Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
Vaccine. 2012 May 14;30(23):3395-404. doi: 10.1016/j.vaccine.2012.03.045. Epub 2012 Apr 3.
Human telomerase reverse transcriptase (hTERT) has been identified as an ideal tumor-associated antigen (TAA). Use of a synthetic hTERT epitope peptide to pulse dendritic cells can induce autologous T cell anti-tumor immune responses, but such responses induced by a single epitope peptide have been shown to be weak and a narrow-spectrum. Here, we designed dendritic tandem multiple antigenic peptides (MAPs) containing the following three hTERT epitope peptides: I540, V461 and L766, which are HLA-A02-, HLA-A24- and HLA-RDB104/11/15-restricted, respectively. The MAPs and their three single-epitope peptides were obtained through solid-phase synthesis. Healthy volunteers that were HLA-A02(+)/HLA-DRB104(+) and HLA-A24(+)/HLA-DRB115(+) were recruited. Myeloid dendritic cells were isolated by magnetic activated cell sorting and were divided into a MAP-stimulated group (MAP-DC), a group in which the three epitope peptides were mixed and used to stimulate the DCs (MixP-DC) and a no peptide-stimulated group (NoP-DC, control group). All of the DCs were cultured in serum-free medium, pulsed with the corresponding peptides on the 3rd, 5th and 7th days, and co-cultured with autologous lymphocytes when they were mature. The related cytokines were measured via ELISA. The killing effects of cytotoxic T lymphocytes (CTLs) on SW480/A549 tumor cells expressing HLA-A02(+), HepG2/SMMC-7721 cells expressing HLA-A24(+) and SKOV3 cells negative for HLA-A02/A*24 were detected by flow cytometry. Our results indicated that the CTLs induced by the MAP-DCs had the greatest anti-tumor effect. Therefore, the dendritic tandem multiple antigenic hTERT epitope peptides combined with MDCs may represent a powerful, broad-spectrum anti-tumor vaccine.
人类端粒酶逆转录酶（hTERT）已被鉴定为理想的肿瘤相关抗原（TAA）。使用合成的 hTERT 表位肽脉冲树突状细胞可以诱导自体 T 细胞抗肿瘤免疫反应，但已证明由单个表位肽诱导的此类反应较弱且谱较窄。在这里，我们设计了包含以下三个 hTERT 表位肽的树突状串联多抗原肽（MAP）：I540、V461 和 L766，它们分别受 HLA-A02-、HLA-A24- 和 HLA-RDB104/11/15 限制。MAP 及其三个单表位肽通过固相合成获得。招募了 HLA-A02(+)/HLA-DRB104(+) 和 HLA-A24(+)/HLA-DRB115(+) 的健康志愿者。通过磁激活细胞分选分离髓样树突状细胞，并将其分为 MAP 刺激组（MAP-DC）、三种表位肽混合刺激 DC 的组（MixP-DC）和无肽刺激组（NoP-DC，对照组）。所有 DC 在无血清培养基中培养，在第 3、5 和 7 天用相应的肽脉冲，成熟时与自体淋巴细胞共培养。通过 ELISA 测量相关细胞因子。通过流式细胞术检测表达 HLA-A02(+) 的 SW480/A549 肿瘤细胞、表达 HLA-A24(+) 的 HepG2/SMMC-7721 细胞和 HLA-A02/A*24 阴性的 SKOV3 细胞的细胞毒性 T 淋巴细胞（CTL）对的杀伤作用。结果表明，MAP-DC 诱导的 CTL 具有最强的抗肿瘤作用。因此，与 MDC 结合的树突状串联多抗原 hTERT 表位肽可能代表一种强大的广谱抗肿瘤疫苗。
