融合蛋白6B11脉冲树突状细胞诱导的T细胞对卵巢癌细胞的细胞毒性作用。

Cytotoxic effects of T cells induced by fusion protein 6B11-pulsed dendritic cells on ovarian carcinoma cells.

作者信息

Yang Wenlan, Feng Jie, Chang Xiaohong, Fu Tianyun, Ye Xue, Zhang Hong, Li Xiaoping, Wen Hongwu, Feng Limin, Tong Chunrong, Cui Heng

机构信息

Gynecologic Oncology Center, People's Hospital, Peking University, Beijing, 100044, PR China.

出版信息

Gynecol Oncol. 2007 Apr;105(1):238-43. doi: 10.1016/j.ygyno.2006.04.028.

DOI:10.1016/j.ygyno.2006.04.028

PMID:17383546

Abstract

INTRODUCTION

6B11 anti-idiotype minibody, a fusion protein, has been shown to mimic ovarian carcinoma associated antigen OC166-9. This study was designed to determine whether 6B11 anti-idiotype minibody-pulsed dendritic cells (DCs) can induce cytotoxic T cells against ovarian cancer cells.

METHODS

Monocytes were isolated from peripheral blood mononuclear cells collected from patients with epithelial ovarian carcinoma (n=10). The monocytes-derived immature DCs were stimulated by cytokines, and mature DCs were pulsed with 6B11 anti-idiotype-minibody or murine F(ab)'2 fragments. The proliferation of autologous T cells induced by DCs was determined by 3H-thymidine uptake. The cytotoxicity of DC-activated T cells against autologous carcinoma cells was determined by 51Cr-release assay.

RESULTS

Purified T cells demonstrated strong proliferation following incubation with 6B11 anti-idiotype minibody-pulsed DCs in 4 of 10 patients. The specific cytotoxicity of purified T cells against autologous carcinoma cells was induced after stimulation with 6B11 anti-idiotype minibody-pulsed DCs in 5 of 10 patients with cytotoxic effects ranging from 25 to 95%. In contrast, isotype murine F(ab)'2 fragments-pulsed DCs did not induce T cell proliferation and cytotoxicity against the targets. Additionally, the cytotoxic effect was partially inhibited by anti-MHC class-I antibody indicating that the cytotoxic effects are antigen-specific.

CONCLUSION

6B11 anti-idiotype-antibody-pulsed DCs can induce T cell proliferation and T cell-mediated cytotoxicity against autologous ovarian tumor cells in vitro. The cytotoxic effects of T cells against autologous tumor cells are antigen-specific. These data implicate the rationale for the use of 6B11 anti-idiotype minibody as immunotherapy against ovarian carcinoma.

摘要

引言

6B11抗独特型微抗体是一种融合蛋白，已被证明可模拟卵巢癌相关抗原OC166 - 9。本研究旨在确定6B11抗独特型微抗体脉冲树突状细胞（DCs）是否能诱导针对卵巢癌细胞的细胞毒性T细胞。

方法

从上皮性卵巢癌患者（n = 10）采集的外周血单个核细胞中分离单核细胞。单核细胞来源的未成熟DCs用细胞因子刺激，成熟DCs用6B11抗独特型微抗体或鼠F(ab)'2片段脉冲。通过3H - 胸腺嘧啶核苷摄取测定DCs诱导的自体T细胞增殖。通过51Cr释放试验测定DC激活的T细胞对自体癌细胞的细胞毒性。

结果

在10例患者中的4例中，纯化的T细胞与6B11抗独特型微抗体脉冲的DCs孵育后显示出强烈增殖。在10例患者中的5例中，用6B11抗独特型微抗体脉冲的DCs刺激后诱导了纯化的T细胞对自体癌细胞的特异性细胞毒性，细胞毒性作用范围为25%至95%。相比之下，同型鼠F(ab)'2片段脉冲的DCs未诱导T细胞增殖和对靶标的细胞毒性。此外，抗MHC I类抗体部分抑制了细胞毒性作用，表明细胞毒性作用是抗原特异性的。