Siriraj Center of Research Excellence for Cancer Immunotherapy, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Graduate Program in Immunology, Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Sci Rep. 2019 Nov 27;9(1):17668. doi: 10.1038/s41598-019-54087-0.
Cellular immunotherapy is a promising new therapeutic approach for hepatocellular carcinoma (HCC), which has a high recurrence rate, irrespective of the treatment administered. In this study, we attempted to improve the cytolytic activity of effector T-lymphocytes against HCC. T-lymphocytes were activated by monocyte-derived dendritic cells (DCs) pulsed with cell lysate or RNA prepared from HCC cell lines. Monocytes were activated for differentiation into DCs by treatment with the IL4 and GM-CSF. DCs were pulsed with cell lysate or RNA prepared from a single cell line or combinations of two or three HCC cell lines, and then co-cultured with autologous T-lymphocytes with the intent of creating specific cytotoxicity. We discovered that DCs pulsed with total RNA effectuated greater T-lymphocyte function than DCs pulsed with total cell lysate, as evidenced by greater cytolytic activities against HCC target cells. The percentage of Huh7, HepG2, and SNU449 cell apoptosis at effector:target ratio of 10:1 was 42.6 ± 4.5% (p = 0.01), 33.6 ± 3.1% (p = 0.007), and 21.4 ± 1.4% (p < 0.001), respectively. DCs pulsed with pools of antigens prepared from three cell lines improved the cytolytic function of effector T-lymphocytes by approximately two-fold (p < 0.001), which suggests that this approach be further developed and applied for adoptive transfer treatment of HCC.
细胞免疫疗法是一种有前途的新治疗方法，可用于治疗肝细胞癌（HCC），无论采用何种治疗方法，HCC 的复发率都很高。在这项研究中，我们试图提高效应 T 淋巴细胞对 HCC 的细胞溶解活性。单核细胞来源的树突状细胞（DC）通过用 HCC 细胞系的细胞裂解物或 RNA 冲击来激活 T 淋巴细胞。单核细胞通过用 IL4 和 GM-CSF 处理来激活分化为 DC。将 DC 用单个 HCC 细胞系或两个或三个 HCC 细胞系的组合制备的细胞裂解物或 RNA 冲击，然后与自体 T 淋巴细胞共培养，以产生特异性细胞毒性。我们发现，用总 RNA 冲击的 DC 比用总细胞裂解物冲击的 DC 更有效地发挥 T 淋巴细胞的功能，这表现在对 HCC 靶细胞的细胞溶解活性更高。在效应细胞：靶细胞比率为 10：1 时，Huh7、HepG2 和 SNU449 细胞的凋亡百分比分别为 42.6±4.5%（p=0.01）、33.6±3.1%（p=0.007）和 21.4±1.4%（p<0.001）。用来自三种细胞系的抗原混合物冲击的 DC 可使效应 T 淋巴细胞的细胞溶解功能提高约两倍（p<0.001），这表明这种方法应进一步开发并应用于 HCC 的过继转移治疗。
