Zhao Jinjin, Zhang Haiguang, Zhao Yilin, Lin Zhiqiang, Lin Fei, Wang Zhiyin, Mo Qingjiang, Lu Guangjian, Zhao Guoan, Wang Guoqiang
Clinical Laboratory, the First Affiliated Hospital of Xinxiang Medical University, Xinxiang, People's Republic of China.
Key Laboratory of Nano-Drug Delivery System Construction and Application in Xinxiang City, the First Affiliated Hospital of Xinxiang Medical University, Xinxiang, People's Republic of China.
Cancer Manag Res. 2024 Dec 17;16:1813-1822. doi: 10.2147/CMAR.S482363. eCollection 2024.
Tumor-specific antigens play an important role in dendritic cell (DC)-based immunotherapy. The acquisition of tumor-specific antigens, which are essential for DC-based immunotherapy, poses a significant challenge. This study aimed to explore the efficacy of hypoxia inducible factor-1α (HIF-1α) overexpression tumor antigens in DC-based immunotherapy.
An HIF-1α over-expression cell line was constructed to prepare HIF-1α overexpression tumor antigens. The expression of CD14, CD40, CD80, CD86, and HLA-DR on the surface of dendritic cells derived from monocytes was assessed using flow cytometry after stimulation with tumor antigens enriched in HIF-1α. T cell proliferation was analyzed by CFSE division following incubation with mature DCs. The apoptotic tumor cells were detected through annexin V/PI staining following coculture with dendritic cells (DCs) stimulated by HIF-1α enriched antigens. The detection of damage-associated molecular pattern molecules (DAMPs) HMGB1 and calreticulin (CALR) was performed using Western blotting.
The results demonstrated that HIF-1α-enriched tumor antigens significantly upregulated the expression of CD40, CD80, CD86, and HLA-DR in DCs compared to normal tumor antigens. Furthermore, co-incubation with HIF-1α-enriched tumor antigen-activated DCs enhanced T cell proliferation and stimulated the T cell-mediated cytotoxicity. Notably, the expression of DAMPs, such as HMGB1 and CALR, was elevated in HIF-1α-enriched tumor antigens.
Our findings demonstrate that tumor antigens enriched with HIF-1α may encompass tumor-specific antigens capable of stimulating DC activation, thereby enhancing T cell proliferation and cytotoxicity. These results provide support for the further advancement of HIF-1α enriched tumor antigens in preclinical and clinical investigations pertaining to tumor treatment.
肿瘤特异性抗原在基于树突状细胞（DC）的免疫治疗中发挥着重要作用。获取肿瘤特异性抗原是基于DC的免疫治疗所必需的，但这构成了一项重大挑战。本研究旨在探讨缺氧诱导因子-1α（HIF-1α）过表达肿瘤抗原在基于DC的免疫治疗中的疗效。
构建HIF-1α过表达细胞系以制备HIF-1α过表达肿瘤抗原。用富含HIF-1α的肿瘤抗原刺激后，采用流式细胞术评估源自单核细胞的树突状细胞表面CD14、CD40、CD80、CD86和HLA-DR的表达。与成熟DC孵育后，通过CFSE分裂分析T细胞增殖。与经富含HIF-1α抗原刺激的树突状细胞（DC）共培养后，通过膜联蛋白V/PI染色检测凋亡肿瘤细胞。使用蛋白质免疫印迹法检测损伤相关分子模式分子（DAMPs）高迁移率族蛋白B1（HMGB1）和钙网蛋白（CALR）。
结果表明，与正常肿瘤抗原相比,富含HIF-1α的肿瘤抗原显著上调DC中CD40、CD80、CD86和HLA-DR的表达。此外，与富含HIF-1α的肿瘤抗原激活的DC共同孵育可增强T细胞增殖并刺激T细胞介导的细胞毒性。值得注意的是，在富含HIF-1α的肿瘤抗原中，DAMPs如HMGB1和CALR的表达升高。
我们的研究结果表明，富含HIF-1α的肿瘤抗原可能包含能够刺激DC激活的肿瘤特异性抗原，从而增强T细胞增殖和细胞毒性。这些结果为富含HIF-1α的肿瘤抗原在肿瘤治疗的临床前和临床研究中的进一步发展提供了支持。
