Exploratory Research for HIF-1α Overexpression Tumor Antigen in the Activation of Dendritic Cells and the Potent Anti-Tumor Immune Response

Jinjin Zhao,1,2 Haiguang Zhang,3 Yilin Zhao,4 Zhiqiang Lin,1 Fei Lin,5,6 Zhiyin Wang,1 Qingjiang Mo,1 Guangjian Lu,1 Guoan Zhao,5,6 Guoqiang Wang1 1Clinical Laboratory, the First Affiliated Hospital of Xinxiang Medical University, Xinxiang, People’s Republic of China; 2Key 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; 3Department of Gynecology, the First Affiliated Hospital of Xinxiang Medical University, Xinxiang, People’s Republic of China; 4Department of Cardiology, the Third Affiliated Hospital of Xinxiang Medical University, Xinxiang, People’s Republic of China; 5Department of Cardiology, the First Affiliated Hospital of Xinxiang Medical University, Xinxiang, People’s Republic of China; 6Cardiovascular Repair Engineering Technology Research Center, the First Affiliated Hospital of Xinxiang Medical University, Xinxiang, People’s Republic of ChinaCorrespondence: Guoan Zhao; Guoqiang Wang, Email [email protected]; [email protected]: 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.Methods: 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.Results: 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.Conclusion: 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.Keywords: dendritic cells, T cell activation, HIF-1α, HMGB1, calreticulin