Department of Biochemistry, Faculty of Pharmacy, Tanta University, El-Bahr Street, Tanta, 31527, El-Gharbiya, Egypt.
Department of Clinical Pathology, Faculty of Medicine, Tanta University, Tanta, Egypt.
Clin Transl Oncol. 2019 May;21(5):636-645. doi: 10.1007/s12094-018-1965-6. Epub 2018 Oct 27.
Hepatocellular carcinoma (HCC) is one of the common malignancies, characterized by poor response to conventional therapeutic options. Immunotherapy with dendritic cells (DCs)-vaccines is one of the most successful strategies used for the treatment of HCC. However, the methods applied in the preparation of antigen-loaded DCs are important factors for optimization of DCs vaccines.
The present study was conducted to investigate the effect of HCC-whole tumor cell lysate prepared using rapid repetitive freeze-thaw cycles on the immunogenicity of DCs and evaluate the ability of whole tumor cell lysate-pulsed DCs vaccine to induce a specific cytotoxic T lymphocytes (CTLs) response against HepG2 cell line.
Immature DCs generated from peripheral blood monocytes were randomized into two groups: control DCs and whole tumor cell lysate-pulsed DCs. Phenotypic analysis of the DCs' cell maturation marker CD83 and co-stimulatory molecule CD86 was performed. HCC-specific cytotoxic activity of CD8 CTLs was measured in vitro.
Loading of DCs with necrotic whole cell lysate resulted in non-significant changes in DCs' expression of CD83, but a significant increase in expression of CD86. In addition, CD8 CTLs stimulated with whole tumor cell lysate-pulsed DCs showed a high cytotoxic activity that specifically attack HepG2 cells.
Our findings indicated that pulsation of DCs with whole tumor cell lysate prepared by repetitive freeze-thaw cycles could efficiently enhance the ability of DCs to induce proliferation and clonal expansion of CD8 CTLs. Data herein, also indicated that whole tumor cell lysate-pulsed DCs triggers a specific CD8 CTLs against HCC tumor cells.
肝细胞癌（HCC）是常见的恶性肿瘤之一，其对常规治疗方案的反应较差。树突状细胞（DC）疫苗的免疫疗法是治疗 HCC 最成功的策略之一。然而，用于制备负载抗原的 DC 的方法是优化 DC 疫苗的重要因素。
本研究旨在探讨使用快速重复冻融循环制备的 HCC 全肿瘤细胞裂解物对 DC 免疫原性的影响，并评估全肿瘤细胞裂解物脉冲 DC 疫苗诱导针对 HepG2 细胞系的特异性细胞毒性 T 淋巴细胞（CTLs）反应的能力。
从外周血单核细胞中生成未成熟的 DC，随机分为两组：对照组 DC 和全肿瘤细胞裂解物脉冲 DC。对 DC 细胞成熟标志物 CD83 和共刺激分子 CD86 的表型分析。在体外测量 CD8 CTLs 对 HCC 的特异性细胞毒性活性。
用坏死全细胞裂解物负载 DC 不会导致 DC 表达 CD83 的显著变化，但会导致 CD86 的表达显著增加。此外，用全肿瘤细胞裂解物脉冲的 DC 刺激的 CD8 CTLs 显示出针对 HepG2 细胞的高细胞毒性活性。
我们的研究结果表明，用重复冻融循环制备的全肿瘤细胞裂解物脉冲 DC 可以有效地增强 DC 诱导 CD8 CTLs 增殖和克隆扩增的能力。本文的数据还表明，全肿瘤细胞裂解物脉冲的 DC 可引发针对 HCC 肿瘤细胞的特异性 CD8 CTLs。
