抗HPV E7小鼠单克隆抗体与人类宫颈癌细胞的特异性结合

使用抗HPV E7蛋白的小鼠单克隆抗体进行Western blot杂交，发现3种人类宫颈癌细胞系CaSki、Hela S3和SiHa均表达一定量的HPV E7病毒蛋白，表达量由高至低依次为CaSki，Hela S3，SiHa．选取CaSki为代表，用荧光免疫标记法检测抗E7抗体分子与活体癌细胞中HPV E7蛋白结合的情况．结果发现，仅衰亡细胞、0．3％Triton X-100处理30min后的活细胞以及^137 Cs辐射处理后的活细胞显示荧光，暗示只有细胞膜通透性发生改变后抗E7小鼠单克隆抗体分子才能进入细胞与HPV E7蛋白特异地结合．流式细胞仪检测表明，与同型IgG抗体mAbS26相比，抗E7抗体分子与CaSki细胞作用时荧光标记细胞的净增百分率为16．5％，说明抗E7小鼠单克隆抗体分子与CaSki细胞结合具有特异性．

Anti-HPV E7 Mouse Monoclonal Antibody Specifically Targets the HPV-Associated Cervical Cancers Cells

Western blot showed that human cervical cancer cell lines CaSki, Hela S3 and SiHa were capable of expressing certain amount of HPV E7 proteins when anti-HPV E7 mAb was used. The amount of E7 protein in CaSki was higher as compared with that in Hela S3 or SiHa. SiHa showed much less amount of E7 protein among three cell lines. Hence, CaSki was chosen for further investigation in order to understand whether the anti-HPV E7 antibody interacted with those cervical cancer cells containing HPV DNA. The investigation by immunofluorescence microscopy revealed that the anti-HPV E7 mAb was unable to interact with those cells that were thriving, but targeted those cells that were dying or damaged by Triton X-100 treatment and 137Cs irradiation. Flow cytometry gave a net-increased value of 16.5% to the interaction between the anti-HPV E7 mAb and CaSki cells as compared with that for the isotype HBsAg antibody. High percentage of cells binding to the FITC-conjugated antibody indicated the specific interaction between the anti-HPV E7 mAb and the E7 protein inside CaSki cells.