Department of Pathology, Fujian Medical University, 88# Jiao Tong Road, Fuzhou, 350004, Fujian, People's Republic of China.
J Cancer Res Clin Oncol. 2014 Feb;140(2):281-9. doi: 10.1007/s00432-013-1552-8. Epub 2013 Dec 10.
Melanoma antigen gene A3 (MAGE-A3) is aberrantly expressed in a number of cancer types. Because of its high specificity, MAGE-A3 has shown to be a promising candidate for cancer immunotherapy. Dendritic cells (DCs) have emerged as the natural agents for antigen delivery. DCs transduced with antigen may increase immune response and maintain immune durability. The aim of this study was to investigate the roles of DCs transduced with lentiviral vectors (LVs) encoding full-length MAGE-A3 gene in cancer immunotherapy .
A LV containing full-length MAGE-A3 gene (rLV/MAGE-A3) was constructed. Reverse transcriptase-polymerase chain reaction and direct DNA sequencing were performed to verify the construct. Human DCs derived from umbilical cord blood were then transduced with rLV/MAGE-A3. The potency of rLV/MAGE-A3-transduced DCs was examined by measurement of surface markers and mixed lymphocyte reaction. The MAGE-A3-specific T-cell response induced by DCs was detected using the lactate dehydrogenase release assay.
rLV/MAGE-A3 was constructed successfully and used to transduce DCs efficiently. DCs transduced with rLV/MAGE-A3 stably expressed MAGE-A3 and yielded high percentage of cells expressing CD80, CD86, and HLA-DR. rLV/MAGE-A3 transduction did not impair DCs viability and maturation at a multiplicity of infection of 30. The rLV/MAGE-A3-transduced DCs induced MAGE-A3-specific T lymphocytes that exhibited a significant lysis activity against MAGE-A3-bearing tumor cell lines (HuH-7 and SGC-7901).
DC-directed rLV/MAGE-A3 efficiently induced antigen-specific immune responses, indicating the possibility of DC-based MAGE-A3 antigen vaccine as a promising strategy for treatment of MAGE-A3-associated cancer.
黑色素瘤相关抗原基因 A3（MAGE-A3）在多种癌症类型中异常表达。由于其高度特异性，MAGE-A3 已被证明是癌症免疫治疗的有前途的候选物。树突状细胞（DCs）已成为抗原递呈的天然剂。转导抗原的 DC 可以增强免疫反应并保持免疫持久性。本研究旨在探讨转导全长 MAGE-A3 基因的慢病毒载体（LVs）的 DC 在癌症免疫治疗中的作用。
构建含有全长 MAGE-A3 基因的 LV（rLV/MAGE-A3）。进行逆转录-聚合酶链反应和直接 DNA 测序以验证构建。然后用人脐带血来源的 DC 转导 rLV/MAGE-A3。通过测量表面标志物和混合淋巴细胞反应来检查 rLV/MAGE-A3 转导的 DC 的效力。通过乳酸脱氢酶释放测定法检测由 DC 诱导的 MAGE-A3 特异性 T 细胞反应。
成功构建了 rLV/MAGE-A3，并有效地用于转导 DC。rLV/MAGE-A3 转导的 DC 稳定表达 MAGE-A3，并产生高比例表达 CD80、CD86 和 HLA-DR 的细胞。在感染复数为 30 时，rLV/MAGE-A3 转导不会损害 DC 的活力和成熟度。rLV/MAGE-A3 转导的 DC 诱导了 MAGE-A3 特异性 T 淋巴细胞，这些细胞对携带 MAGE-A3 的肿瘤细胞系（HuH-7 和 SGC-7901）表现出明显的裂解活性。
DC 定向的 rLV/MAGE-A3 有效地诱导了抗原特异性免疫反应，表明基于 DC 的 MAGE-A3 抗原疫苗作为治疗 MAGE-A3 相关癌症的有前途策略的可能性。
