title: Fusion with human lung cancer cells elongates the life span of human umbilical endothelial cells and enhances the anti-tumor immunity.
abstract: OBJECTIVE
  Human umbilical endothelial cells (HUVECs) have been proved as an effective whole-cell vaccine inhibiting tumor angiogenesis. However, HUVECs divide a very limited number of passages before entering replicative senescence, which limits its application for clinical situation. Here, we fused HUVECs with human pulmonary adenocarcinoma cell line A549s and investigated the anti-tumor immunity of the hybrids against mice Lewis lung cancer.
  
  
  METHODS
  HUVECs were fused with A549s using polyethylene glycol and were sorted by flow cytometry. The fusion cells (HUVEC-A549s) were confirmed by testing the expression of telomerase and VE-cadherin, the senescence-associated β-galactosidase activity, and tube formation ability. HUVEC-A549s were then irradiated and injected into the C57BL/6 mice of protective, therapeutic, and metastatic models. The mechanism of the anti-tumor immunity was explored by analyzing mice sera, spleen T lymphocytes, tumor microenvironment, and histological changes.
  
  
  RESULTS
  HUVEC-A549s coexpressed tumor and endothelial markers and maintained the vascular function of tube forming at passage 30 without showing signs of senescence. HUVEC-A549s could induce protective and therapeutic anti-tumor activity for LL(2) model and presented stronger activity against metastasis than HUVECs. Both humoral and cellular immunity were participated in the anti-angiogenic activity, as HUVECs-neutralizing IgG and HUVECs-toxic lymphocytes were increased. Angiogenic mediators (VEGF and TGF-β) and tumor microenvironment cells MDSCs and Tregs were also diminished.
  
  
  CONCLUSIONS
  Our findings might provide a novel strategy for HUVECs-related immunotherapy, and this vaccine requires lower culture condition than primary HUVECs while enhancing the anti-tumor immunity.
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University, Chengdu, 610041, Sichuan, People's Republic of China.|Department of Gynecology and Obstetrics, West China Second Hospital, Sichuan University, Chengdu, 610041, Sichuan, People's Republic of China.
authors.forename: Xiyan
authors.initials: X
authors.lastname: Mu
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University, Chengdu, 610041, Sichuan, People's Republic of China.
authors.forename: Chunju
authors.initials: C
authors.lastname: Fang
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University, Chengdu, 610041, Sichuan, People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University, Chengdu, 610041, Sichuan, People's Republic of China.
authors.forename: Yufeng
authors.initials: Y
authors.lastname: Xi
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University, Chengdu, 610041, Sichuan, People's Republic of China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University, Chengdu, 610041, Sichuan, People's Republic of China.
authors.forename: Yuquan
authors.initials: Y
authors.lastname: Wei
authors.affiliation: Department of Gynecology and Obstetrics, West China Second Hospital, Sichuan University, Chengdu, 610041, Sichuan, People's Republic of China.
authors.forename: Tao
authors.initials: T
authors.lastname: Yi
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital and Collaborative Innovation Center for Biotherapy, Sichuan University, Chengdu, 610041, Sichuan, People's Republic of China. stillwy@163.com.
authors.forename: Yang
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Department of Gynecology and Obstetrics, West China Second Hospital, Sichuan University, Chengdu, 610041, Sichuan, People's Republic of China. drzhaoxia@163.com.
authors.forename: Xia
authors.initials: X
authors.lastname: Zhao
mesh_terms: D000818:Animals; D015153:Blotting, Western; D018827:Carcinoma, Lewis Lung / Q000098:blood supply / Q000276:immunology / Q000517:prevention & control*; D049109:Cell Proliferation; D002478:Cells, Cultured; D016922:Cellular Senescence; D004730:Endothelium, Vascular / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D061307:Human Umbilical Vein Endothelial Cells / Q000276:immunology; D006801:Humans; D007111:Immunity, Cellular / Q000276:immunology*; D007124:Immunoenzyme Techniques; D051379:Mice; D008810:Mice, Inbred C57BL; D009389:Neovascularization, Pathologic / Q000276:immunology*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D016212:Transforming Growth Factor beta / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology / Q000276:immunology*; D014611:Vaccination
chemical_list: D012333:RNA, Messenger; D016212:Transforming Growth Factor beta
keywords: A549; Cancer biotherapy; HUVEC; Hybrid; Vaccine