腺病毒载体编码人甲胎蛋白与γ干扰素对小鼠肝癌免疫的协同作用

[Synergic effect of adenoviral vector-encoding human alpha-fetoprotein and interferon-gamma on immunity against hepatocellular carcinoma in mice].

作者信息

Tan Xiao-Hua, Zhu Qing

机构信息

Center for Gene Therapy, Department of Molecular and Pathological Medicine, Medical Center, McMaster University, Hamilton, ON. Canada, L8S.

出版信息

Ai Zheng. 2008 Feb;27(2):155-9.

PMID:18279612

Abstract

BACKGROUND & OBJECTIVE: Primary hepatocellular carcinoma (HCC) is a common malignant tumor. Up to date, no effective treatment for HCC is available. This study was to investigate the synergic effect of adenoviral vector-encoding xenogeneic alpha-fetoprotein (AFP) and interferon-gamma (IFN-gamma) on the immunity against HCC in mice.

METHODS

IFN-gamma gene was cloned using reverse transcription-polymerase chain reaction (RT-PCR). Replication-defective adenovirus encoding both human AFP and murine IFN-gamma was constructed. The cytotoxic activity of antigen-specific cytotoxic T lymphocytes (CTLs) was detected 7 days after the intradermal immunization of C57BL/6 mice with Ad-hAFP, Ad-IFN-gamma or Ad-hAFP/IFN-gamma using standard (51)Cr release assay. The survival of mice after the subcutaneous inoculation of 5x10(6) hepatoma Hepa 1-6 cells was checked.

RESULTS

The cytotoxic activity of CTLs elicited by Ad-hAFP/IFN-gamma was much stronger than that by Ad-hAFP or Ad-IFN-gamma alone. At an effector:target (E:T) ratio of 10:1, the cytotoxic activities of CTLs in Ad-hAFP/IFN-gamma, Ad-hAFP, and Ad-IFN-gamma groups were (43.8+/-5.5)%, (28.2+/-3.2)%, and (12.8+/-1.9)%, at a ratio of 30:1, were (79.6+/-6.4)%, (51.9+/-4.3)%, and (15.6+/-2.3)%, and at a ratio of 90:1, were (88.2+/-6.3)%, (62.5+/-4.8)%, and (26.5+/-2.4)%, respectively. The tumor formation rates were 80% in Ad-hAFP-immunized mice and 100% in Ad-IFN-gamma-immunized mice at 2 months after inoculation with Hepa1-6 cells; no tumor formed in Ad-hAFP/IFN-gamma-immunized mice, and all mice in this group survived during two-month observation.

CONCLUSION

Ad-hAFP can efficiently induce immunity against HCC in mice, and IFN-gamma enhances such an effect.

摘要

背景与目的

原发性肝细胞癌（HCC）是一种常见的恶性肿瘤。迄今为止，尚无有效的HCC治疗方法。本研究旨在探讨编码异种甲胎蛋白（AFP）和干扰素-γ（IFN-γ）的腺病毒载体对小鼠抗HCC免疫的协同作用。

方法

采用逆转录-聚合酶链反应（RT-PCR）克隆IFN-γ基因。构建编码人AFP和鼠IFN-γ的复制缺陷型腺病毒。用标准的（51）Cr释放试验，在C57BL/6小鼠皮内注射Ad-hAFP、Ad-IFN-γ或Ad-hAFP/IFN-γ 7天后，检测抗原特异性细胞毒性T淋巴细胞（CTL）的细胞毒活性。检查皮下接种5×10（6）个肝癌Hepa 1-6细胞后小鼠的存活情况。

结果

Ad-hAFP/IFN-γ诱导的CTL细胞毒活性比单独的Ad-hAFP或Ad-IFN-γ强得多。在效应细胞与靶细胞（E:T）比例为10:1时，Ad-hAFP/IFN-γ、Ad-hAFP和Ad-IFN-γ组CTL的细胞毒活性分别为（43.8±5.5）%、（28.2±3.2）%和（12.8±1.9）%；在比例为30:1时，分别为（79.6±6.4）%、（51.9±4.3）%和（15.6±2.3）%；在比例为90:1时，分别为（88.2±6.3）%、（62.5±4.8）%和（26.5±2.4）%。接种Hepa1-6细胞2个月后，Ad-hAFP免疫小鼠的肿瘤形成率为80%，Ad-IFN-γ免疫小鼠的肿瘤形成率为100%；Ad-hAFP/IFN-γ免疫小鼠未形成肿瘤，该组所有小鼠在两个月的观察期内均存活。