Chang Xiao-hong, Ye Xue, Cui Heng, Feng Jie, Li Yi, Zhu Hong-lan, Yang Wen-lan, Fu Tian-yun, Cheng Hong-yan, Guo Hui-fang
Gynecologic Oncology Center, Peking University People's Hospital, Beijing 100044, China.
Chin Med J (Engl). 2006 Jan 20;119(2):131-9.
We have previously developed and characterized a monoclonal anti-idiotype antibody, designated 6B11, which mimics an ovarian carcinoma associated antigen OC166 - 9 and whose corresponding monoclonal antibody is COC166 - 9 (Ab1). In this study, we evaluate the humoral immune responses induced by the fusion protein 6B11 single-chain variable fragment (scFv)/human granulocyte macrophage colony-stimulating factor (hGM-CSF) and 6B11scFv in BALB/c mice.
The fusion protein 6B11scFv/hGM-CSF was constructed by fusing a recombinant single-chain variable fragment of 6B11scFv to GM-CSF. BALB/c mice were administrated by 6B11scFv/hGM-CSF and 6B11scFv, respectively.
The fusion protein 6B11scFv/hGM-CSF retained binding to the anti-mouse F (ab) 2' and was also biologically active as measured by proliferation of human GM-CSF dependent cell TF1 in vitro. After immunization with the 6B11scFv/hGM-CSF and 6B11ScFv, BALB/c mice showed significantly enhanced Ab3 antibody responses to 6B11scFv/hGM-CSF compared with the 6B11scFv alone. The level of Ab3 was the highest after the first week and maintained for five weeks after the last immunization. Another booster was given when the Ab3 titer descended, and it would reach to the high level in a week.
The fusion protein 6B11scFv/hGM-CSF can induce humoral immunity against ovarian carcinoma in vivo. We also provide the theoretical foundation for the application of the fusion protein 6B11scFv/hGM-CSF for active immunotherapy of ovarian cancer.
我们先前已研发并鉴定出一种单克隆抗独特型抗体,命名为6B11,它模拟一种卵巢癌相关抗原OC166 - 9,其对应的单克隆抗体为COC166 - 9(Ab1)。在本研究中,我们评估融合蛋白6B11单链可变片段(scFv)/人粒细胞巨噬细胞集落刺激因子(hGM - CSF)和6B11scFv在BALB/c小鼠中诱导的体液免疫反应。
通过将6B11scFv的重组单链可变片段与GM - CSF融合构建融合蛋白6B11scFv/hGM - CSF。分别用6B11scFv/hGM - CSF和6B11scFv对BALB/c小鼠进行给药。
融合蛋白6B11scFv/hGM - CSF保留了与抗小鼠F(ab)2'的结合能力,并且通过人GM - CSF依赖细胞TF1的体外增殖测定显示具有生物活性。用6B11scFv/hGM - CSF和6B11ScFv免疫后,与单独使用6B11scFv相比,BALB/c小鼠对6B11scFv/hGM - CSF的Ab3抗体反应显著增强。Ab3水平在第一周后最高,并在最后一次免疫后维持五周。当Ab3滴度下降时给予另一次加强免疫,一周内它会达到高水平。
融合蛋白6B11scFv/hGM - CSF可在体内诱导针对卵巢癌的体液免疫。我们还为融合蛋白6B11scFv/hGM - CSF用于卵巢癌主动免疫治疗的应用提供了理论基础。