人肝癌靶向脂质体微泡造影剂的鉴定及免疫学性质研究

目的 制备人肝细胞癌靶向免疫脂质体微泡造影剂，并对其免疫学性质进行研究。方法 静电吸附法制备免疫脂质体微泡造影剂；玻片凝集试验、荧光免疫染色试验证明抗体与脂质体微泡的结合；ELISA法检测免疫脂质体微泡的活性；花环形成试验、花环形成阻断试验及荧光免疫染色试验研究免疫脂质体微泡的靶向作用；细胞毒试验(MTT法)评价其安全性。结果 免疫脂质体微泡的玻片凝集试验、荧光免疫染色试验均为阳性；ELISA结果显示，免疫脂质体微泡中单抗的活性与游离单抗基本一致；免疫脂质体微泡围绕人肝癌细胞形成花环，花环形成率达90％以上；免疫脂质体微泡造影剂对人正常肝细胞的增殖没有影响。结论 采用静电吸附法成功制备了具有良好免疫活性的人肝细胞癌靶向免疫脂质体造影剂；该造影剂在体外能与人肝癌细胞高效特异结合，对人正常肝细胞没有毒性。

Study on immunological properties of human hepatocellular carcinoma-targeted liposome microbubbles

Objective To prepare human hepatocellular carcinoma targeted liposome microbubbles and investigate their immunological properties.Methods Human hepatocarcinoma specific monoclonal antibody HAb18 was attached to the surface of self made liposome microbubbles by electrostatic attraction to prepare immunoliposome microbubbles.The combination of HAb18 with liposome microbubbles was proved by slide agglutination test and immunofluorescent assay.Their immunological activity was measured by ELISA.Rosset formation test,rosset formation blocking test and immunofluorescent assay were used to identify the specific binding of immunoliposome microbubbles to SMMC 7721 cells,while cytotoxicity assay was used to detect their effect on human hepatocytes.Results Immunoliposome microbubbles were positive in slide agglutination test and immunofluorescent assay.ELISA indicated that the immunological activity of HAb18 on liposome microbubbles was similar to that of free HAb18.SMMC 7721 cells were surrounded by immunoliposome microbubbles to form rossets,while control cells were negative.Proliferation of human hepatocytes was not influenced by immunoliposome microbubbles.Conclusions Immunoliposome microbubbles with high specific biological activity were prepared successfully.The contrast agent can bind to human hepatocarcinoma cells specially and effectively.The results may pave the way for further studies.