title: CCL21/IL21-armed oncolytic adenovirus enhances antitumor activity against TERT-positive tumor cells.
abstract: Multigene-armed oncolytic adenoviruses are capable of efficiently generating a productive antitumor immune response. The chemokine (C-C motif) ligand 21 (CCL21) binds to CCR7 on naïve T cells and dendritic cells (DCs) to promote their chemoattraction to the tumor and resultant antitumor activity. Interleukin 21 (IL21) promotes survival of naïve T cells while maintaining their CCR7 surface expression, which increases their capacity to transmigrate in response to CCL21 chemoattraction. IL21 is also involved in NK cell differentiation and B cell activation and proliferation. The generation of effective antitumor immune responses is a complex process dependent upon coordinated interactions of various subsets of effector cells. Using the AdEasy system, we aimed to construct an oncolytic adenovirus co-expressing CCL21 and IL21 that could selectively replicate in TERTp-positive tumor cells (Ad-CCL21-IL21 virus). The E1A promoter of these oncolytic adenoviruses was replaced by telomerase reverse transcriptase promoter (TERTp). Ad-CCL21-IL21 was constructed from three plasmids, pGTE-IL21, pShuttle-CMV-CCL21 and AdEasy-1 and was homologously recombined and propagated in the Escherichia coli strain BJ5183 and the packaging cell line HEK-293, respectively. Our results showed that our targeted and armed oncolytic adenoviruses Ad-CCL21-IL21 can induce apoptosis in TERTp-positive tumor cells to give rise to viral propagation, in a dose-dependent manner. Importantly, we confirm that these modified oncolytic adenoviruses do not replicate efficiently in normal cells even under high viral loads. Additionally, we investigate the role of Ad-CCL21-IL21 in inducing antitumor activity and tumor specific cytotoxicity of CTLs in vitro. This study suggests that Ad-CCL21-IL21 is a promising targeted tumor-specific oncolytic adenovirus.
authors.affiliation: Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, China-Japan Union Hospital, Jilin University, Changchun 130062, PR China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Li
authors.affiliation: The Affiliated Hospital to Changchun University of Chinese Medicine, Changchun 130021, PR China.
authors.forename: Yi-Fei
authors.initials: YF
authors.lastname: Li
authors.affiliation: Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, China-Japan Union Hospital, Jilin University, Changchun 130062, PR China.
authors.forename: Chong-Zhan
authors.initials: CZ
authors.lastname: Si
authors.affiliation: Second Hospital of Jilin University, Changchun, Jilin Province 130021, PR China.
authors.forename: Yu-Hui
authors.initials: YH
authors.lastname: Zhu
authors.affiliation: Second Hospital of Jilin University, Changchun, Jilin Province 130021, PR China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Jin
authors.affiliation: Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, China-Japan Union Hospital, Jilin University, Changchun 130062, PR China.
authors.forename: Tong-Tong
authors.initials: TT
authors.lastname: Zhu
authors.affiliation: Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, China-Japan Union Hospital, Jilin University, Changchun 130062, PR China; Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, PR China.
authors.forename: Ming-Yuan
authors.initials: MY
authors.lastname: Liu
authors.affiliation: Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, China-Japan Union Hospital, Jilin University, Changchun 130062, PR China. Electronic address: myth03171@yeah.net.
authors.forename: Guang-Yao
authors.initials: GY
authors.lastname: Liu
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000276:immunology; D017500:Adenovirus E1A Proteins / Q000235:genetics* / Q000276:immunology; D018938:Caco-2 Cells; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D054421:Chemokine CCL21 / Q000235:genetics* / Q000276:immunology; D018920:Coculture Techniques; D004847:Epithelial Cells / Q000166:cytology / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D019073:HT29 Cells; D006367:HeLa Cells; D059765:Homologous Recombination; D006801:Humans; D007378:Interleukins / Q000235:genetics* / Q000276:immunology; D008297:Male; D050130:Oncolytic Virotherapy / Q000379:methods; D050504:Oncolytic Viruses / Q000235:genetics* / Q000276:immunology; D010957:Plasmids / Q000737:chemistry / Q000276:immunology; D011467:Prostate / Q000166:cytology / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology / Q000276:immunology; D000074084:THP-1 Cells; D019098:Telomerase / Q000235:genetics* / Q000276:immunology; D000098546:Interleukin-21
chemical_list: D017500:Adenovirus E1A Proteins; C515478:CCL21 protein, human; D054421:Chemokine CCL21; D007378:Interleukins; C509186:TERT protein, human; D019098:Telomerase; D000098546:Interleukin-21
keywords: CCL21; Combination gene therapy; IL21; Oncolytic adenoviruses; Tissue-specific promoter