title: Identification of Optimal Insertion Site in Recombinant Newcastle Disease Virus (rNDV) Vector Expressing Foreign Gene to Enhance Its Anti-Tumor Effect.
abstract: Recombinant Newcastle disease virus (rNDV) is tumor selective and intrinsically oncolytic, which has been developed as a vector to express exogenous genes to enhance its oncolytic efficacy. Our previous studies found that insertion sites of foreign gene in rNDV vector affected its expression and anti-tumor activities. However, the optimal insertion site for foreign genes remains unknown. In this study, we inserted the enhanced green fluorescence protein (EGFP) and IL2 genes into four different intergenic regions of the rNDV using reverse genetics technology. Recombinants rNDV-EGFPs and rNDV-IL2s were successfully rescued, which displayed the similar growth kinetics with parental virus. Both EGFP mRNA and protein levels were most abundant in HepG2 cells, when EGFP gene was inserted between the NP/P site of the rNDV. Similarly, the IL-2 expressed by HepG2 cells infected with rNDV-IL2 was highest, when IL2 was inserted into NP/P site. To test whether these rNDVs that express higher foreign genes could induce stronger anti-tumor response, we treated the H22-oxter-tumor-bearing C57BL/6J mice with rNDV-IL2s and then examined the oncolytic efficacy. The results showed that rNDV-IL2-NP/P had the strongest inhibition of murine hepatoma carcinoma tumors. The splenocytes isolated from the mice treated with rNDV-IL2-NP/P reached the highest degrees of CD4+ T and CD8+ T cells. In addition, animals' survival rate in rNDV-IL2-NP/P-treated group was higher than that of other groups. Taken together, these results demonstrate that NP and P gene junction in rNDV is the optimal insertion site for foreign genes expression to enhance rNDV's anti-tumor effects.
authors.affiliation: College of Life Science and Technology, HeiLongJiang BaYi Agricultural University, Daqing, 163319, China.|College of Life Science, Northeast Agricultural University, Harbin, 150030, China.
authors.forename: Ziye
authors.initials: Z
authors.lastname: Pan
authors.affiliation: College of Life Science, Northeast Agricultural University, Harbin, 150030, China.
authors.forename: Jinjiao
authors.initials: J
authors.lastname: He
authors.affiliation: College of Life Science, Northeast Agricultural University, Harbin, 150030, China.
authors.forename: Lubna M
authors.initials: LM
authors.lastname: Rasoul
authors.affiliation: College of Life Science, Northeast Agricultural University, Harbin, 150030, China.
authors.forename: Yunye
authors.initials: Y
authors.lastname: Liu
authors.affiliation: College of Life Science, Northeast Agricultural University, Harbin, 150030, China.
authors.forename: Ruixiang
authors.initials: R
authors.lastname: Che
authors.affiliation: College of Life Science and Technology, HeiLongJiang BaYi Agricultural University, Daqing, 163319, China.|College of Life Science, Northeast Agricultural University, Harbin, 150030, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Ding
authors.affiliation: College of Life Science, Northeast Agricultural University, Harbin, 150030, China.
authors.forename: Xiaocheng
authors.initials: X
authors.lastname: Guo
authors.affiliation: College of Life Science, Northeast Agricultural University, Harbin, 150030, China.
authors.forename: Jiarui
authors.initials: J
authors.lastname: Yang
authors.affiliation: College of Life Science and Technology, HeiLongJiang BaYi Agricultural University, Daqing, 163319, China.|College of Life Science, Northeast Agricultural University, Harbin, 150030, China.
authors.forename: Dehua
authors.initials: D
authors.lastname: Zou
authors.affiliation: College of Life Science and Technology, HeiLongJiang BaYi Agricultural University, Daqing, 163319, China.
authors.forename: Hua
authors.initials: H
authors.lastname: Zhang
authors.affiliation: College of Life Science, Northeast Agricultural University, Harbin, 150030, China.
authors.forename: Deshan
authors.initials: D
authors.lastname: Li
authors.affiliation: College of Life Science and Technology, HeiLongJiang BaYi Agricultural University, Daqing, 163319, China.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Cao
mesh_terms: D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology; D005260:Female; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D056945:Hep G2 Cells; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics / Q000378:metabolism; D008113:Liver Neoplasms / Q000401:mortality / Q000473:pathology / Q000628:therapy / Q000662:veterinary; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D009522:Newcastle disease virus / Q000235:genetics*; D015996:Survival Rate; D014184:Transplantation, Homologous
chemical_list: D007376:Interleukin-2; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins