Department of Pathology, Qianfoshan Hospital Affiliated with Shandong University, Jinan, Shandong 250014, PR China.
Mol Med Rep. 2012 Jan;5(1):103-7. doi: 10.3892/mmr.2011.610. Epub 2011 Oct 3.
HIV-1 derived lentiviral vectors (LVs) have emerged as a powerful tool for gene delivery. hTERT is an ideal tumor-associated antigen with which to develop a potential dendritic cell (DC) vaccine. The purpose of this study was to construct a recombinant lentivirus vector of the hTERT peptide and to determine the hTERT-specific cytotoxic T lymphocyte response elicited by DCs transfected with hTERT-lentivirus vectors in vitro. LVs encoding the hTERT peptide were constructed, DCs from cord blood were prepared, their morphology was observed and phenotype was analyzed by flow cytometry. Lenti-hTERT was transfected into DCs to construct the DC vaccines. T lymphocytes stimulated with DC vaccines and HepG2 cells (hTERT+) or 293T cells (hTERT-) were co-cultured for 24 h, respectively. The ability to stimulate proliferation of allogeneic T lymphocytes and the killing activity of CTLs activated by these DCs were determined using the MTT method. According to our results, the recombinant vector lenti-hTERT and lenti-hTERT-DC vaccine were successfully constructed. The stimulatory capacity of the lenti-hTERT DCs in the allogeneic T lymphocyte reaction was markedly enhanced compared with the DC control group (P<0.01). Inhibition rates in HepG2 cells of CTLs stimulated with lenti-hTERT-DCs (CTLT) were significantly higher than CTLs stimulated with the control DC group (CTLN) (P<0.01). Inhibition rates in 293T cells of CTLT and CTLN were low and there was no difference between the different DC groups (P>0.05). DCs transfected with the hTERT peptide were capable of eliciting a stronger hTERT-specific CTL response in vitro. Our data indicate that lenti-hTERT-DCs may potentially be used as an effective approach for cancer immunotherapy.
HIV-1 衍生的慢病毒载体（LVs）已成为基因递送的强大工具。hTERT 是一种理想的肿瘤相关抗原，可用于开发潜在的树突状细胞（DC）疫苗。本研究的目的是构建 hTERT 肽的重组慢病毒载体，并确定转染 hTERT-慢病毒载体的 DC 体外诱导的 hTERT 特异性细胞毒性 T 淋巴细胞反应。构建了编码 hTERT 肽的 LVs，制备了来自脐血的 DC，通过流式细胞术观察其形态并分析其表型。将 Lenti-hTERT 转染到 DC 中构建 DC 疫苗。用 DC 疫苗刺激 T 淋巴细胞，并分别与 HepG2 细胞（hTERT+）或 293T 细胞（hTERT-）共培养 24 小时。使用 MTT 法测定这些 DC 激活的同种异体 T 淋巴细胞增殖能力和 CTL 的杀伤活性。根据我们的结果，成功构建了重组载体 lenti-hTERT 和 lenti-hTERT-DC 疫苗。与 DC 对照组相比，lenti-hTERT-DC 对同种异体 T 淋巴细胞反应的刺激能力明显增强（P<0.01）。用 lenti-hTERT-DC 刺激的 CTL（CTLT）对 HepG2 细胞的抑制率明显高于用对照 DC 组刺激的 CTL（CTLN）（P<0.01）。CTLT 和 CTLN 对 293T 细胞的抑制率较低，不同 DC 组之间无差异（P>0.05）。转染 hTERT 肽的 DC 能够在体外引发更强的 hTERT 特异性 CTL 反应。我们的数据表明，lenti-hTERT-DC 可能是癌症免疫治疗的一种有效方法。
