[抗独特型微型抗体脉冲树突状细胞诱导针对自体卵巢癌的T细胞反应]

[Induction of T cell responses against autologous ovarian cancer by anti-idiotype minibody-pulsed dendritic cells].

作者信息

Yang Wen-Lan, Cui Heng, Feng Jie, Chang Xiao-Hong, Fu Tian-Yun, Ye Xue, Zhang Hong, Li Xiao-Ping, Wen Hong-Wu, Feng Li-Min, Tong Chun-Rong

机构信息

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

出版信息

Ai Zheng. 2004 Dec;23(12):1639-45.

PMID:15601552

Abstract

BACKGROUND & OBJECTIVE: Immunotherapy of sensitizing dendritic cells (DCs) with antigen,protein,and frozen cancer cell has been widely used in treating various cancers. The 6B11 anti-idiotype-antibody,a fusion protein prepared by our research center,can mimic ovarian cancer-associated antigen OC166-9. This study was to induce T cell cytotoxicity against autologous tumor cells of patients with ovarian cancer by 6B11 anti-idiotype-antibody.

METHODS

Peripheral blood samples were collected from 10 patients with epithelial ovarian cancer,Monocytes were isolated and cultured to obtain DCs. Immature DCs were stimulated with 6B11 anti-idiotype-antibody (MINI-DC group); unpulsed DCs (unpulsed-DC group),mouse F(ab) '2 fragments pulsed DCs [F(ab)'2-DC group],and T cells alone (T group) were served as controls. Mature DCs were harvested. (3)H-thymidine ((3)H-TdR) incorporation approach was used to measure effect of DCs on stimulating auto-T cell proliferation. Cytotoxicity of DC-activated T cells against auto-tumor cells was measured with (51)Cr 6-h release test,tumor cell lines,SKOV3,HLE,and K562, were used as controls.

RESULTS

In 4 cases,cpm value of (3)H-TdR incorporation,as symbol of auto-T cell proliferation, in MINI-DC group was significantly higher than those in control groups. In 5 cases,specific cytotoxicity effect of T cells on auto-tumor cells was observed in MINI-DC group at effect-target ratio of 20:1,the toxicity effect of T cells in MINI-DC group was 25%-100%,significantly higher than those in F(ab)'2-DC group (18%-40%), unpulsed-DC group (13%-43%),and T group (9%-58%). In 4 cases,the toxicity effect of T cells in MINI-DC group, at effect-target ratio of 20:1,on auto-tumor cells was 25%-100%, higher than those on SKOV3 cells (5%-51%),HLE cells (2%-38%),and K562 cells (2%-25%). Moreover,the toxicity effect of T cells in MINI-DC group on auto-tumor cells can be partially blocked by anti-MHC-I antibody,which indicated that the toxicity was antigen-specific.

CONCLUSION

DCs loaded with 6B11 anti-idiotype antibody that mimic ovarian cancer antigen can induce antigen specific T cell cytotoxicity against auto-ovarian tumor cells in vitro.

摘要

背景与目的

用抗原、蛋白质及冻存癌细胞致敏树突状细胞（DCs）进行免疫治疗已广泛应用于多种癌症的治疗。本研究中心制备的融合蛋白6B11抗独特型抗体可模拟卵巢癌相关抗原OC166 - 9。本研究旨在通过6B11抗独特型抗体诱导卵巢癌患者自体肿瘤细胞的T细胞细胞毒性。

方法

采集10例上皮性卵巢癌患者的外周血样本，分离培养单核细胞以获得DCs。用6B11抗独特型抗体刺激未成熟DCs（MINI - DC组）；未致敏DCs（未致敏 - DC组）、小鼠F(ab)'2片段致敏DCs [F(ab)'2 - DC组]及单独的T细胞（T组）作为对照。收获成熟DCs。采用³H - 胸腺嘧啶核苷（³H - TdR）掺入法检测DCs对自体T细胞增殖的刺激作用。用⁵¹Cr 6小时释放试验检测DC激活的T细胞对自体肿瘤细胞的细胞毒性，以肿瘤细胞系SKOV3、HLE及K562作为对照。

结果

4例患者中，以³H - TdR掺入的cpm值作为自体T细胞增殖的标志，MINI - DC组显著高于对照组。5例患者中，在效靶比为20∶1时，MINI - DC组观察到T细胞对自体肿瘤细胞的特异性细胞毒性作用，MINI - DC组T细胞的毒性作用为25% - 100%，显著高于F(ab)'2 - DC组（18% - 40%）、未致敏 - DC组（13% - 43%）及T组（9% - 58%）。4例患者中，在效靶比为20∶1时，MINI - DC组T细胞对自体肿瘤细胞的毒性作用为25% - 100%，高于对SKOV3细胞（5% - 51%）、HLE细胞（2% - 38%）及K562细胞（2% - 25%）的毒性作用。此外，MINI - DC组T细胞对自体肿瘤细胞的毒性作用可被抗MHC - I抗体部分阻断，提示该毒性具有抗原特异性。