Pan Z, Li S, Wang Y, Liu H, Gui L, Dong B
Department of Medical Microbiology and Immunology,Wannan Medical College, Wuhu 241002, China.
Department of Biochemistry,Wannan Medical College, Wuhu 241002, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2023 Jun 20;43(6):906-914. doi: 10.12122/j.issn.1673-4254.2023.06.05.
To assess the effect of tumor cell lysate (TCL) with low high-mobility group B1 (HMGB1) content for enhancing immune responses of dendritic cells (DCs) against lung cancer.
TCLs with low HMGB1 content (LH-TCL) and normal HMGB1 content (NH-TCL) were prepared using Lewis lung cancer (LLC) cells in which HMGB1 was inhibited with 30 nmol/L glycyrrhizic acid (GA) and using LLC cells without GA treatment, respectively. Cultured mouse DCs were exposed to different doses of NH-TCL and LH-TCL, using PBS as the control. Flow cytometry was used to detect the expressions of CD11b, CD11c and CD86 and apoptosis of the stimulated DCs, and IL-12 levels in the cell cultures were detected by ELISA. Mouse spleen cells were co-cultured with the stimulated DCs, and the activation of the spleen cells was assessed by detecting CD69 expression using flow cytometry; TNF-β production in the spleen cells was detected with ELISA. The spleen cells were then co-cultured with LLC cells at the effector: target ratios of 5:1, 10:1 and 20:1 to observe the tumor cell killing. In the animal experiment, C57/BL6 mouse models bearing subcutaneous LLC xenograft received multiple injections with the stimulated DCs, and the tumor growth was observed.
The content of HMGB1 in the TCL prepared using GA-treated LLC cells was significantly reduced ( < 0.01). Compared with NH-TCL, LH-TCL showed a stronger ability to reduce apoptosis ( < 0.001) and promote activation and IL- 12 production in the DCs. Compared with those with NH-TCL stimulation, the DCs stimulated with LH-TCL more effectively induced activation of splenic lymphocytes and enhanced their anti-tumor immunity ( < 0.05). In the cell co-cultures, the spleen lymphocytes activated by LH-TCL-stimulated DCs showed significantly enhanced LLC cell killing activity ( < 0.01). In the tumor-bearing mice, injections of LH-TCL-stimulated DCs effectively activated host anti-tumor immunity and inhibited the growth of the tumor xenografts ( < 0.05).
Stimulation of the DCs with LH-TCL enhances the anti-tumor immune activity of the DCs and improve the efficacy of DCbased immunotherapy for LLC in mice.
评估高迁移率族蛋白B1（HMGB1）含量低的肿瘤细胞裂解物（TCL）增强树突状细胞（DCs）对肺癌免疫反应的效果。
分别使用经30 nmol/L甘草酸（GA）抑制HMGB1的Lewis肺癌（LLC）细胞和未经GA处理的LLC细胞制备HMGB1含量低的TCL（LH-TCL）和正常HMGB1含量的TCL（NH-TCL）。将培养的小鼠DCs暴露于不同剂量的NH-TCL和LH-TCL，以PBS作为对照。采用流式细胞术检测受刺激DCs的CD11b、CD11c和CD86表达及凋亡情况，通过ELISA检测细胞培养物中的IL-12水平。将小鼠脾细胞与受刺激的DCs共培养，通过流式细胞术检测CD69表达评估脾细胞的活化情况；用ELISA检测脾细胞中TNF-β的产生。然后将脾细胞与LLC细胞按效应细胞：靶细胞比例为5:1、10:1和20:1共培养，观察肿瘤细胞杀伤情况。在动物实验中，对皮下接种LLC异种移植物的C57/BL6小鼠模型多次注射受刺激的DCs，观察肿瘤生长情况。
使用经GA处理的LLC细胞制备的TCL中HMGB1含量显著降低（<0.01）。与NH-TCL相比，LH-TCL降低DCs凋亡的能力更强（<0.001），并能促进DCs的活化和IL-12产生。与经NH-TCL刺激的DCs相比，经LH-TCL刺激的DCs更有效地诱导脾淋巴细胞活化并增强其抗肿瘤免疫力（<0.05）。在细胞共培养中，经LH-TCL刺激的DCs活化的脾淋巴细胞对LLC细胞的杀伤活性显著增强（<0.01）。在荷瘤小鼠中，注射经LH-TCL刺激的DCs可有效激活宿主抗肿瘤免疫力并抑制肿瘤异种移植物的生长（<0.05）。
用LH-TCL刺激DCs可增强DCs的抗肿瘤免疫活性，提高小鼠LLC基于DC的免疫治疗效果。
