Construction and Characterization of Adenovirus Vectors Encoding Aspartate-β-Hydroxylase to Preliminary Application in Immunotherapy of Hepatocellular Carcinoma
Dendritic cells (DCs) harboring tumor-associated antigen are supposed to be a potential immunotherapy for hepatocellular carcinoma (HCC). Aspartate-β-hydroxylase (AAH), an overexpressed tumor-associated cell surface protein, is considered as a promising biomarker and therapeutic target for HCC. In t...
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| Format: | Article |
| Language: | English |
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Wiley
2018-01-01
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| Series: | Journal of Immunology Research |
| Online Access: | http://dx.doi.org/10.1155/2018/9832467 |
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| author | Yujiao Zhou Feifei Liu Chengmin Li Guo Shi Xiaolei Xu Xue Luo Yuanling Zhang Jingjie Fu Aizhong Zeng Limin Chen |
| author_facet | Yujiao Zhou Feifei Liu Chengmin Li Guo Shi Xiaolei Xu Xue Luo Yuanling Zhang Jingjie Fu Aizhong Zeng Limin Chen |
| author_sort | Yujiao Zhou |
| collection | DOAJ |
| description | Dendritic cells (DCs) harboring tumor-associated antigen are supposed to be a potential immunotherapy for hepatocellular carcinoma (HCC). Aspartate-β-hydroxylase (AAH), an overexpressed tumor-associated cell surface protein, is considered as a promising biomarker and therapeutic target for HCC. In this study, we constructed adenovirus vector encoding AAH gene by gateway recombinant cloning technology and preliminarily explored the antitumor effects of DC vaccines harboring AAH. Firstly, the total AAH mRNA was extracted from human HCC tissues; the cDNA was amplified by RT-PCR, verified, and sequenced after TA cloning. Gateway technology was used and the obtained 18T-AAH was used as a substrate, to yield the final expression vector Ad-AAH-IRES2-EGFP. Secondly, bone marrow-derived DCs were infected by Ad-AAH-IRES2-EGFP to yield AAH-DC vaccines. Matured DCs were demonstrated by increased expression of CD11c, CD80, and MHC-II costimulatory molecules. A dramatically cell-killing effect of T lymphocytes coculturing with AAH-DCs on HepG2 HCC cell line was demonstrated by CCK-8 and FCM assays in vitro. More importantly, in an animal experiment, the lysis effect of cytotoxic T lymphocytes (CTLs) on HepG2 cells in the AAH-DC group was stronger than that in the control groups. In conclusion, the gateway recombinant cloning technology is a powerful method of constructing adenovirus vector, and the product Ad-AAH-IRES2-EGFP may present as a potential candidate for DC-based immunotherapy of HCC. |
| format | Article |
| id | doaj-art-a43c9f2f3a07479691a24f82532a829c |
| institution | Kabale University |
| issn | 2314-8861 2314-7156 |
| language | English |
| publishDate | 2018-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Immunology Research |
| spelling | doaj-art-a43c9f2f3a07479691a24f82532a829c2025-02-03T06:13:11ZengWileyJournal of Immunology Research2314-88612314-71562018-01-01201810.1155/2018/98324679832467Construction and Characterization of Adenovirus Vectors Encoding Aspartate-β-Hydroxylase to Preliminary Application in Immunotherapy of Hepatocellular CarcinomaYujiao Zhou0Feifei Liu1Chengmin Li2Guo Shi3Xiaolei Xu4Xue Luo5Yuanling Zhang6Jingjie Fu7Aizhong Zeng8Limin Chen9Department of Infectious Disease, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, ChinaDepartment of Infectious Disease, The People’s Hospital of Deyang City, Deyang, Sichuan Province 618000, ChinaDepartment of Infectious Disease, The Chongqing Fuling Center Hospital, Chongqing 400016, ChinaDepartment of General Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, ChinaDepartment of Infectious Disease, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, ChinaDepartment of Infectious Disease, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, ChinaDepartment of Infectious Disease, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, ChinaDepartment of Infectious Disease, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, ChinaDepartment of Infectious Disease, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, ChinaToronto General Research Institute, University of Toronto, Toronto, ON, CanadaDendritic cells (DCs) harboring tumor-associated antigen are supposed to be a potential immunotherapy for hepatocellular carcinoma (HCC). Aspartate-β-hydroxylase (AAH), an overexpressed tumor-associated cell surface protein, is considered as a promising biomarker and therapeutic target for HCC. In this study, we constructed adenovirus vector encoding AAH gene by gateway recombinant cloning technology and preliminarily explored the antitumor effects of DC vaccines harboring AAH. Firstly, the total AAH mRNA was extracted from human HCC tissues; the cDNA was amplified by RT-PCR, verified, and sequenced after TA cloning. Gateway technology was used and the obtained 18T-AAH was used as a substrate, to yield the final expression vector Ad-AAH-IRES2-EGFP. Secondly, bone marrow-derived DCs were infected by Ad-AAH-IRES2-EGFP to yield AAH-DC vaccines. Matured DCs were demonstrated by increased expression of CD11c, CD80, and MHC-II costimulatory molecules. A dramatically cell-killing effect of T lymphocytes coculturing with AAH-DCs on HepG2 HCC cell line was demonstrated by CCK-8 and FCM assays in vitro. More importantly, in an animal experiment, the lysis effect of cytotoxic T lymphocytes (CTLs) on HepG2 cells in the AAH-DC group was stronger than that in the control groups. In conclusion, the gateway recombinant cloning technology is a powerful method of constructing adenovirus vector, and the product Ad-AAH-IRES2-EGFP may present as a potential candidate for DC-based immunotherapy of HCC.http://dx.doi.org/10.1155/2018/9832467 |
| spellingShingle | Yujiao Zhou Feifei Liu Chengmin Li Guo Shi Xiaolei Xu Xue Luo Yuanling Zhang Jingjie Fu Aizhong Zeng Limin Chen Construction and Characterization of Adenovirus Vectors Encoding Aspartate-β-Hydroxylase to Preliminary Application in Immunotherapy of Hepatocellular Carcinoma Journal of Immunology Research |
| title | Construction and Characterization of Adenovirus Vectors Encoding Aspartate-β-Hydroxylase to Preliminary Application in Immunotherapy of Hepatocellular Carcinoma |
| title_full | Construction and Characterization of Adenovirus Vectors Encoding Aspartate-β-Hydroxylase to Preliminary Application in Immunotherapy of Hepatocellular Carcinoma |
| title_fullStr | Construction and Characterization of Adenovirus Vectors Encoding Aspartate-β-Hydroxylase to Preliminary Application in Immunotherapy of Hepatocellular Carcinoma |
| title_full_unstemmed | Construction and Characterization of Adenovirus Vectors Encoding Aspartate-β-Hydroxylase to Preliminary Application in Immunotherapy of Hepatocellular Carcinoma |
| title_short | Construction and Characterization of Adenovirus Vectors Encoding Aspartate-β-Hydroxylase to Preliminary Application in Immunotherapy of Hepatocellular Carcinoma |
| title_sort | construction and characterization of adenovirus vectors encoding aspartate β hydroxylase to preliminary application in immunotherapy of hepatocellular carcinoma |
| url | http://dx.doi.org/10.1155/2018/9832467 |
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