留了小鼠抗人IgG抗体活性;体外细胞毒性实验证明偶联物对S180细胞有体外杀伤作用。对荷S180肉瘤小鼠进行体内导向治疗试验,高效液相色谱法测定小鼠抗人IgG-葡聚糖—盐酸吡柔比星+人IgG组中肿瘤组织含盐酸吡柔比星含量较盐酸吡柔比星组增加(P<0.01)。[结论]通过在肿瘤组织中接种IgG非特异性靶点,用抗人IgG单克隆免疫导向吡柔比星治疗小鼠S180肉瘤具有一定的可行性。
[Objective] The aim of the study was to prepare mouse anti-human IgG-dextran-pirarubicin immunoconjugate conjugate, inoculate human non-specific target IgG in tumor tissue and explore the feasibility of using artificial target-directed immunotherapy to treat tumor. [Method] The mouse anti-human IgG-dextran-pirarubicin hydrochloride (THP) conjugate was prepared and the tumor-bearing mouse model was established. Antibody activity of mouse anti-human IgG-dextran-pirarubicin conjugate was determined by ELISA. In vitro cytotoxicity assay (MTT assay) was used to compare the anti-human IgG, free hydrochloric acid Epirubicin and mouse anti-human IgG-dextran-pirarubicin on S180 cells. The concentration of pirarubicin hydrochloride in tumor tissue was determined by HPLC. [Result] In the prepared conjugate, the ratio of mouse anti-human IgG, dextran and pirarubicin hydrochloride was 1: 2.5: 54; the conjugate retained the activity of mouse anti-human IgG antibody; Cytotoxicity experiments show that the conjugate has the killing effect on S180 cells in vitro. The S180 sarcoma mice were treated with the in vivo targeting test. The contents of piruettes in the tumor tissue in the anti-human IgG-dextran-pirarubicin hydrochloride + human IgG group were determined by HPLC compared with that of the hydrochloride Rousuyin group increased (P <0.01). [Conclusion] It is feasible to immunize mice S180 sarcoma with anti-human IgG monoclonal antibody to pirarubicin by inoculating nonspecific target of IgG in tumor tissue.