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表达CCL19的溶瘤腺病毒增强小鼠对胃癌的免疫力

[Oncolytic adenovirus expressing CCL19 enhances immunity against gastric cancer in mice].

作者信息

Chen Zheng, Liu Jiheng, Xu Dayong

机构信息

Department of General Surgery, Changsha First Hospital, Changsha 410005, China.

Department of General Surgery, Changsha First Hospital, Changsha 410005, China. *Corresponding author, E-mail:

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2021 Feb;37(2):119-124.

Abstract

Objective To explore whether oncolytic adenovirus expressing CCL19 can inhibit the growth of gastric cancer cells and activate anti-tumor immune response. Methods Mouse CCL19 gene was inserted into the E3 region of oncolytic adenovirus Ad5 to obtain engineered oncolytic adenovirus Ad5-CCL19. The expression of CCL19 in Ad5-CCL19-infected mouse MFC cells was detected by Western blotting. The effects of Ad5-CCL19 on the proliferation of MFC cells, MGC803 cells and BGC823 cells were tested by MTT assay. The anti-tumor activity of Ad5-CCL19 in vivo was examined by MFC cell subcutaneous transplantation tumor model. Immunofluorescence histochemical staining was used to detect CD4 and CD8 expression in tumor tissue. The secretion levels of interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) in tumor infiltrating T cells were detected by flow cytometry. Results Ad5-CCL19 could effectively infect MFC cells to secrete CCL19. Also, Ad5-CCL19 could induce significant dose-dependent cytotoxicity against target cells in vitro. The experiment in vivo showed that Ad5-CCL19 had stronger inhibitory effects on MFC cell tumor than Ad5 in the mice, and it could effectively enhance the infiltration of CD4 T cells and CD8 T cells and increase the secretion of IFN-γ and TNF-α in tumor tissues. Conclusion Ad5-CCL19 can significantly infect MFC gastric cancer cells to inhibit their growth and improve the anti-tumor immune activity of the tumor site.

摘要

目的 探讨表达CCL19的溶瘤腺病毒是否能抑制胃癌细胞生长并激活抗肿瘤免疫反应。方法 将小鼠CCL19基因插入溶瘤腺病毒Ad5的E3区,获得工程化溶瘤腺病毒Ad5-CCL19。通过蛋白质免疫印迹法检测Ad5-CCL19感染的小鼠MFC细胞中CCL19的表达。采用MTT法检测Ad5-CCL19对MFC细胞、MGC803细胞和BGC823细胞增殖的影响。利用MFC细胞皮下移植瘤模型检测Ad5-CCL19在体内的抗肿瘤活性。采用免疫荧光组织化学染色法检测肿瘤组织中CD4和CD8的表达。通过流式细胞术检测肿瘤浸润T细胞中干扰素-γ(IFN-γ)和肿瘤坏死因子-α(TNF-α)的分泌水平。结果 Ad5-CCL19能有效感染MFC细胞并分泌CCL19。此外,Ad5-CCL19在体外能诱导对靶细胞显著的剂量依赖性细胞毒性。体内实验表明,Ad5-CCL19对小鼠MFC细胞肿瘤的抑制作用比Ad5更强,且能有效增强CD4 T细胞和CD8 T细胞的浸润,并增加肿瘤组织中IFN-γ和TNF-α的分泌。结论 Ad5-CCL19能显著感染MFC胃癌细胞,抑制其生长,并提高肿瘤部位的抗肿瘤免疫活性。

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