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携带人白细胞介素2的重组溶瘤痘苗病毒的抗肿瘤活性

Antitumor activity of recombinant oncolytic vaccinia virus with human IL2.

作者信息

Liu Liqiong, Li Huiqun, Xu Qinggang, Wu Yan, Chen Dongfeng, Yu Feng

机构信息

Department of Hematology, Huazhong University of Science and Technology Union Shenzhen Hospital, Shenzhen, Guangdong Province, 518052, P. R. China.

School of Life Sciences, Jiangsu University, Zhenjiang, 212013, P. R. China.

出版信息

Open Med (Wars). 2022 Jun 17;17(1):1084-1091. doi: 10.1515/med-2022-0496. eCollection 2022.

Abstract

The tumor microenvironment is highly immunosuppressive. The genetically modified oncolytic vaccinia virus (OVV) is a promising vector for cancer immunotherapy. The aim of the present study was to assess the antitumor effects of human interleukin-2 (hIL2)-armed OVV . The hIL2 gene was inserted into a thymidine kinase and the viral growth factor double deleted oncolytic VV (VVDD) to generate recombinant hIL2-armed OVV (rVVDD-hIL2). Viral replication capacity in A549 cells was quantified by plaque titration on CV-1 cells. Production of hIL2 in cancer cells infected by rVVDD-hIL2 was measured by enzyme-linked immunosorbent assay. Finally, 3-(4,5-dimethylthiazol-2-yl)-5-(3-arboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay was performed to assess the antitumor effects of rVVDD-hIL2. The results showed that rVVDD-hIL2 viral particles expressed increasing levels of hIL2 in human and murine cancer cell lines with growing multiplicities of infection (MOIs). The insertion of the hIL2 gene did not impair the replication capacity of VV, and the rVVDD-hIL2 virus killed cancer cells efficaciously. The lytic effects of the recombinant oncolytic virus on tumor cells increased with the growing MOIs. In conclusion, these findings suggest that hIL2-armed VVDD effectively infects and lyses tumor cells, with high expression of hIL2.

摘要

肿瘤微环境具有高度免疫抑制性。基因改造的溶瘤痘苗病毒(OVV)是一种很有前景的癌症免疫治疗载体。本研究的目的是评估携带人白细胞介素-2(hIL2)的OVV的抗肿瘤作用。将hIL2基因插入胸苷激酶和病毒生长因子双缺失的溶瘤痘苗病毒(VVDD)中,以产生重组携带hIL2的OVV(rVVDD-hIL2)。通过在CV-1细胞上进行噬斑滴定来定量A549细胞中的病毒复制能力。用酶联免疫吸附测定法测量受rVVDD-hIL2感染的癌细胞中hIL2的产生。最后,进行3-(4,5-二甲基噻唑-2-基)-5-(3-羧基甲氧基苯基)-2-(4-磺基苯基)-2H-四唑内盐(MTS)测定,以评估rVVDD-hIL2的抗肿瘤作用。结果表明,随着感染复数(MOIs)的增加,rVVDD-hIL2病毒颗粒在人和鼠癌细胞系中表达的hIL2水平不断升高。hIL2基因的插入并未损害痘苗病毒的复制能力,并且rVVDD-hIL2病毒能有效杀死癌细胞。重组溶瘤病毒对肿瘤细胞的裂解作用随着MOIs的增加而增强。总之,这些发现表明携带hIL2的VVDD能有效感染并裂解肿瘤细胞,且hIL2表达水平高。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8ee/9206501/148759a18039/j_med-2022-0496-fig001.jpg

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